Delivery of WeChat-Based HIV Result e-Reports in Social Networks for Recruitment of High-Risk Population: Baseline Data From a Cluster Randomized Controlled Trial.

BACKGROUND: Disclosure of infectious disease status to social network peers can facilitate reaching and early detection among high-risk populations. In this era of social media, globally, HIV/AIDS represents a high burden of infectious disease. Thus, delivery of an HIV result e-report via social media presents a new approach that has the potential to improve contact with and enrollment of the high-risk population in research studies and routine practice. OBJECTIVE: This study explores the effectiveness and associated factors of a recruitment strategy (ie, WeChat-based HIV e-report delivery in social networks) on the enrollment of men who have sex with men (MSM) for an HIV testing intervention study. METHODS: This was an enrollment result analysis of an ongoing cluster randomized controlled trial (RCT) aiming to promote HIV testing among MSM. Recruitment of potential participants was based on the unit of an egocentric social network, which includes 1 core member (an offline tested ego as the recruiter) and several network members (online alters as network associates). Alters' enrollment and alters' transformation to ego-recruiters (alter-ego) were measured as outcomes. Recruitment outcomes were compared between the exchangeable and regular e-report groups of the RCT. Associated factors of both outcomes were also investigated, including sociodemographic characteristics, health behaviors, social network characteristics, e-report types, and online delivery information. Binary outcomes were modeled using logistic models, with Firth correction for rare events. Qualitative interviews were conducted to understand facilitators and barriers in detail for alter-ego as the subsequent wave's recruiter. RESULTS: The e-report of 1157 egos who tested offline were delivered to 5165 alters in 3 recruitment waves; eventually, 1162 eligible alters enrolled in this RCT (response rate: 22.5%). In the exchangeable e-report group, 544 egos recruited 467 alters, of which 35 alters transformed to alter-egos (7.5%), whereas in the regular e-report group, 613 egos recruited 695 alters, of which 40 alters transformed to alter-egos (5.8%). Alters' enrollment at first wave was associated with a higher number of e-reports being forwarded by the egos. Alters' transformation to alter-egos for the subsequent wave was associated with the exchangeable e-report, higher income, being a Guangzhou resident, unprotected anal intercourse, preferring self-testing, and viewing senders' e-reports frequently. Qualitative interviews revealed that the lack of awareness of e-reports' function and inadequate access to e-reports at offline testing facilities were major barriers to alters' transformation to offline ego-recruiters. CONCLUSIONS: The delivery of e-report was feasible in MSM social network, and the success and sustainability of online recruitment depended on high levels of familiarity among MSM with the digital tool. The HIV e-report exchange mechanism might promote MSM to test HIV offline to get their own e-report for exchange in the community. The e-report provides an innovative recruitment method with great potential to trace direct contacts for infectious diseases studies.

WeChat-Based HIV e-Report, a New Approach for HIV Serostatus Requests and Disclosures Among Men Who Have Sex With Men: Prospective Subgroup Analysis of a Randomized Controlled Trial

