Aggressive angiomyxoma is a rare benign mesenchymal tumor and occurs rarely in males. This study aimed to review all the cases of AAM in men in the English literature up to September 2020 and investigate the clinical, histochemical, and radiological characteristics of AAM and discuss the best treatment choices according to available data.A comprehensive search of the PubMed, Google Scholar, and Embase databases up to September 2020 was performed looking for reported cases of male patients with AAM. The search excluded articles in languages other than English, reported female cases, and superficial angiomyxoma cases.Among the 97 patients, the mean age was 48.2 years with an incidence peak between 40 and 60 years. The sites commonly involved were the scrotum (42.3%). On ultrasound, the tumor was hypoechoic (85.7%) with a well-defined margin (100%), whereas on MRI, most cases were isointense on T1-weighted images (53.8%), and hyperintense on T2-weighted images (85.7%). Immunohistochemistry revealed that the tumor tended to be positive for vimentin (100%), CD34 (63.4%), ER (50%), and PR (53.3%) while S-100 showed 91% negativity. Wide and complete surgical excision was conducted in most cases (72%), and follow-up duration ranged from 1 month to 144 months with a recurrence rate of 11.8%.Although the occurrence of AAM is rare in men, consideration should be taken in the differential diagnosis of a mass in the genitourinary region. According to our review, the most decisive immunohistochemistry profile is the positivity of Vimentin and CD34 with the negativity of S-100. Although hormonal treatment is controversial, we suggest a novel algorithm for the management of aggressive angiomyxoma. •Aggressive angiomyxoma is a rare benign mesenchymal tumor that occurs rarely in men compared to the woman.•Less than 90 cases in men have been reported up to September 2020.•Radiologic imaging play role in the initial approach and the follow-up.•The most decisive immunohistochemistry profile is the positivity of Vimentin and CD34 with the negativity of S-100.•This review presents a novel algorithm for the management of aggressive angiomyxoma. It occurs commonly in females at the productive age with a male to female ratio of 1:6 . The firHere we report on a rare clinical case of aggressive angiomyxoma in man and review systemically all cases in men up to September 2020.2A 72-year-old man presented to our out-patient clinic at Aleppo university hospital complaining of an increase in the size of the right testicle without pain, redness, or localized heat. Regarding his past medical history, he has had hypertension for 20 years, benign prostatic hypertrophy for 20 years, chronic hemorrhoids, and chronic laxative-dependent constipation. He has no past surgical history. Drugs history included aspirin 100 mg, Proton-pump inhibitors (PPIs) over ten years, Tamsulosin, herbal laxatives for 25 years, and neurotoxic medications Luxstan 1.5 mg. The patient is a smoker, smoking approximately 58 packets/year, and consumes alcohol approximately once a week for 50 years. The patient has no allergies or traumatic histories, and his family history was clear of malignancies. On clinical examination, there was palpation of a hard, moving, painless mass above the right testicle.Laboratory tests showed a mildly raised level of serum glucose, but other biochemical and hormonal tests were normal. Beta-human chorionic gonadotropins (B-HCG), alpha-fetoprotein, and prostate-specific antigen (PSA) markers were also within normal limits. A scrotal ultrasound showed a well-defined, oval, hypoechoic, homogeneous tissue above the right testis measuring 6.1 × 4.5 cm with internal vascularity. It squeezes the medial part of the right epididymis head . Our surMacroscopically, a testis measuring 5 × 6.5 × 9 with a spermatic cord. Cut section showed para-testicular well-defined mass measuring 6.5 cm with a greater diameter. Microscopically, a poorly circumscribed infiltrative tumor made up of bland, spindled, and stellate cells with delicate cytoplasmic processes that surrounded blood vessels, on the myxoid stroma. There were no atypia or mitotic figures. Immunohistochemically, tumor cells were positive for estrogen receptors (ER), progesterone receptors (PR), and focally for smooth muscle actin (SMA), while negative for calretinin, CD34, desmin, and S-100 protein. Additionally, Ki67 proliferation marker was low (<5%) . The finMultislice computed tomography (CT) scan for chest, abdomen, and pelvic was performed ten days after surgery and showed no metastases, lymphadenopathy, or free fluids. Two years postoperatively, the patient was fine without signs of recurrence, as determined by magnetic resonance imaging (MRI) and CT scan.33.1: 10.17605/OSF.IO/U64WB. In addition, the case report has been prepared and reported in line with the SCARE guidelines [A literature search of computerized medical literature was performed using the PubMed, EMBASE, and Google Scholar. The keywords searched were (“angiomyxoma” OR “aggressive angiomyxoma”) AND . The search terms were modified to fit each database. The search was conducted for published papers up to September 2020, and restricted to the English language. To ensure that the search was complete, reference lists and similar articles were manually searched to identify additional relevant studies. We conducted our systematic review following to recommendations of Preferred Reporting Items for Systematic Reviews and Meta-Analyses 2020 (PRISMA 2020)  and it i3.2Studies were included of cases involved angiomyxoma in male adults and children in the reproductive tract, pelvis, perineum, or extra-genitourinary region. Additionally, if the type of case report, case series, or any literature review contained a case report the study was included. Articles that had female cases, were in languages other than English, or involved superficial angiomyoxoma were excluded.3.3.Screening and data extraction was done by two review authors (AYA and BT) using the Rayyan tool for screening and Microsoft Excel 2010 for data extraction, and was checked by a third author (KhA) to resolve any disagreement. Data was analyzed using IBM SPSS Statistics 244The first case in men was reported by Bégin et al., in 1985. Up to September 2020, 88 cases in men were reported with two peaks of publishing in 2008 and 2017. Among the 97 patients, the age of presentation varied from 9 months to 81 years  with incidence peak between 40 and 60 years. The genitourinary region was commonly involved (85.6%), and the scrotum was the most common region compare to the extra-genitourinary region (14.4%) . The maxThe tumor was hypoechoic on US  with well-defined margins  and no difference in internal echogenicity. On MRI, the most cases were isointense on T1-weighted images  and hyperintense on T2-weighted images . Regarding immunohistochemistry, the tumor tended to be positive for vimentin , CD34 , ER , and PR , while S-100 showed 91% negativity (n = 41/45). Alcian blue, Factor VII, and androgen receptor were 100% positive but present in the minority of cases. Fifteen cases out of sixteen were positive for Ki67 receptors with less than 10% and only one case out of sixteen had a 15% positivity percentage of the cells .Table 2SThe surgical approach varied, wide and complete surgical excision was conducted in the most cases , while frequency of local surgical excision and excisional biopsy was . Four patients were treated with hormonal treatment, and in two of them, the tumor was positive for estrogen and progesterone receptors. Radiotherapy was conducted in 2 patients and chemotherapy in 1 patient. Recurrence rate was 11.8% (n = 8/68) and the mean time period for recurrence was 28.8 months. The minimum time period until recurrence was 2 months, while the maximum time period was 84 months. Follow-up duration ranged from 1 months to 144 months .5AAM is a benign mesenchymal tumor that arises mostly from the soft tissues of the pelvic region in premenopausal women, which was first reported in 1983 by Steeper and Rosai, and 2 years later, the first case in a man was described by Bégin. AAM shows female predominance, as less than 90 male cases have been reported to date ,8. In meImaging studies play an important role to make the diagnosis preoperatively when the malignancy is suspected. On US, the tumor revealed a hypoechoic, heterogeneous mass with a well-defined margin. However, one case showed a hyperechoic mass. MRI provides more advantages in identifying the characteristics of such mesenchymal tumors, including isointense or hypointense characteristics on T1-weighted images and hyperintense characteristics on T2-weighted images. One case showed hypointensity on the T2-weighted image and another iso-hyperintensity. The swirled sign observed on both US and MRI has also been mentioned in a recent study . TypicalHistologically, aggressive angiomyxomas consist of mesenchymal spindle-shaped or stellate cells embedded in a myxoid matrix with abundant fibroblasts, myofibroblasts, and variably sized vessels. Angiomyxomas show low mitotic activity with an absence of nuclear atypia , which cWide surgical excision is the treatment of choice, including broad free margins around the tumor in the area of resection. This should be the first step in preventing recurrences of these lesions, although it is dependent on tumor size, location, and the presence or absence of disease within the surgical margin. However, a literature review revealed no differences in recurrence between the patients with negative margins and those with positive margins . These rThe combination of the low metastasizing potential in AAM cases and the low mitotic activity of the tumor cells, chemotherapy and radiotherapy will have an inefficient role in adjuvant treatment . HoweverStudies have referred to a high local recurrence rate between 36% and 72%, but no metastasis has been reported to date in men . HoweverAlthough we conducted a systematic review of all cases reported in the medical literature on this topic, there are limitations to our study. For example, the available evidence remains weak because they all are case reports or series. In addition, we did not evaluate accepted studies and collected results without evaluating the evidence, but we excluded studies that did not accurately explain the method of diagnosis. However, our study remains the best available evidence about this tumor in men.6Although the occurrence of AAM is rare in men, consideration should be taken in the differential diagnosis of a mass in the genitourinary region. Radiologic images play a role in the initial approach, but an accurate diagnosis is made by the histochemistry and immunochemistry studies. According to our review, the most decisive immunohistochemistry profile is the positivity of Vimentin and CD34 with the negativity of S-100. In terms of treatment, wide surgical excision continues to be the standard. Although hormonal treatment is controversial, based on the available data and previous reports, we suggest a novel algorithm for the management of aggressive angiomyxoma .Fig. 2ThThere is no need for ethical approval.None.Aziz Joseph Sabbagh performed the surgery, followed the patient, and made a substantial contribution to the conception and design, drafting, and revision of the article.Khaled Arnaout conducted a literature search, data analysis, and made a substantial contribution to the interpretation of data, drafting, and revision of the article.Aziz Joseph Sabbagh and Khaled Arnaout contributed equally to this paper as a co-first author.Ahmad Yamen Arnaout and Bayan Toutounji made a substantial contribution to conception and design, screening the articles, extracting the data, drafting, and revision the article.Lina Ghabreau performed the immunohistochemical staining, interpreted the data, and critically revised the article.Kusay Ayoub supervised the study, interpreted the data, and critically revised the article.Ibrahim Al-Hadid supervised the study and the surgery, interpreted the data, and critically revised the article.All authors read and approved the final version of the manuscript.Written informed consent was obtained from the patient for publication of this case report and any accompanying images.OSF Registries.1. Name of the registry: 2. Unique Identifying number or registration ID: 10.17605/OSF.IO/U64WB.https://archive.org/details/osf-registrations-u64wb-v1.3. Hyperlink to your specific registration (must be publicly accessible and will be checked): Yamen.arnout@gmail.com.Ahmad Yamen Arnaout. Not commissioned, externally peer reviewed.None.


