[HCV and Treponema pallidum infection status in HIV/AIDS cases in Yunnan province, January-June, 2020].

Objective: To understand the infection status of HCV and Treponema pallidum (TP) in HIV/AIDS cases in Yunnan province,and identify the risk factors. Methods: Between January 1 and June 30 in 2020,a cross-sectional survey was conducted in Yunnan. Two enzyme-linked immunosorbent assay (ELISA) kits were used to detect anti-HCV, the positive results of both two kits indicated HCV infection. ELISA and syphilis toluidine red untreated serum test were applied to identify TP infection. Both Excel 2016 and SPSS 22.0 software were used for statistical analysis, and logistic regression model was conducted to identify the relevant factors of HCV and TP infection. Results: A total of 5 922 HIV/AIDS cases were included in this study, the infection rates of HCV and TP were 6.5% (383/5 922) and 5.8% (344/5 922) respectively. The co-infection rate of HCV and TP was 0.4% (22/5 922). The risk for HCV infection in HIV/AIDS cases was higher in younger age groups compared with age group ≥50 years (15-19:aOR=3.53;20-29:aOR=3.02;30-39:aOR=2.91;40-49:aOR=3.61), in males than in females (aOR=2.31), in the married and unmarried than in the divorced or widowed (married:aOR=1.61;unmarried:aOR=1.63), in other ethnic groups than in Han ethnic group (aOR=1.70), in people with lower education level than in people with education level of college and above (primary school degree and below:aOR=4.69;middle school:aOR=3.96), in people living in the central and western Yunnan than in people living in eastern Yunnan (central Yunnan:aOR=2.46; western Yunnan:aOR=7.08), in injection drug users than in MSM (aOR=131.08). The risk of TP infection in HIV/AIDS cases was higher in people with education level of college and primary school than in middle school degree (primary school and below:aOR=1.73;college and above:aOR=1.77), in people with other occupations than in farmers (aOR=1.39), in people living in eastern Yunnan than in people living in western Yunnan (aOR=1.75); in MSM than in people with heterosex (aOR=9.75). Conclusions: A certain proportion of HIV/AIDS cases reported between January and June in 2020 in Yunnan were co-infected with HCV and TP, many factors were associated with the co-infection. It is suggested to strengthen HCV and TP tests in HIV/AIDS cases and conduct active treatment of the co-infection.

[Performance of limiting-antigen avidity enzyme immunoassay and pooling PCR in detection of recent HIV-1 infection among men who have sex with men in Yunnan province].

Objective: To assess and compare the performance of limiting-antigen avidity enzyme immunoassay (LAg-Avidity EIA) and pooling PCR in the surveillance for recent infection rates of HIV-1 in men who have sex with men (MSM). Methods: Blood samples were collected from MSM selected through snowball sampling method in sentinel surveillance in 13 prefectures of Yunnan province from 2016 to 2017. The samples were tested for HIV-1 antibody. The confirmed positive samples were tested by LAg-Avidity EIA. The negative samples were tested by pooling PCR. The recent infection rates of HIV-1 were estimated by the algorithm based on LAg-Avidity EIA and pooling PCR respectively. The two results were compared. Results: During 2016-2017, a total of 5 363 blood samples were collected from MSM, in which 407 samples were HIV-1 positive (including 177 positive tested previously) and 4 956 samples were HIV-1 negative. A total of 211 samples(91.7%) were tested by LAg-Avidity EIA, 69 were confirmed to be recent infections. A total of 4 469 samples were tested by pooling PCR, 8 were confirmed to be acute infections. The recent infection rates of HIV-1 from 2016 to 2017 estimated by LAg-Avidity EIA were 3.36% and 4.84%, and the recent infection rates estimated by pooling PCR were 3.27% and 3.02% respectively. The differences in recent infection rates of HIV-1 estimated by the two algorithms were not significant. Conclusions: The recent infection rates of HIV-1 estimated by LAg-Avidity EIA and pooling PCR in sentinel surveillance in MSM in Yunnan had good consistency from 2016 to 2017. Using the two methods might have a better stability in continuous surveillance for recent infection rates of HIV-1.