BACKGROUND: Requesting and disclosing HIV serostatus is associated with a reduction in HIV transmission among men who have sex with men (MSM). However, the reliability of common methods for HIV serostatus request and disclosure is inadequate. Validated approaches for requesting and disclosing HIV serostatus are necessary. OBJECTIVE: The objective of this study was to investigate the use of the HIV e-report as authentic evidence of HIV serostatus among the MSM community in Guangzhou, China. Additionally, the study aimed to explore its correlation with HIV serostatus requesting and disclosure receiving behavior. METHODS: This study is a subgroup analysis of a cluster randomized controlled trial (RCT) that enrolled 357 participants during the first year. Participants in this RCT were recruited from the WeChat-based HIV testing service miniprogram developed by Guangzhou Center for Disease Control and Prevention, China. Participants completed web-based questionnaires at baseline and at the month 3 follow-up, which covered sociodemographic characteristics, HIV-related information, HIV serostatus requests, receiving HIV serostatus disclosures, and HIV e-report usage. Univariate and multivariate logistic regressions were used for data analysis. RESULTS: The WeChat-based HIV e-report was available in Guangzhou when the RCT project started. At the month 3 follow-up, 32.2% (115/357) of participants had their own HIV e-reports, and 37.8% (135/357) of them had received others' HIV e-reports. In all, 13.1% (27/205) and 10.5% (16/153) of participants started to use HIV e-reports to request the HIV serostatus from regular and casual male sex partners, respectively. Moreover, 27.3% (42/154) and 16.5% (18/109) of the regular and casual male sex partners, respectively, chose HIV e-reports to disclose their HIV serostatus. Compared to MSM who did not have HIV e-reports, those who had HIV e-reports and stated, "I had had my own HIV e-report(s) but hadn't sent to others" (multivariate odds ratio 2.71, 95% CI 1.19-6.86; P=.02) and "I had had my own HIV e-reports and had sent to others" (multivariate odds ratio 2.67, 95% CI 1.07-7.73; P=.048) were more likely to request HIV serostatus from their partners. However, no factor was associated with receiving an HIV serostatus disclosure from partners. CONCLUSIONS: The HIV e-report has been accepted by the MSM community in Guangzhou and could be applied as a new optional approach for HIV serostatus requests and disclosures. This innovative intervention could be effective in promoting infectious disease serostatus disclosure among the related high-risk population. TRIAL REGISTRATION: ClinicalTrials.gov NCT03984136; https://clinicaltrials.gov/show/NCT03984136. INTERNATIONAL REGISTERED REPORT IDENTIFIER (IRRID): RR2-10.1186/s12879-021-06484-y.

Correction: WeChat-Based HIV e-Report, a New Approach for HIV Serostatus Requests and Disclosures Among Men Who Have Sex With Men: Prospective Subgroup Analysis of a Randomized Controlled Trial.

[This corrects the article DOI: 10.2196/44513.].

The N-terminal polypeptide derived from vMIP-II exerts its antitumor activity in human breast cancer through CXCR4/miR-7-5p/Skp2 pathway.

Breast cancer is a malignant tumor with the highest incidence in women of the world. CXCR4 and Skp2 are highly expressed in breast cancer cells and CXCR4 was positively correlated with Skp2 by interference or overexpression. The microRNA array was used to detect the differentially expressed spectrum of micro RNAs in breast cancer cells the changes of miR-7-5p after CXCR4 inhibitor (NT21MP) treatment to block the CXCR4/SDF-1 pathway was founded. MiR-7-5p has been found to be correlated with Skp2 in various tumors in the literature, and Skp2 expression can be regulated by transfection with miR-7-5p mimics or inhibitors. The expression level of miR-7-5p was upregulated or downregulated after CXCR4 interference or overexpression. Combined with the correlation between CXCR4 and miR-7-5p in the chip results, CXCR4 may regulate Skp2 through miR-7-5p. Epithelial cells have the morphological characteristics of mesenchymal cells for some reason called epithelial-mesenchymal transformation (EMT). Transfection of miR-7-5p mimics into drug-resistant cells reduced Skp2 levels, decreased the expression of Vimentin, Snail, and slug, and increased the expression of E-cadherin. CXCR4 inhibitor (NT21MP) can reverse the EMT changes caused by miR-7-5p inhibitor. Similarly, in vivo results suggesting that CXCR4 inhibitors can reverse the EMT phenotype of drug-resistant breast cancer cells through the CXCR4/miR-7-5p/Skp2 pathway. In summary, the CXCR4/miR-7-5p/Skp2 signaling pathway plays an important role in the progression of breast cancer. This study provides a theoretical basis for the treatment of breast cancer by targeting the CXCR4 pathway.

Long noncoding RNA H19 acts as a miR-340-3p sponge to promote epithelial-mesenchymal transition by regulating YWHAZ expression in paclitaxel-resistant breast cancer cells.