Myocardial infarction (MI) is one of the most common illnesses seriously harmful to human health. Notwithstanding, the systems of its pathogenesis are as yet not totally demonstrated. CircRNA is one sort of non-coding RNA, and late distributed information proposes that circRNAs assume a significant part in heart diseases; however, their expression profiles in the peripheral blood of patients with MI are not yet totally characterized. Therefore, RNAs from peripheral blood were recruited for high-throughput RNA-seq analysis. A total of 3,862 circRNAs were distinguished to be remarkably different, including 2,738 circRNAs being upregulated and 1,124 circRNAs being downregulated. circTMEM165, circUBAC2, circZNF609, circANKRD12, and circSLC8A1 were reconfirmed by RT-qPCR in the cell model. ROC curves uncovered that they have great sensitivity and specificity in the determination of MI. Besides, circRNAs are associated with cell metabolism and function by directing complex networks among circular RNAs, microRNAs, and messenger RNAs. In outline, our study portrayed the specific articulation profiles of circular RNA in patients with MI. The outcomes showed that circRNAs might fill in as a sort of ideal biomarkers for MI diagnosis. Further exploration of these circRNAs may enrich our understanding of MI etiology and progression. Cardiovascular diseases (CVDs) are the most common cause of death globally. The number of deaths from CVD accounted for about one-third of all global deaths . MyocardRecently, circRNAs have become the focus and the front field of RNA research based on advanced RNA sequencing technology and bioinformatics analysis. circRNA is a kind of non-coding RNA, which contains a closed continuous loop structure and is broadly expressed in multispecies genomes , 13. cirThe peripheral blood involves numerous leukocyte subpopulations, for instance, T cells, B cells, mononuclear cells, and natural killer cells. Studies have shown that key genes in leukocytes are associated with alterations in gene expression and the onset of disease .To date, the associations between circRNAs in peripheral blood and MI are not fully understood. Therefore, we explored the expression profiles of circRNA in patients with MI and healthy controls by high-throughput circRNA sequencing. We collected the volunteers’ peripheral blood and constructed the cell model to confirm the dysregulated circRNAs. ROC and combined ROC curves were created for demonstrating the biomarkers of MI. The roles of circRNAs in MI were analyzed by bioinformatics. Moreover, we discovered a possible complex endogenic RNA regulatory network among circRNAs-miRNAs-mRNAs, with a view to providing some references for related research in this field.All testing programs were performed according to the protocols approved by the Medical Ethics Committee of the Affiliated Hospital of Qingdao University. All sample providers were informed of their blood samples, clinical data were used for this study, and informed consents were signed.The peripheral blood samples from 80 patients with MI were collected before percutaneous coronary intervention surgery between March 2018 and January 2019 in the Affiliated Hospital of Qingdao University . Fresh pvia the TRIzol method . This experiment was performed according to the kit’s recommendations. RNA samples were stored at −80°C.The samples were used for RNA extraction ® Spectrophotometer . RNA concentration was detected by Qubit® RNA Assay Kit in Qubit® 2.0 Flurometer . RNA integrity was measured by the RNA Nano 6000 Assay Kit of the Bioanalyzer 2100 system . RNA degradation and contamination were monitored on 1% agarose gels.The purity of the extracted RNA was tested by a NanoPhotometer™ rRNA Removal Kit  and ethanol precipitation. Subsequently, the linear RNA was digested with RNaseR . The sequencing libraries were produced by NEBNext® Ultra™ Directional RNA Library Prep Kit for Illumina® .A total of 5 μg RNA per sample was used for the RNA library preparations. First, rRNA was removed by Epicenter RibozeroThe cluster generation of the index-coded samples was performed on a cBot Cluster Generation System using TruSeq PE Cluster Kit v3-cBot-HS. After that, the libraries were sequenced on an Illumina Hiseq 4000 platform. These experiments were performed following the manufacturer’s instructions.First, clean data were gained by wiping off reads containing adapter and low-quality reads from raw data. The clean data with high quality were used for downstream analyses. Index of the reference genome was created using Bowtie2 v2.2.8, and paired-end clean reads were united to the reference genome using Bowtie . The cirThe junction site part of circRNAs was confirmed by polymerase chain reaction (PCR) with divergent primers. Convergent primers were used as the control. The products of the PCR amplification were authenticated by 1% agarose gel electrophoresis. The primers for circRNA validation are listed in The dysregulation circRNA was reconfirmed on a CFX96 Real-Time PCR Detection System (Bio-Rad). Total RNA was extracted using Trizol reagent. Reverse transcription reactions were performed by using the TransScript II One-Step gDNA Removal and cDNA Synthesis SuperMix  to make cDNA according to the manufacturer’s guide. TransStart Green qPCR SuperMix (TransGen) was used for quantitative PCR (qPCR) analysis, and the procedures were performed in accordance with the kit instructions. The levels of dysregulation circRNA analyzed by RT-qPCR were normalized to that of GAPDH . The priThe ROC curves were created to estimate the diagnostic value of circRNAs for MI. An ROC curve was calculated, and the specificity and sensitivity of predictive power were measured by the AUC. The AUC was used to assess the diagnostic value of the circRNAs in the peripheral blood of patients with MI . The comP-value less than 0.05 were considered to be significantly enriched by differential expressed genes. As a database resource for understanding high-level functions and utilities of the biological system  enrichment analysis for host genes of differentially expressed circRNAs was carried out by the GO seq R package, and the gene length bias was corrected during this procedure . GO terml system , Kyoto Epathways .via binding to an miRNA  incubated at 37°C for 12 h in a complete culture medium. The cells were collected for RNA extraction and dysregulated circRNA reconfirmation.AC16 cells were cultured in DMEM (Gibco) with 10% fetal bovine serum (TransGen), and 100 U/ml penicillin and 100 mg/ml streptomycin (Invitrogen) in a humidified 5% COApoptosis was determined by the terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) using a kit from TransGen. The detection procedures were in accordance with the kit instructions.Mitochondrial staining was performed as described earlier with modifications . BrieflyImmunoblot was carried out as described earlier . Brieflyt-test with Welch’s correction for two groups. Statistical analyses were completed with GraphPad Prism 7.0 . P < 0.05 was regarded as statistically significant.The results are expressed as mean ± SEM of at least three independent experiments. The statistical comparison between different groups was completed by one-way ANOVA for multiple comparisons or The circRNAs were identified by find_circ and Bowtie2 based on the high-quality clean data. A total of 3,862 circRNAs were recognized in five pairs of MI and healthy controls samples. The size of the circRNAs extended from ≈100 to 2,500 nt . The difFirst, we screened candidate circRNAs by comparing our results with the previous study on human heart tissue sequencing sample results  31) in  in 31) iTo verify the RNA-seq data, five intersectional circRNAs  were selected and validated by RT-qPCR using 80 MI samples and 80 healthy individuals. The general information of selected circRNAs shown as To reconfirm the dysregulation of these circRNAs in the cell model, we used the myocardial cell injury model induced by reactive oxygen species (ROS) to simulate cell damage during MI . The datTo evaluate the potential diagnostic value of selected circRNAs for MI, we computed and created ROC curves  for RT-qThe calculated optimal cutoff point for circTMEM165 level in the peripheral blood was 1.6089505 (Youden index = 0.55), while the sensitivity and specificity were 21.3% and 23.7%, respectively. The calculated optimal cutoff point for circUBAC2 level was 1.968652 (Youden index = 0.663), while the sensitivity and specificity were 82.5% and 83.8%, respectively. The calculated optimal cutoff point for circ ZNF609 level was 1.947015 (Youden index = 0.638), while the sensitivity and specificity were 86.3% and 77.5%, respectively. The calculated optimal cutoff point for circ SLC8A1 level was 1.5414885 (Youden index = 0.737), while the sensitivity and specificity were 82.5% and 91.25%, respectively. The calculated optimal cutoff point for circANKRD12 level was 1.4872795 (Youden index = 0.575), while the sensitivity and specificity were 87.5% and 57.5%, respectively. The optimal cutoff point of the combination was 0.06, while the sensitivity, specificity, and Youden index were 0.863, 0.937, and 0.8, respectively. The results revealed that the AUC of these circRNAs at least reached 0.8073, and the AUC of recombination circRNAs reached up to 0.9289 , indicatP = 2.892E-46), intracellular part , and protein binding , respectively.The source genes of the validated differentially expressed circRNAs were uncovered by using GO annotation, as shown in The data of the KEGG pathway study of the circRNAs genes are shown in The miRNA binding sites exist in circRNAs, which can function as miRNA sponges. To further explore their possible roles in patients with MI, we predicted the circRNA-miRNA-mRNA network and analyzed using miRanda and RNAhybrid. The top five predicted miRNAs for these upregulated circRNAs are shown in Moreover, we noted the potential function of these regulatory networks. For example, circTMEM165 is predicted as “sponge” to hsa-miR-548a-3p, which can bind to mRNA GREM1, which is responsible for regulating organogenesis, body patterning, and tissue differentiation. The network of circZNF609-hsa-miR-221-5p-WDR7 may be involved in the low-density lipoprotein (LDL) metabolism. The regulatory pathway of circANKRD12-hsa-miR-876-5p-MYLK may promote myosin interaction with actin and facilitate to produce contractile activity. circANKRD12-hsa-miR-130b-5p targets both ACSM2A and RNF213, which regulate mitochondrial acyl-coenzyme A synthetase and ATPase activity, respectively. These networks indicate the possible interactional RNA regulatory relationships between circRNAs, miRNAs, and mRNAs in MI.2O2 after the knockdown of circSLC8A1 expression. We found that the number of apoptotic cells was significantly decreased in the si-circSLC8A1 group (via Western blot