[Characteristics of HIV-1 genotype and drug resistance among men who have sex with men in Kunming, 2018].

Objective: To understand the characteristics of HIV-1 genotypes and drug resistance among men who have sex with men in Kunming in 2018. Methods: A total of 193 plasma samples were collected from the newly reported HIV-1 infected MSM in Kunming from January to December 2018. Viral RNA was extracted, and the gag, pol, env gene segments were amplified by nested PCR. HIV-1 genotypes and drug resistance were also analyzed. Subsequently, the evolutionary characteristics of CRF55_01B and CRF68_01B among MSM in Kunming were analyzed by Bayesian Markov Chain Monte Carlo method. Results: Multiple HIV-1 genotypes were identified among these 193 samples, including CRF07_BC (39.4%, 76/193), CRF01_AE (34.2%, 66/193), unique recombinant forms (URFs) (20.2%, 39/193), CRF08_BC (3.1%, 6/193), CRF55_01B (1.6%, 3/193), subtype B (1.0%, 2/193) and CRF68_01B (0.5%, 1/193). Results from the Bayesian evolutionary analysis showed that CRF55_01B started to spread locally after being imported from other provinces, while CRF68_01B was likely to have been brought in from the eastern provinces of China. Prevalence of HIV-1 drug resistant strains was 2.6%(5/190) before antiviral treatment, with mutation rates resistant to non-nucleoside reverse transcriptase inhibitors being the highest (2.1%, 4/190) among MSM in Kunming, 2018. Conclusion: The diversity of HIV-1 was increasing among MSM in Kunming. Although the resistance rate on pretreatment drug was relatively low, the emergence of multiple resistant strains to first-line antiviral drugs posed a challenge to antiretroviral therapy, in Kunming.

LncRNA LUCAT1 promotes proliferation of ovarian cancer cells by regulating miR-199a-5p expression.

OBJECTIVE: To investigate the biological effect of long non-coding ribonucleic acid (lncRNA) lung cancer-associated transcript 1 (LUCAT1) in the development of ovarian cancer. MATERIALS AND METHODS: Real-time quantitative polymerase chain reaction (RT-qPCR) was utilized to detect the expression levels of lncRNA LUCAT1 in three human ovarian cancer cell lines (CaoV-3, SK-OV-3 and HO-8910) and the normal human ovarian surface epithelial cell line (IOSE80). Small interfering RNAs against lncRNA LUCAT1 (si-LUCAT1) were transfected into SK-OV-3 cells. Transfection efficiency of si-LUCAT1 was verified via RT-qPCR. Cell Counting Kit-8 (CCK-8) and colony formation assays were performed to test the effect of silencing lncRNA LUCAT1 on SK-OV-3 cell proliferation. The apoptosis was measured by flow cytometry. The miRcode database was searched to predict potential microRNAs (miRNAs) binding lncRNA LUCAT1. It was found that lncRNA LUCAT1 contained a highly conserved binding site of miR-199a-5p in the 3'-untranslated region (3'-UTR). Subsequently, the targeting relationship between them was determined through Dual-Luciferase reporter gene assay and RT-qPCR analysis. RESULTS: LncRNA LUCAT1 was highly expressed in three human ovarian cancer cell lines compared to that in normal ovarian surface epithelial cell line (p<0.05). The cell proliferation rate in SK-OV-3 cells with lncRNA LUCAT1 knockdown was remarkably lower in comparison to that in control group. Moreover, colony formation assay also revealed that the number of cell clones decreased significantly after knockdown of lncRNA LUCAT1 compared to that in control group (p<0.05). In addition, the apoptosis rate was distinctly elevated in the lncRNA LUCAT1 silencing group (p<0.05). Furthermore, a highly conserved binding site of miR-199a-5p was found in the 3'-UTR of lncRNA LUCAT1. Dual-Luciferase reporter gene assay exhibited that the Luciferase activity of LUCAT1-wt was significantly reduced after overexpression of miR-199a-5p (p<0.05), while that of LUCAT1-mut was unchangeable. Further analysis via RT-qPCR suggested that miR-199a-5p overexpression significantly decreased the expression level of lncRNA LUCAT1 (p<0.05). CONCLUSIONS: LncRNA LUCAT1 is overexpressed in ovarian cancer cells, which may target miR-199a-5p to exert its effects on driving the malignant development of ovarian cancer.