Breast cancer (BC) is the leading cause of cancer-related death in women worldwide and one of the most prevalent malignancy. In recent years, increasing evidence had illuminated that long noncoding RNAs (lncRNAs) serve as critical factors in multiple tumor progression, including BC. Emerging references had indicated that the lncRNA H19 acts as significant roles in tumor progression and epithelial-mesenchymal transition (EMT). However, the underlying molecular mechanisms and biological roles of H19 in BC invasion, metastasis and EMT are still unclear. In this study, it was detected that the expression level of H19 was increased in BC paclitaxel-resistant (PR) cells subline (MCF-7/PR) in comparison with MCF-7 parental cells. In vitro, there were demonstrated that H19 overexpression promoted BC cells proliferation, metastasis, invasion and EMT procedures, and suppressed cells apoptosis. Whereas, H19 suppression resulted in the contrary biological effects. Besides, bioinformatics tools and dual-luciferase reporters assays indicated that miR-340-3p could act as a potential target gene of H19, the underlying mechanism studies proved that H19 could act as a competing endogenous RNA (ceRNA) via competitively binding miR-340-3p to promote BC cell proliferation, metastasis and EMT by regulating tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein zeta (YWHAZ) and potentiate the Wnt/ß-catenin signaling in BC cells. In summary, our findings demonstrated that H19 could act as a ceRNA in BC progression, metastasis and EMT through modulating miR-340-3p/YWHAZ axis and activating the canonical Wnt/ß-catenin signaling pathway, indicating that H19 might act as an underlying therapeutic target and prognostic biomarker for BC therapy.

Correction: Associations Between Affective States and Sexual and Health Status Among Men Who Have Sex With Men in China: Exploratory Study Using Social Media Data.

[This corrects the article DOI: 10.2196/13201.].

Associations Between Affective States and Sexual and Health Status Among Men Who Have Sex With Men in China: Exploratory Study Using Social Media Data.

BACKGROUND: Affective states, including sentiment and emotion, are critical determinants of health. However, few studies among men who have sex with men (MSM) have examined sentiment and emotion specifically using real-time social media technologies. Moreover, the explorations on their associations with sexual and health status among MSM are limited. OBJECTIVE: This study aimed to understand and examine the associations of affective states with sexual behaviors and health status among MSM using public data from the Blued (Blued International Inc) app. METHODS: A total of 843,745 public postings of 377,610 MSM users located in Guangdong were saved from the Blued app by automatic screen capture. Positive affect, negative affect, sexual behaviors, and health status were measured using the Simplified Chinese Linguistic Inquiry and Word Count. Emotions, including joy, sadness, anger, fear, and disgust, were measured using the Weibo Basic Mood Lexicon. A positive sentiment score and a positive emotion score were also calculated. Univariate and multivariate linear regression models on the basis of a permutation test were used to assess the associations of affective states with sexual behaviors and health status. RESULTS: A total of 5871 active MSM users and their 477,374 postings were finally selected. Both positive affect and positive emotions (eg, joy) peaked between 7 AM and 9 AM. Negative affect and negative emotions (eg, sadness and disgust) peaked between 2 AM and 4 AM. During that time, 25.1% (97/387) of negative postings were related to health and 13.4% (52/387) of negative postings were related to seeking social support. A multivariate analysis showed that the MSM who were more likely to post sexual behaviors were more likely to express positive affect (beta=0.3107; P<.001) and positive emotions (joy: beta=0.027; P<.001), as well as negative emotions (sadness: beta=0.0443; P<.001 and disgust: beta=0.0256; P<.001). They also had a higher positive sentiment score (beta=0.2947; P<.001) and a higher positive emotion score (beta=0.1612; P<.001). The MSM who were more likely to post their health status were more likely to express negative affect (beta=0.8088; P<.001) and negative emotions, including sadness (beta=0.0705; P<.001), anger (beta=0.0058; P<.001), fear (beta=0.0052; P<.001), and disgust (beta=0.3065; P<.001), and less likely to express positive affect (beta=-0.0224; P=.02). In addition, they had a lower positive sentiment score (beta=-0.8306; P<.001) and a lower positive emotion score (beta=-0.3743; P<.001). CONCLUSIONS: The MSM social media community mainly expressed their positive affect in the early morning and negative affect after midnight. Positive affective states were associated with being sexually active, whereas negative affective states were associated with health problems, mostly about mental health. Our finding suggests the potential to deliver different health-related intervention strategies (eg, psychological counseling and safe sex promotion) on a social media app according to the affective states of MSM in real time.