analysis, and our results revealed that the expression levels of Caspase3 and cyto c were markedly decreased (As circSLC8A1 has the best ROC in the verification of clinical samples, we should further explore its mechanism of action in MI. To further clarify the underlying mechanism of circSLC8A1 in MI, we first knocked down circSLC8A1 expression in the myocardial cells through circSLC8A1-siRNA. The AC16 cells were treated with 100 μM HA1 group , and theA1 group . To inveecreased . These dTaken together, these data demonstrated that knockdown of circSLC8A1 decreased intracellular oxidative stress, kept the balance of mitochondria dynamics, and then reduced apoptosis during oxidative stress.2O2. The results indicated that overexpression of circSLC8A1 remarkably promotes mitochondria fission (To understand the role of circSLC8A1 in myocardial cell apoptosis, we overexpressed circSLC8A1 in AC16 cells using circSLC8A1-plasmid. After transfection, the cell was treated with 100 μM H fission , and the fission . In cell fission . We also fission .Altogether, our results demonstrated that the overexpression of circSLC8A1 increased the intracellular oxidative stress, and it sensitized the myocardial cells to apoptosis by further increasing mitochondrial fission under the stimulation of oxidative stress.Myocardial infarction is a kind of multifactorial disease, which causes complex pathogenesis and leads to high mortality. It is hard to diagnose and prompt the intervention of MI before the disease occurs. As a consequence, there is an earnest for early detection markers for MI diagnosis. circRNAs are single-stranded, covalently closed molecules  that areRecent reports have demonstrated that circRNAs are more constant, varied, and conserved when compared with other kinds of RNAs, including mRNAs, miRNAs, and lncRNAs . circRNAEarly diagnosis can guide clinical medication to effectively treat MI and significantly improve their prognosis. circRNAs have already been confirmed as effective biomarkers for numerous diseases, such as cancer , acquireRecently, several studies have indicated that circRNAs might be key regulators in CVDs. For instance, Wu et al.  showed tWe also used circRNA-seq data to predict circRNA-miRNA-mRNA networks and aimed to find the possible roles of circTMEM165, circUBAC2, circZNF609, circANKRD12, and circSLC8A1 in the peripheral blood of patients with MI. We observed the possible target mRNAs of these circRNAs including GREM1, WDR7, MYLK, ACSM2A, and RNF213. Moreover, the predicted target mRNAs played a key role in regulating organogenesis, body patterning, tissue differentiation, LDL metabolism, and myosin interaction with actin, mitochondrial acyl-coenzyme A synthetase, and ATPase activity, respectively. All of them are highly correlated with CVDs.To further clarify the potential mechanism of circSLC8A1 in MI, we tested the cell function through knockdown and overexpressed circSLC8A1 based on the cell model. Uncontrolled mitochondrial fission contributes to apoptosis , which pA few limitations of this study ought to be mentioned. First, we only verified 10 circRNAs that were upregulated in MI, and still, a lot of differentially expressed circRNA needed to be verified. Second, although we use a set of filter methods to get the more accurate results, the sample size of this study is still limited, which will increase the false positive rate while analyzing RNASeq data. Third, we only explore more upregulated circRNAs, and the downregulated circRNAs are also worth studying in depth. Moreover, further experiments are needed to explore the regulatory effects of circRNAs in the occurrence of MI in patients with CVD.To the best of our knowledge, this study is the first to report the expression profiles of circular RNA in the peripheral blood of patients with MI compared with healthy individuals. Bioinformatics analysis indicated that the upregulated expressions of circTMEM165, circUBAC2, circZNF609, circANKRD12, and circSLC8A1 were highly correlated with MI. Moreover, all of these five circRNAs may be used to improve the diagnosis accuracy of MI. Besides, the bioinformatics analysis found that these circRNAs could be involved in the regulation of hundreds of mRNAs, and numerous of these mRNAs may affect the occurrence of MI. Although our results revealed that these circRNAs are highly expressed in the peripheral blood of patients with MI, their predicted functions still need to be confirmed by further experiments.The original contributions presented in the study are included in the article/The studies involving human participants were reviewed and approved by Ethics Committee of Affiliated Hospital of Qingdao University. The patients/participants provided their written informed consent to participate in this study.QL, JW, and YG conceived and designed the experiments. QL, YW, and YA collected and analyzed the data. QL and YW wrote this manuscript. All authors read and approved the final manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


Virtual reality (VR) has been suggested to be effective at enhancing physical exercises because of its immersive characteristics. However, few studies have quantitatively assessed the range of motion and brain activity during VR exercises.We hypothesized that 3D immersive VR could stimulate body movement and brain activity more effectively than standard exercises and that the increased range of motions during 3D immersive VR exercises would be associated with orbitofrontal activation.A randomized crossover trial was conducted to compare exercises with and without VR. A total of 24 healthy males performed the same motions when exercising with and without 3D immersive VR, and the recorded videos were used for motion analysis. Hemodynamic changes in the prefrontal cortex were assessed using functional near-infrared spectroscopy.z=−2.31; P=.02), length , calorie consumption , and change in accumulated oxygenated hemoglobin within the right orbitofrontal cortex  between the VR and offline trials. Hemodynamic changes in the right orbitofrontal cortex were positively correlated with the total angle  and length  in the VR exercise; however, there was no significant correlation in the offline trial.There were significant differences in the total angle (The results of this study suggest that 3D immersive VR exercise effectively increases the range of motion in healthy individuals in relation to orbitofrontal activation.Clinical Research Information Service KCT0008021; https://cris.nih.go.kr/cris/search/detailSearch.do/23671 Virtual reality (VR) is an artificially created sensory experience using digital technology, which provides users with a subjective sense of “being there” . The forVR is expected to have enormous potential in fields such as education, training, and entertainment . In partVR application in sports can increase motivation, enjoyment, and physical performance . This maThe shortcomings of physical exercise using an immersive VR have been attributed to the use of HMDs. Exercising while wearing an HMD can be uncomfortable due to sweating and head bumps . In addiVR exercises are effectively used in the rehabilitation of patients who have lost motor skills ,14. TheyPrevious studies have suggested that the brain regions of interest in response to VR and physical exercises are the dorsolateral prefrontal cortex (DLPFC), ventrolateral prefrontal cortex (VLPFC), and orbitofrontal cortex (OFC) ,21,22. OThis study aimed to investigate whether immersive VR technology can enhance physical exercise, in terms of the range of motion and brain activity, in healthy individuals. Kinematics and resting state fNIRS methodologies were adopted to demonstrate the quantitative effectiveness of 3D immersive VR exercise. Kinematics is useful for objectifying human movements and comparing different subjects or situations, such as before and after treatment or training in various fields  . For exaWe hypothesized that 3D immersive VR exercises would be more effective than offline physical exercises in increasing the range of motion and calorie consumption during exercise. In addition, we hypothesized that the effectiveness of 3D immersive VR exercises is associated with brain activation within the OFC before and after exercise.The sample size was calculated using G*power software (Heinrich-Heine-Universität Düsseldorf)  based onThrough flyer advertisements, 24 healthy males were recruited for this study from Chung-Ang University. The inclusion criteria were as follows: (1) male sex, (2) age 20 to 29 years, and (3) no psychiatric or medical illness. The exclusion criteria were as follows: (1) history of head trauma; (2) history of substance abuse, including alcohol, tobacco, and drugs; and (3) IQ<80. The participants were instructed not to consume any food, caffeine, or alcohol 3 hours prior to participation in the study.The institutional review board of Chung-Ang University approved this research protocol (1041078-201908-HRSB-231-01). Written informed consent was obtained from all participants.This study was a randomized crossover trial that compared responses to 3D immersive VR and offline exercises. The participants were blinded to the study hypotheses. Each participant underwent 1 3D immersive VR trial and 1 offline trial in a randomly assigned order at 10-minute intervals. Each session consisted of (1) a 2-minute practice to become familiarized with each exercise; (2) a 2-minute pre-exercise resting-state fNIRS measurement; (3) a 5-minute exercise, either 3D immersive VR or offline; and (4) a 3-minute postexercise resting-state fNIRS measurement . This trBoth VR and offline exercises consisted of 40 identical movements derived from Pilates. Pilates was chosen because it is a beneficial exercise method for healthy adults that enhances flexibility, muscular activity, and coordination . In bothBefore the VR exercise, the research staff provided brief instructions on the 3D immersive VR system and environment and asked the participants to report whether they experienced any discomfort while wearing the HMD, such as head bumps, sweating, or motion sickness. The research staff checked for any obstacles the participants might encounter during the VR exercise and monitored and provided immediate assistance to the participants during the exercise in case of danger. With the help of the research staff, the participants were positioned 2 m horizontally away from the VR device, wearing the HMD and holding resistance bands in both hands at shoulder level.In this study, the Vive Cosmos Elite (High Tech Computer Corp) and Kinect V1 (Microsoft) were used as the HMD and motion sensors, respectively. A 3D map–based immersive VR software system that allows participants to interact with the VR environment was developed using Unity3D Engine (Unity Technologies). The Grand Canyon landscape