MicroRNA-579-3p promotes the progression of osteoporosis by inhibiting osteogenic differentiation of mesenchymal stem cells through regulating Sirt1.

OBJECTIVE: To explore whether microRNA-579-3P was involved in the development of osteoporosis, and to investigate the possible molecular mechanisms. PATIENTS AND METHODS: The messenger RNA (mRNA) expression levels of microRNA-579-3P, alkaline phosphatase (ALP), runt-related transcription factor 2 (RUNX2) and bone sialoprotein (BSP) in serum samples of osteoporosis patients and normal controls were detected by quantitative Real-time polymerase chain reaction (qRT-PCR), respectively. Meanwhile, the expressions of the above genes during osteogenic differentiation of human bone marrow mesenchymal stem cells (hMSCs) were examined as well. To investigate the effect of microRNA-579-3P on osteogenesis, microRNA-579-3P was overexpressed and knocked down in hMSCs. Subsequently, the mRNA and protein expression levels of osteogenesis-related genes, such as ALP, RUNX2 and BSP, were detected by qRT-PCR and Western blot, respectively. In addition, ALP activity and mineralization forming ability were evaluated by ALP staining and alizarin red staining. Bioinformatics predicted that Sirt1 was the target gene of microRNA-579-3P. Subsequent luciferase reporter gene assay was performed to verify the binding relationship of microRNA-579-3P to Sirt1. Meanwhile, qRT-PCR and Western blot were used to detect the changes in the mRNA and protein expression levels of Sirt1, respectively. After overexpression of microRNA-579-3P and Sirt1, qRT-PCR, Western blot, ALP staining and alizarin red staining assays were performed to detect the osteogenic differentiation of hMSCs. RESULTS: The expression of microRNA-579-3P in serum of patients with osteoporosis was significantly higher than that of normal controls. Meanwhile, the expression of microRNA-579-3P decreased gradually during osteogenic differentiation of hMSCs. Overexpression of microRNA-579-3P significantly reduced the expressions of osteogenic related genes, including ALP, RUNX2 and BSP. Besides, ALP activity and mineralized nodule formation ability decreased obviously as well. Luciferase reporter gene assay showed that microRNA-579-3P could bind to Sirt1. After overexpression of microRNA-579-3P, the mRNA and protein expression levels of Sirt1 were significantly reduced, which were reversed after silence of microRNA-579-3P. Simultaneous overexpression of microRNA-579-3P and Sirt1 could reverse the inhibition of osteogenic differentiation of hMSCs caused by overexpression of microRNA-579-3P alone. CONCLUSIONS: MicroRNA-579-3P could inhibit osteogenic differentiation of hMSCs by regulating Sirt1, thereby promoting the development of osteoporosis.

Varroa destructor changes its cuticular hydrocarbons to mimic new hosts.

Varroa destructor (Vd) is a honeybee ectoparasite. Its original host is the Asian honeybee, Apis cerana, but it has also become a severe, global threat to the European honeybee, Apis mellifera. Previous studies have shown that Varroa can mimic a host's cuticular hydrocarbons (HC), enabling the parasite to escape the hygienic behaviour of the host honeybees. By transferring mites between the two honeybee species, we further demonstrate that Vd is able to mimic the cuticular HC of a novel host species when artificially transferred to this new host. Mites originally from A. cerana are more efficient than mites from A. mellifera in mimicking HC of both A. cerana and A. mellifera. This remarkable adaptability may explain their relatively recent host-shift from A. cerana to A. mellifera.

TLX activates MMP-2, promotes self-renewal of tumor spheres in neuroblastoma and correlates with poor patient survival.