sRAGE plays a role as a protective factor in the development of PCOS by inhibiting inflammation.

This study determined the effect of exogenous soluble receptor for advanced glycation end products (sRAGE) on the pro-inflammatory activities that occur during polycystic ovary syndrome (PCOS) in human follicular cells and explored a potential mechanism for preventing the development of inflammation. Follicular fluid was allocated into one of three treatment groups (0, 0.6, and 1.2 µg mL-1 of sRAGE). Collectively, these results indicate that exogenous sRAGE supplementation alleviates inflammation in ovarian follicular granulosa cells by regulating p-ERK and AP-1 signaling.

Long noncoding RNA AC073284.4 suppresses epithelial-mesenchymal transition by sponging miR-18b-5p in paclitaxel-resistant breast cancer cells.

Breast cancer (BC) is the most prevalent malignant cancer in the world, is the leading cause of cancer-related death female. Recently, there is accumulating evidence that long noncoding RNAs (lncRNAs) might as an important role in the progression of BC. (epithelial-mesenchymal transition (EMT) is considered to play a vital role in tumor cells migration and invasion. Nevertheless, the entire biological mechanisms and functions of lncRNAs in tumor migration, invasion, and EMT remain uncertain. In the present research, we observed that the expression of lncRNA AC073284.4 was downregulated in BC paclitaxel-resistant (PR) cells (MCF-7/PR) and tissues. Bioinformatics analysis predicted that miR-18b-5p was a direct target of AC073284.4, which has been validated by dual-luciferase reporter gene assay. We further proved that AC073284.4 could directly bind to miR-18b-5p and relieve the suppression for dedicator of cytokinesis protein 4 (DOCK4). Furthermore, the underlying functional experiments demonstrated that AC073284.4 might sponge miR-18b-5p to attenuate the invasion, metastasis, and EMT of BC cell through upregulating DOCK4 expression. In summary, AC073284.4 might serve as a competing endogenous RNA (ceRNA) in BC progression via modulating miR-18b-5p/DOCK4 axis, which weakens EMT and migration of BC. These results suggesting that AC073284.4 might function as a potential novel diagnostic biomarker in the progression of BC.

Metformin Regulates Key MicroRNAs to Improve Endometrial Receptivity Through Increasing Implantation Marker Gene Expression in Patients with PCOS Undergoing IVF/ICSI.