was designed to enable participants to fly between canyons without crashing by performing the aforementioned movements in sequence. During the VR exercise, participants were asked to fly over the canyon without bumping into it, following visual and audio guidance. The direction in which to move, whether to accelerate or decelerate, and flight speed were displayed on the VR screen in real time, serving as visual instructions . When thIn contrast, during the offline exercise, the participants performed movements in a standard manner following a demonstration by a professional instructor. In both sessions, audio guidance, such as “lean right and raise your left arm” were provided.The research staff helped the participants accurately perform each movement in both sessions. They provided immediate feedback for incorrect movements and high-risk situations. The movements performed by the participants in the VR and offline conditions were considered identical since the audio guidance, use of the resistance band, and real-time feedback on behavior by the research staff were the same for all participants.Participants’ movements during the VR and offline exercises were recorded using an iPhone 11 camera (Apple). The participants were asked to stand at the designated site 1.5 m away from the camera to maintain a constant camera angle. The recorded video was analyzed using the open-source video analysis software Kinovea  . KinoveaIn the motion analysis, eight types of motion angles were measured: (1) right waist angle, (2) left waist angle, (3) right body angle, (4) left body angle, (5) right leg angle, (6) left leg angle, (7) forward body angle, and (8) backward body angle. Additionally, four types of movement lengths were measured: (1) right arm movement length, (2) left arm movement length, (3) right leg movement length, and (4) left leg movement length . The totCalorie consumption during each session was calculated using each participant’s weights, motion retention time (from the start to the end of the movement), and the metabolic equivalent of task (MET) of the Pilates motion, according to the MET formula . The METHemodynamic changes within the prefrontal cortex were assessed using a high-density fNIRS device . The curved panel of the NIRSIT has 24 laser diodes (sources) emitting 2 wavelengths (780 nm and 850 nm) of light and 32 photodetectors with a sampling rate of 8.138 Hz. The unit distance between the source and photodetectors was 15 mm. In this study, only the 30-mm channels were analyzed, since 30 mm is the most suitable sensor-detector separation distance for measuring cortical hemodynamic changes.The fNIRS data were processed using the analysis toolbox provided by the fNIRS device manufacturer. The light signals in each wavelength within the 48 channels were filtered using a band-pass filter (between 0.0 Hz and 0.1 Hz) to reduce noise due to external light and body movements. Data derived from channels representing low-quality information  were not included in the hemodynamic analysis set to prevent misinterpretation. Relative hemodynamic changes during exercise were calculated using the modified Beer-Lambert law . The accThe values (mean and SD) of accHbO2 were gathered from 8 regions of interest: the right and left DLPFC, right and left frontopolar cortex (FPC), right and left VLPFC, and right and left OFC. The right and left DLPFCs were composed of channels 1, 2, 3, 5, 6, 11, 17, and 18 and channels 19, 20, 33, 34, 35, 38, 39, and 43, respectively. The right and left FPC were composed of channels 7, 8, 12, 13, 21, 22, 25, and 26 and channels 23, 24, 27, 28, 36, 37, 41, and 42, respectively. The right and left VLPFCs were composed of channels 4, 9, and 10 and channels 40, 44, and 45, respectively. The right and left OFCs were composed of channels 14, 15, 16, 29, and 30, and channels 31, 32, 46, 47, and 48, respectively .U test. The statistical significance of the total angle and total movement length in the total behaviors was set at 0.02 . In the post hoc test, the statistical significance of the total angle and total movement length in the simple behaviors, 2 complex behaviors, and 3 complex behaviors was set at P=.02 . Between the VR and offline situations, the difference in the change in accumulated oxygenated hemoglobin (ΔaccHbO2) from baseline and exercise was compared using repeated measures ANOVA. The statistical significance was set at P=.006 . Correlations between the differences in accHbO2 within the right OFC, total angles, and total movement lengths were assessed using Pearson correlation coefficient. Statistical significance was set at P=.03 . Statistical analyses were performed using SPSS software .For total behaviors, simple behavior, 2 complex behaviors, and 3 complex behaviors, the differences in total angles, total movement length, and consumed calories were compared using the Mann-Whitney z=−2.31; P=.02) and length between the VR and offline trials . The VR group showed a 13.3% wider total angle range and an increase of 14.1% in the total movement length compared with the offline group. In the post hoc analysis, the VR group moved significantly more in simple and complex behaviors, except for the total angle in 2 complex behaviors, compared to the offline group . During exercise, the VR group consumed 61.7% more calories than the offline group. In the post hoc analysis, the VR group showed significant calorie consumption with 2 complex and 3 complex behaviors compared to the offline group  but not within the left VLPFC , left DLPFC , right DLPFC , right VLPFC , left OFC , left frontopolar prefrontal cortex , or right frontopolar prefrontal cortex   .r=0.36; P=.005). However, it was not significantly associated with the total movement length . In the VR group, the ΔaccHbO2 within the right OFC was positively correlated with total angle  and total movement length . In the offline group, the ΔaccHbO2 was not significantly correlated with total angle  or total movement length   .Of the 24 participants, 3 (12.5%) reported motion sickness, and 8 (33.3%) reported sweating after wearing the HMD device; however, none reported HMD slipping or severe discomfort.In this study, VR exercise increased the range of motion, calorie consumption, and brain activity within the OFC compared with offline exercise. In addition, increased brain activity within the right OFC was correlated with an increased range of motion.Our findings are consistent with previous studies on various patient groups. However, no studies have reported changes in the range of motion due to VR exercise in healthy individuals, as in this study . VR exerThe larger range of motion and caloric consumption in the VR exercise may be associated with immersion. Immersion can be divided into 3 forms: sensory, imaginative, and challenge-based . SensoryAnother important finding of our study was that there were significant differences in brain activation within the right OFC between offline and exercise groups. To date, only a few studies have specifically investigated orbitofrontal activity in response to VR exercises. Most studies have focused on prefrontal activation caused by VR stimuli. Mao et al  suggesteThe results of the correlation study between ΔaccHbO2 within the right OFC and the range of motion during VR and offline exercise showed that an increase in the range of motion and right OFC activation was correlated only in the VR group but not in the offline group. Thus, we suggest that immersion and visual stimulation of 3D immersive VR distracts the user from unpleasant interoceptive sensations caused by physical exercise. It is suggested that distraction caused by VR can reduce the perception of physical discomfort through an intercortical modulation of the anterior cingulate and OFC ,58,59.Additionally, the functional hemispheric differences shown in the results of this study are in line with a previous study that suggested that the more pronounced the shift in brain activity to the right hemisphere, the more flow experienced by elite tennis players . Less syTo the best of our knowledge, this is one of the earliest studies to quantitatively evaluate the effects of 3D immersive VR exercise on brain hemodynamics in healthy individuals. Each participant’s movement was measured using professional video analysis software, unlike other studies based on subjective performance improvement or satisfaction. Another strength of this study is that we directly compared performance in 3D immersive VR and real-world environments while performing the same activities. This comparison is important to reveal how 3D immersive VR influences exercise performance and brain activity. Finally, this study presents the potential for developing smart headsets by integrating wearable devices and connected technologies, such as VR, brain activity measurement, and motion tracking. Although these were performed with different devices in our study, using an integrated smart headset could enable effective exercise and immediate checking of the exercise amount and the consequent changes in brain activity. These wearable technologies and connected solutions with VR capabilities are already widely adopted in the workplace to promote occupational safety, productivity, and worker health. Examples include the Xsens MVN Animate and Xsens MTw Awinda for body movement tracking and the Muse 2 Melon's headband and IMEC's wireless electroencephalogram headset for brain wave sensing .This study had some limitations. First, due to the small number and gender disparity of the participants, the results cannot be generalized. In a follow-up study on VR exercise, scaling up the sample size by including virtual participants could be considered. However, this was not possible in this study due to the crossover design comparing VR and offline exercise. Second, there were limitations in the adequate pixel rate, capture rate, maximum aperture, depth of field capture, and frame capture rate of the iPhone 11, which was used as a video recording device. Third, since the fNIRS device used in this study can only measure brain activity within the prefrontal lobe, activity in other cortical regions and deep brain areas was not measured. Further studies using other functional imaging devices, such as fMRI, are required to determine the activity of the entire brain. Fourth, we did not employ additional physiological measures. The use of systemic physiologically augmented fNIRS in future studies may yield more meaningful results. Finally, a standardized questionnaire to check the inclusion and exclusion criteria and to assess motion sickness after VR exercise was not used in this study.This study confirmed that 3D immersive VR exercise is more effective at increasing the range of motion and calorie consumption during exercise in healthy individuals via activation within the OFC.