Nuclear orphan receptor TLX (Drosophila tailless homolog) is essential for the maintenance of neural stem/progenitor cell self-renewal, but its role in neuroblastoma (NB) is not well understood. Here, we show that TLX is essential for the formation of tumor spheres in three different NB cell lines, when grown in neural stem cell media. We demonstrate that the knock down of TLX in IMR-32 cells diminishes its tumor sphere-forming capacity. In tumor spheres, TLX is coexpressed with the neural progenitor markers Nestin, CD133 and Oct-4. In addition, TLX is coexpressed with the migratory neural progenitor markers CD15 and matrix metalloproteinase-2 (MMP-2) in xenografts of primary NB cells from patients. Subsequently, we show the effect of TLX on the proliferative, invasive and migratory properties of IMR-32 cells. We attribute this to the recruitment of TLX to both MMP-2 and Oct-4 gene promoters, which resulted in the respective gene activation. In support of our findings, we found that TLX expression was high in NB patient tissues when compared with normal peripheral nervous system tissues. Further, the Kaplan-Meier estimator indicated a negative correlation between TLX expression and survival in 88 NB patients. Therefore, our results point at TLX being a crucial player in progression of NB, by promoting self-renewal of NB tumor-initiating cells and altering their migratory and invasive properties.

Optimal combination of radiotherapy and endocrine drugs in breast cancer treatment.

PURPOSE: Breast cancer is the most common cancer in women. Radiotherapy is considered a standard treatment option after surgery and adjuvant endocrine therapy is also universally used. Tamoxifen and letrozole are the current first-line endocrine therapy drugs. However, information has been scarce about how best to sequence these therapies to maximize their effectiveness and keep toxic effects to a minimum. In this study, we observed the effect of different sequence combination of radiotherapy and endocrine drugs, tamoxifen or letrozole, to get the best treatment sequence. MATERIALS AND METHODS: The combination effect of radiotherapy and tamoxifen was observed on breast tumour cell line MCF-7, radiotherapy and letrozole on aromatase-expressing breast tumour cell line MCF-7CA. Irradiation was performed with 6Gy, except for doses ranging from 0 to 8Gy for clone formation assay. Tamoxifen or letrozole was added before or after irradiation, respectively. Radiosensitivity was evaluated by clonogenic assay, cell viability by 3-(4,-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt (MTS) assay. To explore the potential mechanism, cell apoptosis was determined by DNA-binding dye 4',6-diamidino-2-phenylindole dihydrochloride (DAPI) assay, the change of Bcl-2 and Bax expression was by western blot. RESULTS: Although no significant statistical difference was observed between different sequence, tamoxifen and letrozole both increased radiosensitivity. Furthermore, the above inhibitory effect was related with apoptosis signaling pathway, especially Bcl-2 and Bax. CONCLUSION: Taken together, these results suggested that endocrine drugs, such as tamoxifen and letrozole, have potential application with radiotherapy.

Retrovirus-mediated tk gene therapy of implanted human breast cancer in nude mice under the regulation of Tet-On.

Tight regulation of the therapeutic gene expression is critical in gene therapy. In this report, a doxycycline (Dox)-regulated retrovirus-mediated gene expression system was used to study the effects of suicide gene therapy on human breast cancer cell line MCF-7 and the nude mice model of implanted human breast cancer. To render the expression of suicide gene under control, we used two pseudoviruses simultaneously, RevTRE/HSVtk and RevTet-On, to infect MCF-7 cells or xenografts of nude mice. When infected by the pseudoviruses and followed by Dox and Ganciclovir (GCV) treatment, MCF-7 cells were arrested at S phase and the growth was suppressed. We then evaluated the antitumor efficiency of this system in vivo through studying the mice bearing human breast cancer xenografts. Compared with control groups, the HSVtk mRNA level increased significantly in tumor tissues, mass of the tumors shrank remarkably, and tumor necrosis features occurred after treatment with Dox and GCV. These data suggest that suicide gene therapy using the Dox-induced Tet-On-controlled HSVtk gene expression system is a feasible method to treat human breast cancer.