To some extent, the use of metformin may improve endometrial receptivity and pregnancy outcomes of women with polycystic ovarian syndrome (PCOS) undergoing in vitro fertilization/intracytoplasmic sperm injection. However, the mechanism is not well-known. The endometrium of metformin-treated group (metformin-treated patients with PCOS) and the control group (non-metformin-treated patients with PCOS) were analyzed for the expression of homeobox A10 (HOXA10) and integrin beta-3 (ITGB3) and differential micro RNA (miRNA) expression profiles. On this basis, miRDB and Target Scan databases were used to predict and screen out that miR-491-3p and miR-1910-3p may target HOXA10 and ITGB3. Furthermore, we verified the effects of metformin on the expression of HOXA10 and ITGB3, and regulatory effects of miR-1910-3p and miR-491-3p on HOXA10 and ITGB3 using Ishikawa cell line. Metformin induced a significant dose-dependent upregulation of HOXA10 and ITGB3. The results from the microarray analyses showed there were 40 differentially expressed miRNAs between the 2 groups. Among them, miR-1910-3p and miR-491-3p were the 2 significantly downregulated miRNAs. Bioinformatics prediction indicated that HOXA10 and ITGB3 are potential target genes for miR-1910-3p and miR-491-3p. In Ishikawa cells transfected with miR-491-3p mimics, the expression of HOXA10 and ITGB3 on both messenger RNA (mRNA) and protein level were lower than those in control group (P < .001). Also, the expression of HOXA10 mRNA and protein was lower in Ishikawa cells transfected with miR-1910-3p mimics (P < .001). However, no significant changes in ITGB3 levels were observed in cells transfected with miR-1910-3p mimics (P > .05). Metformin likely improves endometrial receptivity through downregulating the expression of miR-491-3p and miR-1910-3p, thereby increasing the expression of HOXA10 and ITGB3 in the endometrium of PCOS women.

The N-terminal polypeptide derived from viral macrophage inflammatory protein II reverses breast cancer epithelial-to-mesenchymal transition via a PDGFRα-dependent mechanism.

NT21MP, a 21-residue peptide derived from the viral macrophage inflammatory protein II, competed effectively with the natural ligand of CXC chemokine receptor 4 (CXCR4), stromal cell-derived factor 1-alpha, to induce apoptosis and inhibit growth in breast cancer. Its role in tumor epithelial-to-mesenchymal transition (EMT) regulation remains unknown. In this study, we evaluated the reversal of EMT upon NT21MP treatment and examined its role in the inhibition of EMT in breast cancer. The parental cells of breast cancer (SKBR-3 and MCF-7) and paclitaxel-resistant (SKBR-3 PR and MCF-7 PR) cells were studied in vitro and in combined immunodeficient mice. The mice injected with SKBR-3 PR cells were treated with NT21MP through the tail vein or intraperitoneally with paclitaxel or saline. Sections from tumors were evaluated for tumor weight and EMT markers based on Western blot. In vitro, the effects of NT21MP, CXCR4 and PDGFRα on tumor EMT were assessed by relative quantitative real-time reverse transcription-polymerase chain reaction, western blot and biological activity in breast cancer cell lines expressing high or low levels of CXCR4. Our results illustrated that NT21MP could reverse the phenotype of EMT in paclitaxel-resistant cells. Furthermore, we found that NT21MP governed PR-mediated EMT partly due to controlling platelet-derived growth factors A and B (PDGFA and PDGFB) and their receptor (PDGFRα). More importantly, NT21MP down-regulated AKT and ERK1/2 activity, which were activated by PDGFRα, and eventually reversed the EMT. Together, these results indicated that CXCR4 overexpression drives acquired paclitaxel resistance, partly by activating the PDGFA and PDGFB/PDGFRα autocrine signaling loops that activate AKT and ERK1/2. Inhibition of the oncogenic EMT process by targeting CXCR4/PDGFRα-mediated pathways using NT21MP may provide a novel therapeutic approach towards breast cancer.

Correlation analysis between ApoM gene-promoter polymorphisms and coronary heart disease.