CDC Museum COVID-19 Timeline to pair collected data from our studies with specific moments in the pandemic—from its known origins to springtime 2022. Results highlighted how slow and deleterious forms of racist violence could wear and tear at the wellbeing of targeted people of color. Overall, this research underscores the possible hidden harms associated with slow-moving forms of racism, as well as some of the unseen stressors experienced by people of color living in the United States.Racism against people of Asian descent increased by over 300% after the COVID-19 pandemic outbreak in the United States, with one in five Asian Americans reporting direct experiences with overt discrimination. Large-scale efforts and resources initially, and quite understandably, prioritized investigating the physiological impact of the coronavirus, which has partially delayed research studies targeting the psychological effects of the pandemic. Currently, two studies tracked the unique relationships between psychosocial factors, such as experiencing everyday racism, and the self-reported wellbeing of Asian Americans in the United States and compared these associations with Latinx Americans. Study 1 (April 2020–April 2021) examined how Asian and Latinx Americans varied in their levels of wellbeing, fear of the coronavirus, internalized racism, and everyday experiences with racism. Study 2 (September 2021–April 2022) included the same variables with additional assessments for victimization distress. We used the  Asian American to include all people of Asian descent residing in the United States. In addition to this, as suggested by the American Psychological Association (APA)'s best practices for bias-free language  explored the effects of internalized racism, everyday racism, and fear of the coronavirus on overall wellbeing during the time frame of the nationwide lockdown and the first vaccine release that occurred. Study 2 (N = 185) examined the dynamics between the same psychosocial factors with additional assessments of victimization distress during the months following the widespread availability of COVID-19 vaccines in the United States.Thus, the current two studies tracked the relationships between different psychosocial factors and the wellbeing of Asian and Latinx Americans. Due to the dynamic nature of the pandemic, cross-sectional designs were used to evaluate multiple psychological factors related to racism, fear of the coronavirus, and their associations with one's wellbeing at different time points of the pandemic. This approach allowed for timely data collection without the additional resources needed for a longitudinal design; at the start of this project, it was also unclear how long the pandemic would last. Moreover, we incorporated novel measurements developed specifically for the pandemic, such as COVID-related victimization distress. Study 1 (CDC Museum COVID-19 Timeline (Study 1 (20 April 2020–27 April 2021) examined the relationships between psychosocial factors of everyday racism, internalized racism, and fear of the coronavirus with overall wellbeing among Asian and Latinx Americans. According to the Timeline , the firTimeline  testifieGiven this dramatic shift in current anti-Asian sentiments, it was hypothesized that Asian Americans would be more negatively impacted overall during the pandemic with lower overall wellbeing, more exposure to everyday racism, more internalized racism, and more fear of the coronavirus than Latinx Americans. Moreover, it was hypothesized that overall wellbeing would be strongly associated with everyday racism, internalized racism, and fear of the coronavirus for Asian Americans, compared with Latinx Americans, that is, race would moderate the relationships between these psychosocial factors and wellbeing.Mage = 19.68, SD = 2.83) and a gender breakdown of 68% women, 31.1% men, and 0.9% “other.” The overall racial breakdown of the sample was 48.1% Asian American and 51.9% Latinx American. The sample included 71 foreign-born and 284 U.S.-born participants. There were 11 participants identified as international students. A total of six participants were removed for residing outside the United States. See Data for study 1 were collected online during the COVID-19 pandemic, with all participant responses coming from a diverse respondents from Southwest University in the United States. Participants were recruited from an undergraduate psychological participant pool and were compensated course credit for their participation in the study. The final sample in this study consisted of 366 respondents between 18 and 47 years , along with participants' age, gender, socioeconomic status (SES), and generational status. These demographic questions were delivered at the end of the study to avoid possible priming effects. As previously mentioned, the primary objective of this study was to examine whether there were significant racial differences across different psychosocial factors, particularly related to one's race and the pandemic, for Asian and Latinx Americans during the early stages of the COVID-19 pandemic. Only monoracial people were included in this study since the main study variables were race-related.at no time) to 5 , with a possible raw score ranging from 0 to 25. The general scoring rule for the WHO-5 is multiplying the raw score by 4 so that the final score range is between 0  and 100 . Topp et al.   consistsp et al.  conducte samples  and has  samples , 41. Wannever) to 4 (often). Another major focus of this study was to explore the perceived shift in racism-related experiences during the COVID-19 pandemic. Therefore, we modified each EDS item with the words “before the COVID-19 pandemic” and “during the COVID-19 pandemic” to assess perceived experiences with everyday racism during the time frame. Higher scores indicated more consistent experiences with everyday discrimination. Previous studies have shown good internal reliability of the EDS, with α = 0.93 for Latinx American samples   consists samples  and α =  samples . In studstrongly disagree) to 7 (strongly agree). Higher scores suggest greater levels of internalized racism. The ER-AROS showed high internal reliability with samples of Latinx and Asian Americans, with α = 0.83   is a 7-iα = 0.83 . Similarα = 0.83 . In studstrongly disagree) to 5 (strongly agree). The scores were summed to reflect an overall score ranging from 7 to 35. Higher scores indicated a greater fear of the coronavirus. The FCV-19S showed high internal reliability for Latinx Americans, with α = 0.87   consistsα = 0.87 , and Asiα = 0.87 . Study 1Partial correlation analysis along with analysis of variance (ANOVA) of the demographic characteristics and study variables was used to examine differences among the sample while holding everyday experiences with racism before the pandemic constant see . CompariAnalysis of covariance (ANCOVA) was conducted on the main outcome and moderating variables—wellbeing, everyday racism, internalized racism, and fear of COVID-19—while controlling for a participants' gender, generational status, and SES. Perceived experiences of everyday racism before the COVID-19 pandemic were also controlled for to better assess the shift in participant experiences during the pandemic.SPSS PROCESS Macro  was usedF = 27.25, p < 0.001, F = 6.84, p = 0.02, p = 0.07. ANCOVA suggested that Asian and Latinx Americans were not significantly different in their levels of wellbeing when controlling for everyday racism before the pandemic and the other covariates. Asian Americans, however, did report higher levels of internalized racism, R2 = 0.10, F = 5.96, p < 0.001. See The three moderating analyses assessed the role of race on the relationship between psychosocial factors  and an individual's overall wellbeing. The results showed that race (Asian vs. Latinx Americans) was a significant moderator for the effects of everyday racism and fear of the coronavirus but not internalized racism. Notably, more everyday racism and heightened fear levels of the coronavirus negatively impacted the wellbeing of Asian Americans, but not Latinx Americans. For the relationship between everyday racism and wellbeing, the main independent variable (everyday racism) and the moderator (race) accounted for a significant amount of variance in wellbeing, R2 = 0.01, ΔF = 4.26, p = 0.04, on the relationship between everyday racism and wellbeing, b = −0.80, t = −2.06, p = 0.04, 95% CI . Furthermore, an examination of the simple slopes showed a crossover interaction with a negative relation for Asian Americans and a positive relation for Latinx Americans, that is, more experiences of everyday racism for Asian Americans showed lower wellbeing, whereas more experiences of everyday racism for Latinx Americans showed higher wellbeing. To avoid potential multicollinearity with the interaction term, the variables were centered, and an interaction term between everyday racism and race was created . The resR2 = 0.12, F = 7.21, p < 0.001. See For the relationship between fear of the coronavirus and wellbeing, the main independent variable (fear of coronavirus) and the moderator (race) accounted for a significant amount of variance in wellbeing R2 = 0.02, ΔF = 7.33, p = 0.01, on the relationship between fear of the coronavirus and wellbeing, b = −0.80, t = −2.06, p = 0.04, 95% CI . An examination of the slopes showed a significant effect for Latinx Americans, b = −0.84, t = −3.47, p = 0.001, 95% CI , but not for Asian Americans, that is, more severe fear of the coronavirus for Latinx Americans was associated with significantly lower overall wellbeing. The results revealed a significant moderating effect of race, ΔStudy 2 (15 September 2021–29 March 2022) compared Asian and Latinx Americans during the months after the vaccine release and the lifting of major nationwide lockdown mandates. This study included the same measures as study 1, as well as additional assessments for distress over coronavirus victimization. Study 2 began data collection during a time when CDC studies provided further evidence that COVID-19 vaccines offered higher protection than a previous COVID-19 infection alone.Mage = 19.70, SD = 3.96), with a gender breakdown of 68.0% women, 31.1% men, and 0.9% “other.” The overall racial and ethnic breakdown of the sample was 47.0% Asian Americans and 53.0% Latinx Americans, which was similar to the distribution seen in study 1. Asian American participants reported significantly higher socioeconomic status than Latinx American participants. The sample included 41 foreign-born and 144 U.S.-born participants, and five participants were international students. All participants in the final sample resided in the United States during the pandemic and understood written English. A total of five participants were removed from the initial 158 participants for either being younger than 18 years or residing outside the United States. Data for study 2 were collected online during the COVID-19 pandemic from the same university as in study 1. The final sample in this study consisted of 185 respondents between 18 and 50 years .Refer to study 1 Measures section for information on everyday racism, internalized racism, and fear of the coronavirus scales.Specific racial and ethnic backgrounds were assessed as part of the demographic questionnaire, along with age, gender, SES, and generational status. Similar to study 1, these demographic questions were delivered at the end of the online questionnaire to avoid any possible priming effects. In addition, only monoracial people were included in this study since the main study variables were race-related.it never happened) to 5 (it happened and upset me quite a bit). Fisher et al.   was usedr et al.  validatePartial correlation analysis along with analysis of variance (ANOVA) of test the demographic characteristics and study variables was used to examine differences among the sample while holding everyday experiences with racism before the pandemic constant see . SimilarANCOVAs were conducted on the same main outcome and moderating variables as in study 1: wellbeing, everyday racism, internalized racism, and fear of the coronavirus. Furthermore, we examined distress related to COVID-19 victimization, using the same covariates as in study 1. We used SPSS PROCESS Macro  to test The comparison analyses show that Asian and Latinx Americans varied across gender, generational status, and SES, but not age see . ConsideF = 6.31, p = 0.01, F = 4.14, p = 0.04, Similar to study 1, comparison ANCOVAs indicated Asian and Latinx Americans did not differ significantly on wellbeing. Unlike study 1, there were no racial differences with internalized racism and everyday racism, when controlling for the mentioned covariates. Nevertheless, Asian participants did report more fear of the coronavirus, F = 6.93, p = 0.01, R2 = 0.16, F = 3.86, p < 0.001. See Moderating analyses suggested that race did not significantly moderate the relationships among everyday racism, internalized racism, fear of coronavirus, and wellbeing. Despite that result, race moderated the effect of distress from coronavirus victimization on wellbeing, R2 = 0.03, ΔF = 5.35, p = 0.02, on the relationship between coronavirus victimization distress and wellbeing, b = 1.96, t = 2.31, p = 0.02, 95% CI . A closer examination of the simple slopes showed that higher levels of distress negatively correlated with wellbeing among Latinx Americans, t = −3.00, p = 0.003, 95% CI , but Asian Americans did not share those effects. This finding is slightly contradictory to our predictions, given that the relationship was stronger and more significant for Latinx Americans than for Asian Americans. To avoid potential multicollinearity with the interaction term, all primary variables were centered, and an interaction term between the distress of coronavirus victimization and race was created . The resUndoubtedly, the COVID-19 pandemic—from the beginning to the end—was a psychologically distressing time for many people. To this point, findings from both studies demonstrate that Asian and Latinx Americans experienced reduced levels of wellbeing during the recent pandemic; however, no significant racial differences in wellbeing were observed between the groups. Nevertheless, it is worth noting that nearly half of the Asian and Latinx Americans from the samples reported reduced levels of wellbeing in general, with almost one in six meeting the cutoff for clinical depression in study 1 and almost one in 10 in study 2. Despite the similarities between Asian and Latinx Americans, their reduced levels of wellbeing, which suggested probable depression in many cases, were nearly two times higher than the rates seen in the general population before the pandemic.This sudden drop in wellbeing and rise in depressive rates warrant further attention from U.S. officials since it seems to indicate a growing public health concern for Asian Americans, as well as Latinx Americans. Another point worth noting is that for Asian Americans, rates of reduced wellbeing were climbing between the time frames of the studies (43.8–49%), whereas for Latinx Americans, rates of reduced wellbeing were lowering (52.6–40.8%). This considerable reduction in wellbeing, especially if left unaddressed, might increase Asian Americans' susceptibility to psychological disorders and problematic behaviors, such as depression and addictions. In turn, Asian Americans might seek out healthcare services at a greater frequency and at much higher rates than before the pandemic. Barriers to cultural competent services  may need additional improvements in regard to assisting Asian Americans in dealing with the aftermath of the pandemic.Furthermore, the fast and attention-grabbing violence toward Asian Americans that was widely publicized during the pandemic, corroborating the findings of study 1 that indicated a significant upward shift in everyday racism for Asian Americans as compared with Latinx Americans. Asian Americans also reported relatively higher levels of internalized racism throughout the duration of the studies. The exponential uptick of hate crimes, or fast violence, against Asian Americans during the pandemic perhaps spilled over into the racism-related slow violence reported by the participants in both the studies, based on their everyday exchanges, internalized hate, and/or victimization distress. In addition, moderating analyses showed that an individual's race was a significant moderator for the relationship between everyday racism and overall wellbeing. Not only was there an upward shift in experiences of everyday racism but also this effect seemed more negatively linked to Asian Americans' levels of wellbeing. This general trend in both the studies highlights the unique burden that racism places on many people of color. Latinx Americans may have not experienced more distress from COVID-19-related victimization, but Latinx Americans did report more distress and lower wellbeing than Asian Americans.In response the racial inequities during the pandemic, the APA issued a call for an increase in training, awareness, research, and the creation of clinical tools that are culturally adapted for people of color , 50. OneIn comparison to Latinx Americans, Asian Americans reported more worry about the coronavirus throughout the pandemic, particularly in relation to the fear of infection. This increase in generalized fear and anxiety, along with navigating the blindsiding shift in racism, plausibly made many Asian Americans more susceptible to coronavirus infections and other ailments. In 2020, one in seven Asian American deaths were related to COVID-19, and Asian Americans were overrepresented in mortality rates compared with non-Hispanic whites . EmerginDifferences in racism-related factors were not significant when the COVID-19 vaccines were widely available during study 2. As predicted, Asian Americans reported more distress from coronavirus victimization than Latinx Americans. Much of the distress Asian Americans experienced appeared to be more related to being blamed for the coronavirus outbreak. Overall, though, these results suggest that the psychological impact of COVID-19 might be, by comparison, more robust for Asian Americans; however, Latinx Americans who experience more fear and blame for the coronavirus experienced lower levels of wellbeing. During the later months of the pandemic, major efforts were launched to stop hate against Asian American communities in the United States . Perhaps these public gestures to stop racism against Asian Americans in the United States also mitigated some of the everyday and internalized racism effects, whereas Latinx Americans did not experience the same large-scale efforts during the pandemic.We examined the factors related to psychosocial experiences with racism among Asian Americans and the coronavirus during the critical months of the pandemic as compared with Latinx Americans. The results from these studies should be interpreted considering research limitations. First, the data were collected as online questionnaires from two college samples at the same university. Therefore, generalizability interpretations of our findings might be limited to an emerging adulthood experience, particularly young people completing an undergraduate degree. Future studies should assess other demographic groups since many effects of pandemic may continue to linger throughout the United States. In addition, our studies were cross-sectional designs examining separate samples at different time points of the pandemic. These studies only considered associations between psychosocial factors and wellbeing, so future longitudinal methodologies will help clarify the directionality of these effects.can be weapons—ones that often inflict emotional harm and pain far beyond what the human eye is able to observe. In our samples, we were able to glimpse some of these negative effects that racist and xenophobic words and acts can have on Asian American people, many of whom experienced more consistent fear and distress during the time of the pandemic. Lately, it seems most instances of fast violence against Asian Americans might have, for a time, been mitigated, but the unseen effects of racism will likely endure for many people of color.In sum, the takeaway message from the current findings is that Asian Americans have experienced and still are experiencing multi-faceted forms of racism. During the beginning months of the pandemic, Asian Americans in our samples reported significantly more experiences with the slow violence of racism than Latinx Americans; and while some people may shutter at the very thought of calling words a form of violence, this term is meant to capture an important truth: hateful words The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.The studies involving human participants were reviewed and approved by Institutional Review Board. The patients/participants provided their written informed consent to participate in this study.GW-P was lead author and contributed to the conception, design, organized the database, performed statistical analyses, and wrote the draft of the manuscript. AB contributed to the conception and design of the study, and organized the database. AK and MS wrote sections of the manuscript. All authors contributed to the manuscript revision, read, and approved the submitted version.The publication fees for this article were supported by the UNLV University Libraries Open Article Fund.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


Keratoconjunctivitis sicca (KCS) is predominantly an immune-mediated chronic inflammatory ocular disease that is commonly diagnosed in dogs. This study aimed to compare the conventional use of topical immunosuppressant tacrolimus 0.03% eye drops and a new therapy injectable homologous platelet-rich plasma (HPRP) into the third eyelid gland and inferior and superior palpebral conjunctiva of dogs with KCS.A total of 66 eyes from 33 dogs were evaluated. The eyes were divided into three equal groups: Negative control group, tacrolimus group (TG), and homologous platelet-rich plasma group (HPRPG). The animals were evaluated using the Schirmer’s tear test-1 (STT-1), osmolarity test (OT), strip meniscometry test (SMT), tear film break-up test (TBUT), fluorescein test, lissamine green test (LGT), and cytological and histopathological analyses.In TG, there was a significant increase (p < 0.05) in the STT-1 and SMT values, and goblet cell count in the palpebral conjunctiva by the end of the study. In HPRPG, 36% (four dogs) received three applications, 55% (six dogs) received two applications, and 9% (one dog) received one application before the initial ocular signs improved. There was a significant decrease (p < 0.05) in the lymphocyte and neutrophil counts of the palpebral conjunctiva in HPRPG than in TG. Both groups showed equivalent improvements in TBUT, OT, and LGT values.Tacrolimus 0.03% eye drops were more efficient than HPRP in increasing tear production and the number of goblet cells. However, injectable HPRP was more efficient than tacrolimus in decreasing the number of conjunctival inflammatory cells. Treatment with injectable HPRP requires an average of two to three applications, is safe and feasible, and can be used as a cheaper alternative or as an adjuvant to conventional treatment with topical immunosuppressants. Keratoconjunctivitis sicca (KCS), also known as dry eye disease, is characterized by chronic inflammation of the tear glands, causes decreased tear production (quantitative KCS) or deficiency in tear quality , or both. Immune-mediated KCS is the most common type in dogs and humans –3. The mThe following tests are used in the diagnosis of KCS: Schirmer’s tear test (STT-1), tear film break-up test (TBUT), tear osmolarity test (TOT), Lissamine Green test (LGT), Rose Bengal test, phenol red test, evaluation of the meibomian glands and tear film, cytological exam, and histopathological exam of the conjunctiva . SchirmeStreptomyces tsukubaensis spp. that causes the proliferation of goblet cells. It inhibits the immune response by binding to FK506-binding protein, which impedes the initial phase of type-T lymphocytes activation. The intraocular action of tacrolimus can be variable and potentiated according to the concentration and transport vehicle used [Conventional treatment includes a combination of topical drugs, artificial tears or ocular lubricants, and calcineurin inhibitors that have tear-stimulating and immunosuppressive effects, such as tacrolimus, cyclosporin A, and pimecrolimus. These drugs suppress interleukin-2 production, prevent the formation of inflammatory cascade and eosinophil recruitment, and reduce inflammation by inhibiting histamine release and prostaglandin production –15. Tacrcle used –17.Platelet-rich plasma (PRP), a byproduct derived from blood plasma, has tissue-reconstituting and reparative properties since it contains growth factors, cytokines, integrins, and Vitamin A. Platelet-rich plasma is rich in bioactive proteins synthesized in the blood, mainly platelet alpha granules. It has an