OBJECTIVES: Apolipoprotein M (ApoM), a 25-kDa plasma protein belonging to the lipocalin protein family, is predominantly associated with high-density lipoprotein cholesterol (HDL-C). Studies have suggested ApoM to be important for the formation of pre-ß-HDL and to increase cholesterol efflux from macrophage foam cells. The aim of this study was to explore the association of single-nucleotide polymorphisms (SNPs) in the ApoM promoter with coronary atherosclerotic disease (CAD), and the contribution of latent factors. METHODS: ApoM was measured in samples from two separate case-control studies, of whom 88 patients developed CAD and 88 were controls. Whole-blood samples from subjects were genotyped by PCR-restriction fragment length polymorphism (PCR-RFLP). Luciferase activities were measured for HepG2 cells with two SNPs, rs805296 (T-778C) and rs940494 (T-855C), and after interfering with or overexpressing the predicted transcription factors. The ability of the SNPs to combine with nucleoproteins was analysed by electrophoretic mobility shift assay (EMSA). RESULTS: Mean plasma ApoM concentrations in the CAD and non-CAD groups were 9.58 ± 4.30 and 12.22 ± 6.59 µg/ ml, respectively. Correlation studies of ApoM concentrations with several analytes showed a marked positive correlation with HDL-C, fasting plasma glucose and triglyceride levels. The CC genotype showed lower luciferase activities compared to the TC and TT genotypes. The ApoM-855 mutant-type could bind to the AP-2α. Interference and overexpression of AP-2 increased and decreased luciferase activities of the wild and mutant types to different degrees. CONCLUSION: ApoM may be a biomarker of CAD. ApoM-855 T→C substitution provides binding sites for AP-2α and reduces ApoM transcription activity.

The effects of anticancer drugs TSA and GSK on spermatogenesis in male mice.

OBJECTIVE: The effect of anticancer drugs Trichostation A (TSA) and GSK2126458 (GSK) on genetic recombination of sperm meiosis in mice was investigated, and their clinical feasibility of fertility preservation in cancer patients was also assessed. METHODS: Eighteen Kunming mice were randomly given TSA or GSK at the concentrations of 0, 0.1 and 0.2 umol/L for three months. Immunofluorescence was used to evaluate the genetic recombination of homologous chromosomes and fidelity of chromosome synapsis. Sperm density, motility and viability were also examined to investigate the spermatogenic function. RESULTS: The average number of MLH1 foci in each spermatocyte was greatly higher in TSA (0.1) group than that in control (P<0.05), but no difference with the TSA (0.2) group (P>0.05). The frequency of SC with no MLH1 foci was lower while the frequency of SC with one MLH1 foci was higher in spermatocyte of mice with different doses of TSA compared with controls (P<0.05). The weight of left testis in TSA (0.1) group was significant decreased compared with that in control (P<0.05). However, no significant differences were observed in average number of MLH1, frequency of SC with 0-3 MLH1 foci, spermatocyte percentage of XY chromosomes containing MLH1 foci and percentages of cells containing gaps and splits among groups with or without the treatment of GSK. Furthermore, there were no statistical differences in body weight, testicular weight, sperm density, sperm motility and sperm viability among the three groups. CONCLUSION: TSA increased genetic recombination frequency of spermatocyte meiosis. GSK had no significant effect on genetic recombination frequency of spermatocyte meiosis and spermatogenic function.

[Effects of miRNA-21 on paclitaxel-resistance in human breast cancer cells].