immunomodulatory function, can be easily obtained and used, and is inexpensive . In patiPlatelet-rich plasma , homologous (extracted from the same species), or heterologous (extracted from another species) in origin. Homologous and heterologous forms of PRP are only used when autologous PRP is unavailable. First, the whole blood of the donor is centrifuged, with the addition of anticoagulants such as thrombin or sodium citrate. Sedimentation occurs during this process, leaving the platelets suspended in the plasma at a concentrated level; the growth factors are then released. The process of obtaining the PRP takes an average of 1 h. The platelet concentration in PRP is usually 4–7 times higher than that in whole blood –21.This study aimed to evaluate and compare two treatment methods for KCS: A new treatment by injecting homologous PRP (HPRP) into the third eyelid gland, and lower and upper conjunctiva versus conventional treatment with tacrolimus 0.03% eye drops, a topical immunosuppressant.The study was approved by the Ethical Committee on Animal Use of Universidade do Oeste Paulista (UNOESTE - Protocol No. 5726) and was conducted according to the Association for Research in Vision and Ophthalmology guidelines for the use of animals in ophthalmic and visual research.The study was conducted from August 2019 to July 2021 in Veterinary Hospital (VH) and kennel at UNOESTE, Presidente Prudente, São Paulo, Brazil.Dogs diagnosed with KCS were identified during routine clinical visits to the ophthalmology service of the VH of the UNOESTE. The dogs in the negative control group (NCG) were selected from the kennel at UNOESTE. A total of 33 dogs (66 eyes) of both sexes and different breeds, aged 7.5 ± 3.6 (1.0–15.0) years and weighing 11.7 ± 10.5 (3.6–43) kg, were included in the study. The animals were divided into three groups (NCG and two KCS treatment groups). The inclusion criteria for the study were as follows: For healthy eyes – rate of lacrimation on STT-1 without anesthetic ≥15 mm/min, TBUT of >20 s, and no remarkable findings on ophthalmic examinations; and for eyes with KCS – presence of ophthalmic clinical signs , lacrimation rate on STT-1 <15 mm/min or TBUT <20 s, or both. The guardians of each animal signed an informed consent form. All ocular tests and examinations were performed by the same examiner (GJGE).Three groups of dogs, each with 11 animals (22 eyes), were evaluated: the NCG contained dogs who did not have KCS; dogs that were diagnosed with KCS were randomly assigned to either the tacrolimus group (TG) or homologous PRP group (HPRPG). The characteristics of the groups are described below.Negative control group : 11 healthy dogs ; aged 4.3 ± 1.9 (1.0–7.0) years; weighing 20.4 ± 14.9 (5.0–43.0) kg; nine mixed-breed dogs, one Boxer, and one Labrador. The NCG was used to extract reference ocular values for the study region and to serve as a comparison group with the KCS-positive groups.Tacrolimus group (n = 22 eyes positive for KCS): 11 dogs ; aged 8.5 ± 2.1 (5.0–11.0) years; weighing 7.5 ± 1.4 (5.2–9.8) kg; seven Lhasa Apso, one Shih Tzu, one Yorkshire Terrier, and one mixed-breed dog. Once the diagnosis of bilateral KCS was confirmed, the following protocol was established: topical treatment with 0.03% tacrolimus eye drops , at one drop twice a day in both eyes, and eye lubricant based on 20% chondroitin A sulfate , at one drop twice a day, in both eyes, for 6 months.Homologous platelet-rich plasma group (n = 22 eyes positive for KCS): 11 dogs ; aged 9.7 ± 4.0 (5.0–15.0) years; weighing 7.1 ± 2.4 (3.6–12.1) kg; six Lhasa Apso, two Shih Tzu, two Yorkshire Terriers, one Cocker Spaniel, and one mixed-breed dog. Once the diagnosis of bilateral KCS was confirmed, the following protocol was established: In the ambulatory clinic, one drop of anesthetic eye drops  was instilled in each eye topically, 3 times at an interval of 5 min; then, 0.3 mL of HPRP was administered with an insulin syringe and needle at 0.1 mL in the third eyelid gland, 0.1 mL in the inferior palpebral conjunctiva, and 0.1 mL in the superior palpebral conjunctiva of both eyes . In addiIf diagnosed with secondary bacterial infection and ocular inflammation (conjunctivitis or keratitis) in TG and HPRPG, ciprofloxacin-based antibiotic eye drops (Laboratory Labyes), at one drop 3–4 times a day and sodium diclofenac-based anti-inflammatory eye drops (Allergan), at one drop 3 times a day for 15 days, were prescribed , 22, 23.Homologous platelet-rich plasma was prepared at the clinical pathology laboratory of the VH of UNOESTE. Only one healthy 5-year-old (weight 35 kg) of mixed-breed was used as the donor dog. In detail, 20 mL of whole blood was collected in sterile tubes containing 3.2% sodium citrate using the vacuum system for each animal that received therapy by puncturing the jugular vein. In the first stage, the blood was centrifuged at 200 RCF or 1,100 RPM for 10 min in a laboratory centrifuge . Approximately 30% of the surface plasma was discarded, and the remaining content was transferred to a dry and sterile Falcon tube, which was then centrifuged at 400 RCF or 1900 RPM for 10 min. At this stage, the plasma was divided into two portions, PRP and platelet-poor plasma. Two-thirds of the supernatant material was discarded (platelet-poor plasma), and the remaining one third was classified as PRP. A total volume of 0.6 mL was obtained from this material, and 0.1 mL was instantly applied to the third eyelid gland, and lower and upper eyelid conjunctiva .Ophthalmic signs were evaluated monthly during 6 months of treatment, from moment M0 (time of diagnosis of KCS) to M6 . The ophthalmic signs of each animal were evaluated using a portable slit lamp ; a specific form was used to evaluate corneal opacity, pigmentation, neovascularization, presence of ocular secretion, and conjunctival hyperemia. The following scoring system was adopted for the assessment: (0) no, (1) light, (2) moderate, and (3) severe change.Specific tests were performed at all follow-up time points. The following ophthalmic tests were adopted in the given sequence: TOT, STT-1, SMT, TBUT, fluorescein test (FT), and LGT.Tear osmolarity test was measured using an I-PEN VET osmometer . The disposable sensor chip of the device placed in contact with the conjunctiva at an angle of 45° measured the concentration of the tear solute, indicating the presence of tear film instability and aiding the qualitative diagnosis of KCS. The reference osmolarity values ranged from 296 mOsmol/L to 339 mOsmol/L .Schirmer’s tear test-1, a quantitative tear test was performed without local anesthetic. The eyes were cleaned with dry cotton and the tip of filter paper (0.5 cm long) was placed into the conjunctival sac  for 1 min. Once removed, a reading was measured immediately according to the moistened length of the filter paper. Eyes with STT-1 values <15 mm/min were considered positive for KCS .Strip meniscometry test, quantitative tear test similar to the STT was performed using a disposable paper strip  with a central column and scale printed on both sides. The strip was gently placed against the tear meniscus of each eye for a period of 5 s and the length of the central column stained blue by the tears was recorded. Values from 0 mm to 7 mm/5 s were considered positive for KCS .Tear film break-up test, a qualitative tear test, was performed by administering one drop of 1% fluorescein eye drops  on the cornea. After two blinks, the eyelid was held open manually, and the cornea was observed with a portable slit lamp (SL-15) with cobalt blue illumination, and with the assistance of a stopwatch, the tear film break-up time (appearance of the first dry spots on the cornea) was measured in seconds. Two repetitions were performed, and the average was calculated. Tear film break-up test values ≤20 s were considered positive for KCS .Fluorescein test was performed using a vital dye fluorescein that stains the corneal stroma . One droLissamine green test was performed using a vital dye, lissamine green, that stains devitalized cells in the cornea and ocular conjunctiva due to the inefficiency of tear protection caused by KCS . This teCytological analysis of third eyelid gland was performed at M0, M3, and M6, and samples were collected by fine-needle aspiration. To assemble the samples, one drop of anesthetic eye drops  was instilled to desensitize the glands of the collected eye; after exposing the third eyelid gland using tweezers, samples of the glands were collected, and one drop each of adrenaline and diclofenac sodium (Allergan) was instilled; finally, methanol and the May-Grunwald-Giemsa stain were used to fix the samples. The numbers of neutrophils, lymphocytes, and squamous cells in the epithelium were evaluated. Cell counts on each slide were performed in 10 high-power fields using an optical microscope at 400× magnification .Histopathological analysis of the conjunctiva was performed at M0 and M6 of the study. A drop of anesthetic eye drops  was instilled in each eye, and cuts were made using conjunctival scissors approximately 1.0 mm from the inferior bulbar conjunctiva and placed on a piece of paper (1 × 1 cm). Subsequently, one drop of adrenaline and one drop of diclofenac sodium (Allergan) were instilled in the conjunctiva. The collected fragments were fixed in formalin and stained using hematoxylin-eosin (HE)  and periodic acid-Schiff (PAS)  techniques. The HE-stained samples were used to evaluate the numbers of neutrophils and lymphocytes and the presence of metaplasia; the PAS technique was used to count the goblet cells. Cell count on each slide was performed in four different fields using a 400× magnification (Nikon Eclipse E200); Leica ICC50HD  was used for image recording.post hoc test was used to analyze the data from the STT-1, SMT, TBUT, TOT, and cytology and histopathological report. Friedman non-parametric test was used to compare the results of FT and LGT variables at various time points. The Kruskal–Wallis test with Dunn’s post hoc test was used to compare variables between groups. Statistical significance was set at p < 0.05. All analyses were performed using R version 4.1.3 .Bidirectional analysis of variance  for pairs of samples with Tukey’s 3; and after HPRP preparation, the final mean platelet count was 1,997,143/mm3 /mm3.The ophthalmic, cytological, and histopathological parameters of healthy dogs in the NCG are summarized in st day of treatment, making daily instillation impossible. Therefore, the dog was removed from the study and replaced by another with KCS. The dog with the adverse reaction was referred to the VH ophthalmology service, and the use of cyclosporine eye drops was recommended.The required number of applications that improved the initial ocular signs in HPRPG (n = 11 dogs/n = 22 eyes) were as follows: Four dogs/eight eyes (36% cases) received three applications, six dogs/twelve eyes (55% cases) received two applications, and one dog/two eyes (9%) received one application. Of all eyes (n = 22 eyes) that received HPRP application, only one eye (4.5%) had a mild inflammatory reaction to the blood product on the day of application, which regressed after treatment with anti-inflammatory eye drops . None of the dogs in HPRPG had to withdraw from the study. In TG, one dog/both eyes had a severe inflammatory reaction to the tacrolimus eye drops on the 1Both treatment groups showed improvements in the clinical signs and complete remission of signs at M5 . OpacifiA significant increase (p < 0.05) in the values of STT-1  and SMT The FT results are depicted in The cytology results of the third eyelid gland  are shown in The histopathological examination results of the conjunctiva are shown in This is the first study to compare the use of injectable HPRP in ambulatory care for dogs diagnosed with KCS with that of conventional topical treatment using tacrolimus 0.03% eye drops. The use of HPRP in ambulatory care requires practice and synchronization of the time between preparation and application, as the blood product must be applied instantly . ApplicaIn this study, only one eye of a dog developed a local hypersensitivity reaction caused by the application of HPRP, which was quickly treated and eventually controlled. Platelet-rich plasma from homologous and heterologous sources can cause adverse reactions owing to immunogenic differences between individual organisms. Adverse reactions to the use of autologous PRP may occur because of the substance added to the PRP preparation , 28. A bet al. [Subconjunctival application is an effective route to replace the daily instillation of eye drops . The appet al. , who caret al. . FurtherThe average weight of dogs treated with PRP was <10 kg , and somet al. [There were improvements in the clinical signs related to the ocular surface in both groups, such as decreased hyperemia, conjunctival secretion, opacity, and corneal pigmentation. Overall, TG exhibited earlier signs of remission than HPRPG. Some clinical signs showed early improvement in TG, while others stabilized only at M5. Radziejewski and Balicki  observedet al.  reportedst month of treatment and its effectiveness remained superior to HPRP throughout the study. On the SMT, despite the increase in tear production, the groups did not reach average normality. By the end of the study, HPRPG and TG showed differences in STT-1 and SMT results, but TG obtained mean STT-1 values similar to that of the NCG. The dogs in HPRPG received a maximum of three PRP applications, in contrast to the group treated with 0.03% tacrolimus that received daily instillation of eye drops twice a day throughout the study.Based on STT-1, the use of topical 0.03% tacrolimus had normalized tear production (≥15 mm/min) after the 1st month of treatment and were consistent with the results of a study involving humans, conducted by Avila et al. [The results in HPRPG were apparent after the 1a et al. , who obset al. [In a study of dogs with healthy eyes, Lamkin et al.  used theet al. –35; thuset al.  evaluateet al. [Ulcerative lesions were present in both groups at the time of diagnosis and were present up to M4 in TG and M5 in HPRPG. This result can be attributed to the fact that most treated dogs were brachycephalic dog breeds (Lhasa Apso and Shih Tzu). Their eyes are more exposed to the environment and susceptible to trauma due to their orbital structure and greater ocular prominence , 37. Faret al.  reportedThere was a considerable decrease in the total number of inflammatory cells in both groups, which was attributed to the immunomodulatory effects of the agents studied. However, the numbers of neutrophils, lymphocytes, and squamous cells in the treatment groups at the end of the study did not match the same parameters as those in the NCG. This can be explained by the chronic and progressive nature of the inflammatory process in KCS, where the objective of treatment is to reduce inflammation and improve clinical signs of the disease .et al. [In the cytopathological analysis, more neutrophils than lymphocytes were initially observed. At the final time point (M6), the neutrophil count in TG was significantly lower than in HPRPG. This can be attributed to the immunosuppressive potential of tacrolimus, which acts by calcineurin inhibition and T lymphocyte activation, thus reducing the inflammatory response . Histopaet al. , the conet al. , 40.et al. [There was also an increase in the number of goblet cells in TG and HPRPG at the end of the study. The use of tacrolimus resulted in a higher density of goblet cells in TG than in HPRPG and NCG. The action of tacrolimus on mucin-producing cells was also observed in other reports , 34. In et al. , the denTacrolimus 0.03% eye drop was found to be more efficient than HPRP in increasing tear production and the number of goblet cells. However, injectable HPRP was more efficient than tacrolimus in decreasing conjunctival inflammatory cells. Treatment with injectable HPRP requires an average of two to three applications, is safe and feasible, and can be used as a cheaper alternative or as an adjuvant to conventional treatment with topical immunosuppressants. For dogs with KCS whose owners experience difficulty in the daily use of topical eye drops or find the cost of these eye drops to be a limiting factor in the treatment, the use of injectable HPRP can be an interesting alternative.GJGE: Designed and conducted the experiment, performed all animal examinations and tests, analyzed the data, prepared the graphs, figures, and tables, and drafted the manuscript. JVGCP, LSP, and DEV: Performed all animal examinations and tests. GAN: Performed the cytological and histopathological analyses. CLS: Conceptualized the aim of the study, planned, supervised, designed, and conducted the experiment, analyzed the data, and prepared HPRP. SFA: Conceptualized the aim of the study, planned, supervised, designed, and conducted the experiment, performed all animal examinations and tests, analyzed the data, prepared the graphs, figures, and tables, and drafted the manuscript. All authors have read and approved the final manuscript.


This research intends to investigate the clinical efficacy of radiofrequency ablation and electrocautery in treating grade I or II vaginal intraepithelial neoplasia (VaIN).  This is a single-center retrospective study, which collected the clinical data of 100 patients with VaIN diagnosed by colposcopy and pathological biopsy in the Gynecology and Cervical Center of Xiangzhu Branch of the Maternal and Child Health Hospital of Guangxi Zhuang Autonomous Region between January 2020 and June 2021. Patients were divided into the study group (radiofrequency ablation treatment) and the control group (electrocautery) according to differences in treatment approaches. 6- and 12-month follow-ups were performed on all patients. Gynecological examination results, liquid-based thin-layer cytology (TCT), negative conversion of human papillomavirus (HPV), curative effects, and prognosis were recorded. P > 0.05). The analysis of postoperative follow-up complications revealed that the study group had a statistically lower overall incidence of vaginal bleeding, excessive vaginal discharge, vaginal burning sensation, and decreased vaginal elasticity than the control group (8.0% vs. 24.0% P < 0.05).  All patients completed regular follow-ups that lasted for 6 and 12 months. The 6- and 12-month cure rates of the study group were 76.0% and 92.0%, respectively, and the data in the control group were 70.0% and 82.0%, respectively. In terms of the 6- and 12-month negative conversion rates of HPV, the data in the study group were 68.0% and 78.0%, versus 60% and 68% in the control group, respectively. The lesion duration rate showed no statistical significance between the study group (8.0%) and the control group ( Both radiofrequency ablation and electrocautery have obvious clinical effects in patients with grade I or II VaIN, but the former contributed to fewer operative complications and a good prognosis, which deserves clinical promotion. Medical management is based on topical medications in the vagina, such as 5% imiquimod cream, 5-fluorouracil cream, and local estrogen. Intracavitary radiotherapy includes low- and high-dose rate brachytherapy  was the χ2 test. Statistical significance was indicated by a P value <0.05.The data were collected statistically processed by SPSS25.0 statistical software . The two independent samples P > 0.05). Among the 50 patients in the study group, 42 patients were diagnosed as stage I VaIN, including 39 with hrHPV positive and 30 patients with I-II CIN; the results of TCT showed 13 cases of NILM, 12 cases of ASCUS, 20 cases of LSIL, and 5 cases of HSIL. While in the control group, 34 patients were HPV positive and 28 patients were complicated with I-II CIN; the results of TCT showed NILM, ASCUS, LSIL, and HSIL in 15, 15, 16, and 4 cases, respectively.The two groups showed similar clinical characteristics . PatientP > 0.05)  .P > 0.05)  .P < 0.05)  in the radiofrequency ablation group and 24.0% (12/50) in the electrocautery group, showing no significant difference ( < 0.05) . At the VaIN is a precancerous lesion composed of a typical hyperplastic squamous cell groups, including atypical hyperplasia and carcinoma in situ, which can be classified as grades I-III VaIN according to the degree of atypical hyperplasia. Studies have reported that low-grade VAIN has a tendency to self-heal, but not all can be reversed with some of the lesions persist or even progress . Risk fa2 laser ablation and photodynamic therapy in the treatment of VaIN. For example, Han et al. [2 laser to treat VaIN patients. Most patients need repeated laser treatment. For instance, CO2 laser ablation has a certain residual rate, which increases the possibility of repeated operations, resulting in an increased risk of postoperative vaginal stricture, shortening, scar formation, and difficulty in sexual intercourse, as well as the need for long-term follow-up. It can be seen that each treatment has its advantages and disadvantages. In the control group, electrocautery was performed to scald the vaginal lesions, and the thermal effect was used to carbonize the tissue protein and occlude the blood vessels; thus, inducing the atrophy and even necrosis and shedding of the diseased wound tissue. Moreover, due to the mild and uniform heating during electrocautery, the action point is relatively accurate, and the depth and treatment can be judged according to the changes in the color of the wound surface, with high safety. It has been reported that the cure rate of electrocautery for VaIN is 25%–88% [The current treatment of VaIN is roughly the same as CIN, with fewer adverse reactions, which is suitable for young and mild patients; however, the long treatment course leads to poor patient compliance, resulting in low stability of treatment efficacy, while surgical treatment is traumatic and requires high requirements for operators, with postoperative complications that have a certain impact on the quality of life of patients. Therefore, choosing an appropriate treatment is of utmost importance. In recent years, a number of studies have reported better clinical effects of COn et al.  used phon et al.  reported 25%–88% . In this 25%–88% . In thisP > 0.05). The increase in the negative conversion rate of HPV during the follow-up period also indicates the long-term effect of radiofrequency ablation. While resolving the lesion, the treatment can lower the viral load to some certain extent and even eliminate HPV infection. The results showed that the negative conversion rate of other hrHPV was higher than that of HPV16/18.In addition, the negative conversion rates of HPV in the radiofrequency ablation group and electrocautery group in 6 months were 68% and 60%, respectively, and those in 12 months were 78% and 68%, respectively, showing no statistical significance (In order to better treat VaIN and reduce the occurrence of complications after physiotherapy, all patients with VaIN were treated with Baofukang suppositories postoperatively. As a traditional Chinese medicine suppository, the Baofukang suppository is mainly composed of zedoary turmeric oil and borneol. Zedoary turmeric can activate blood to remove blood stasis, relieve pain, and remove saprophytic muscle; while borneol has the effects of inducing resuscitation, alleviating swelling and pain, removing saprophytic muscles, cooling blood, and relieving itching . The appThrough the analysis of the curative effect of radiofrequency ablation in patients with VaIN, this study found that radiofrequency ablation was effective in patients with stages I-II VaIN, which could not only accurately locate the target tissue of injury but also reduce the damage to the surrounding normal tissue. Postoperative treatment with Baofukang suppositories is conducive to accelerating wound healing, alleviating vaginal scar hyperplasia and tissue adhesion, and reducing the symptoms of postoperative vaginal bleeding and vaginal discharge. The vaginal anatomy and function remained good after treatment, but a small number of patients in the electrocautery group developed complications such as vaginal burning sensation, decreased vaginal elasticity, and stricture, which affect their quality of life and their satisfaction with subsequent colposcopy.To sum up, both radiofrequency ablation and electrocautery in combination with Baofukang suppositories are effective in the treatment of VaIN, which can promote lesion healing, reduce recurrence, and improve the cure rate of patients. But relatively speaking, radiofrequency ablation has higher safety, which contributes to fewer postoperative complications and a higher cure rate. Thus, radiofrequency ablation combined with Baofukang suppositories can be given priority in the treatment of stages I-II VaIN patients. However, there is still some room for improvement in this study. The sample size of this study may be too small to detect a difference between the two approaches. Consequently, a multicenter, large sample, and long-term follow-up study is needed to confirm the research findings.