OBJECTIVE: To investigate the effects of miR-21 on paclitaxel-resistance in human breast cancer MCF-7/PR and SKBR-3/PR cells. METHODS: Paclitaxel-resistant human breast cancer cell lines MCF-7/PR and SKBR-3/PR were established by stepwise selection in increasing concentration of paclitaxel. Cellular morphology, mRNA and protein level of MDR1, BCRP and MRP1 in MCF-7/PR and SKBR-3/PR cells were determined. The expression of Bax, Bcl-2 and miR-21 in parental and paclitaxel-resistant cells was detected by RT-PCR and Western blotting. The synthetic miR-21 inhibitor or miR-21 mimic were transfected into MCF-7/PR, SKBR-3/PR and MCF-7, SKBR-3 cells with Lipofectamine 2000. The miR-21 levels were determined by RT-PCR, and P-gp, Bcl-2 and Bax protein levels were examined by Western blotting. MTT assay was used to measure the cell viability, and flow cytometry was performed to analyze the cell cycle and apoptosis. RESULTS: The levels of MDR1, BCRP, MRP1, Bcl-2/Bax and miR-21 in MCF-7/PR and SKBR-3/PR cells were significantly higher than those in MCF-7 and SKBR-3 cells. The protein levels of P-gp, Bcl-2 were up-regulated, and Bax was down-regulated compared with parental cells. MiR-21 was significantly down-regulated after miR-21 inhibitor was transfected; and the levels of MDR1, BCRP, MRP1 and Bcl-2/Bax (P <0.05) were also down-regulated. MiR-21 inhibitors significantly suppressed G0/G1 transition of the cell cycle, and induced cell apoptosis in MCF-7/PR and SKBR-3/PR cells. MTT results showed that miR-21 inhibitors induced sensitivity of MCF-7/PR and SKBR-3/PR cells to paclitaxel. And miR-21 mimic can increase the expression of MDR1, Bcl-2/Bax and change cell morphology from parental cells to resistant cells. RESULTS: The established MCF-7/PR and SKBR-3/PR breast cancer cells show typical multidrug resistance characteristics, which can be used as the model for drug resistance study. Down-regulated miR-21 expression in MCF-7/PR and SKBR-3/PR breast cancer cells can enhance cell sensitivity to paclitaxel.

[A Method Research on Environmental Damage Assessment of a Truck Rollover Pollution Incident].

With high occurrence of sudden water pollution incident, China faces an increasingly severe situation of water environment. In order to deter the acts of environmental pollution, ensure the damaged resources of environment can be restored and compensated, it is very critical to quantify the economic losses caused by the sudden water pollution incident. This paper took truck rollover pollution incidents in Chongqing for an example, established a set of evaluation method for quantifying the environmental damage, and then assessed the environmental damage by the method from four aspects, including the property damage, ecological environment and resources damages, the costs of administrative affairs in emergency disposal, and the costs of investigation and evaluation.

[Construction and identification of anti-HER2 phage display single chain fragment of variable region library in human breast cancer].

OBJECTIVE: To construct human phage single-chain antibody (scFv) library against breast cancer, and to identify anti-HER2 specific antibodies from the human phage display scFv library to offer a stronger affinity sequence targeting HER2 for fusion protein targeting HER2 and CXCR4. METHODS: Total RNA was extracted from the adjacent lymphatic tissue harvested from breast cancer patients. The variable regions of the whole antibody were amplified by using RT-PCR and were cloned into the vector pCANTAB-5E through a linker. The products were electroporated into competent E.coli TG1 cells. Recombinant phages specific for breast cancer cells were enriched in SKBR-3 after four rounds. The antigen-positive clones were selected by ELISA and immunohistochemistry. RESULTS: The fragment of VH and VL were about 375 and 330 bp and were linked in vitro to form scFv of 750 bp that was resistant to the breast cancer HER2 single strand. A fusion phage display library that contained total of 2.48×10(8) pfu /ml was established. ELISA and immunohistochemical results confirmed that the antibody has a strong affinity with HER2 antigen in breast cancer tissue. Compared to human IgG antibody, a scFv phage library against human breast cancer was successfully constructed with high capacity. The scFv was highly specific to HER2 antigen and the sequencing results indicated that VL and VH genes were highly homologous with the variable region of human antibody. CONCLUSION: This strategy may achieve new targeted antibody resistant to the breast cancer for clinical treatment and provide a carrier that uses HER2 as a target of the fusion protein for anti-tumor therapy.

[Influence of recombination abnormality in spermatocyte meiosis on the pathology of patients with non-obstructive azoospermia].

OBJECTIVE: To primarily study the influence of recombination abnormality in human spermatocyte meiosis on the pathology of the patients with non-obstructive azoospermia (NOA). METHODS: We obtained testis tissues from 6 NOA patients by testicular biopsy and divided the tissue of each patient into 2 portions, one for pathological examination and the other for immunofluorescent staining. We observed the synaptonemal complex and the numbers of the recombination sites on homologous chromosomes, and analyzed the relationship between abnormal recombination and pathological findings. RESULTS: Pathological examination showed that the basement membrane of the seminiferous tubules was thickened in 3 of the cases and atrophied in the other 3, the number of autosomal MLH1 foci in a spermatocyte ranging from 10 to 50 in the former 3, and from 30 to 50 in the latter 3. CONCLUSION: The increased range of the homologous chromosomal recombination frequency may be one of the possible factors for the thickening of seminiferous tubule basement membrane and even lumen occlusion in NOA patients.

[Construction of pBIFC-VN173-CXCR4 and pBIFC-VC155-NT21MP eukaryotic expression plasmids and their interaction in living cells].

OBJECTIVE: To construct pBIFC-VN173-CXCR4 and pBIFC-VC155-NT21MP eukaryotic expression plasmids and to investigate the interaction of chemokine receptor 4 (CXCR4) and viral macrophage inflammatory protein-II(vMIP-II) N terminal 21 peptides (NT21MP) in living cells. METHODS: DNA fragment encoding NT21MP was chemically synthesized and inserted into BiFC eukaryotic expression vector pBIFC-VC155. The full length of CXCR4 DNA fragment was amplified by RT-PCR from SKBR (3) cells and inserted into BiFC eukaryotic expression plasmid pBIFC-VN173. Two recombinant vectors were identified by restriction enzyme digestion and DNA sequencing. The recombinant vectors were cotransfected into Africa green monkey kidney fibroblast COS-7 cells by using Lipofectamine 2000. The interaction of NT21MP and CXCR4 was detected by bimolecular fluorescence complementation (BiFC) assay. RESULTS: The restriction enzyme digestion and DNA sequences and open read frames of two vectors were consistent with experiment design. The BiFC plasmids were successfully cotransfected into the target cells and expressed. The strong BiFC signals were detected in pBIFC-VN173-CXCR4 and pBIFC-VC155-NT21MP cotransfected cells and the fluorescence signal was located in the cytoplasm. CONCLUSION: The eukaryotic expression plasmids for BiFC assay are successfully constructed. The interaction of NT21MP and CXCR4 in living cells can be detected by using this technology.

[Abnormalities of meiotic recombination in Han Chinese azoospermic patients].

OBJECTIVE: To analyze defective homologous chromosomal recombination in Han Chinese azoospermic patients. METHODS: Testicular biopsy samples from 7 healthy controls and 7 Han Chinese azoospermic patients including 2 obstructive azoospermia (OA group) and 5 non-obstructive azoospermia (NOA group) were analyzed. Immunofluorescence staining was performed to categorize early stage cells at meiosis prophase and to analyze chromosome pairing and recombination of pachytene spermatocyte. Newly developed meiotic proteins antibodies (anti-SCP3, anti-synaptonemal complex proteins 3, anti-MLH1, anti-Mut-L Homolog 1, anti-CREST, chromosome centromere antibody) were used to identify synaptonemal complex (anti-SCP3), recombination sites (anti-MLH1) and centromere (anti-CREST), respectively. Staging of spermatocyte was determined according to SCP3 formation progression. Qualitative data were compared by a Chi-square test, and ANOVA was used to analyze quantitative data. RESULTS: Respectively, 2346 and 2932 spermatocytes were categorized in the controls and azoospermic patients. The proportions of zygotene cells in both OA group and NOA group were significantly higher than that of the control group. Investigation of 1967 pachytene cells from the controls and 354 pachytene cells from azoospermic patients indicated that the mean MLH1 foci per pachytene cell of NOA group was statistically lower than that of the controls. Compared with the controls, incomplete synaptonemal complexes cells (containing gap and/or split) were significantly increased in the NOA group. CONCLUSION: Delayed meiosis prophase is relatively common in azoospermic patients, and changes in quantity and distribution of recombination foci may be the cause for spermatogenesis arrest in Han Chinese population.