BMSC-derived Exosomes Ameliorate Peritoneal Dialysis-associated Peritoneal Fibrosis via the Mir-27a-3p/TP53 Pathway.

OBJECTIVE: Peritoneal fibrosis (PF) is the main cause of declining efficiency and ultrafiltration failure of the peritoneum, which restricts the long-term application of peritoneal dialysis (PD). This study aimed to investigate the therapeutic effects and mechanisms of bone marrow mesenchymal stem cells-derived exosomes (BMSC-Exos) on PF in response to PD. METHODS: Small RNA sequencing analysis of BMSC-Exos was performed by second-generation sequencing. C57BL/6J mice were infused with 4.25% glucose-based peritoneal dialysis fluid (PDF) for 6 consecutive weeks to establish a PF model. A total of 36 mice were randomly divided into 6 groups: control group, 1.5% PDF group, 2.5% PDF group, 4.25% PDF group, BMSC-Exos treatment group, and BMSC-Exos+TP53 treatment group. Reverse transcription quantitative polymerase chain reaction (RT-qPCR) was performed to measure the expression level of miR-27a-3p in BMSC-Exos and peritoneum of mice treated with different concentrations of PDF. HE and Masson staining were performed to evaluate the extent of PF. The therapeutic potential of BMSC-Exos for PF was examined through pathological examination, RT-qPCR, Western blotting, and peritoneal function analyses. Epithelial-mesenchymal transition (EMT) of HMrSV5 was induced with 4.25% PDF. Cells were divided into control group, 4.25% PDF group, BMSC-Exos treatment group, and BMSC-Exos+TP53 treatment group. Cell Counting Kit-8 assay was used to measure cell viability, and transwell migration assay was used to verify the capacity of BMSC-Exos to inhibit EMT in HMrSV5 cells. RESULTS: Small RNA sequencing analysis showed that miR-27a-3p was highly expressed in BMSC-derived exosomes compared to BMSCs. The RT-qPCR results showed that the expression of miR-27a-3p was upregulated in BMSC-Exos, but decreased in PD mice. We found that PF was glucose concentration-dependently enhanced in the peritoneum of the PD mice. Compared with the control mice, the PD mice showed high solute transport and decreased ultrafiltration volume as well as an obvious fibroproliferative response, with markedly increased peritoneal thickness and higher expression of α-SMA, collagen-I, fibronectin, and ECM1. The mice with PD showed decreased miR-27a-3p. Peritoneal structural and functional damage was significantly attenuated after BMSC-Exos treatment, while PF and mesothelial damage were significantly ameliorated. Additionally, markers of fibrosis (α-SMA, collagen-I, fibronectin, ECM1) and profibrotic cytokines (TGF-ß1, PDGF) were downregulated at the mRNA and protein levels after BMSC-Exos treatment. In HMrSV5 cells, BMSC-Exos reversed the decrease in cell viability and the increase in cell migratory capacity caused by high-glucose PDF. Western blotting and RT-qPCR analysis revealed that BMSC-Exos treatment resulted in increased expression of E-cadherin (epithelial marker) and decreased expression of α-SMA, Snail, and vimentin (mesenchymal markers) compared to those of the 4.25% PDF-treated cells. Importantly, a dual-luciferase reporter assay showed that TP53 was a target gene of miR-27a-3p. TP53 overexpression significantly reversed the decreases in PF and EMT progression induced by BMSC-Exos. CONCLUSION: The present results demonstrate that BMSC-Exos showed an obvious protective effect on PD-related PF and suggest that BMSC-derived exosomal miR-27a-3p may exert its inhibitory effect on PF and EMT progression by targeting TP53.

Clinical characteristics and short-term mortality of 102 hospitalized hemodialysis patients infected with SARS-CoV-2 omicron BA.2.2.1 variant in Shanghai, China.

Background: The aim of this study was to analyze clinical features and short-term mortality in hemodialysis (HD) patients infected with severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) omicron BA.2.2.1 variant. Methods: In a retrospective single-center case series, 102 consecutive hospitalized HD patients infected with the coronavirus omicron variant were assessed at Pudong Hospital in Shanghai, China, from April 6 to April 18, 2022; the final date of follow-up was May 16, 2022. Clinical, laboratory, chest CT, and treatment data were collected and analyzed. The association between these factors and all-cause mortality was studied using univariate and multivariate analyses. The relationship between lymphocyte count and short-term mortality was based on receiver operating characteristic (ROC) curve analysis. Kaplan-Meier analysis was used to assess overall survival. Results: In total, 102 patients were included in this study. The patients were divided into two groups: HD patients with pneumonia (N = 46) and without pneumonia (N = 56). Of the 102 patients, 12 (11.8%) died. Multivariate regression analysis revealed that all-cause mortality was correlated with lymphocyte counts and type B natriuretic peptide (BNP), C-reactive protein (CRP), and D-dimer levels (P < 0.05). The cut-off value of lymphocyte counts was 0.61 × 109/L for all-cause mortality. The overall survival rate was significantly different between HD patients with and without pneumonia (P < 0.05). Conclusions: Lymphocyte counts are important for the prediction of short-term mortality in HD patients with SARS-CoV-2 infection. HD patients with lung involvement have poorer survival rates than those without lung involvement.

Curcumin ameliorates peritoneal fibrosis via inhibition of transforming growth factor-activated kinase 1 (TAK1) pathway in a rat model of peritoneal dialysis.

BACKGROUND: Peritoneal fibrosis (PF) remains a serious complication of long-term peritoneal dialysis (PD). The goal of this study was to investigate the anti-fibrotic effects of curcumin on the PF response to PD and its' mechanism. METHODS: Male Sprague-Dawley rats were infused with 20 mL of 4.25% glucose-based standard PD fluid for 8 consecutive weeks to establish PF model and then divided into five groups: Control, received sham operation and 0.9% physiological saline; PD, received 4.25% standard PD fluid; Curcumin, PD rats injected intraperitoeally with curcumin for 8 weeks at doses of 10, 20 or 40 mg/kg. Masson's staining was performed to evaluate the extent of PF. Peritoneal Equilibration Test (PET) was conducted to assess ultrafiltration volume (UFV) and mass transfer of glucose (MTG), quantitative RT-PCR, and immunohistochemistry or western blotting were performed to measure the expression levels of inflammation and fibrosis-associated factors. We also detected the TGF-ß1 in peritoneal fluid by ELISA. RESULTS: Compared with the control group, the PD rats showed decreased UFV (2.54 ± 0.48 to 9.87 ± 0.78 mL, p < 0.05] and increased MTG (18.99 ± 0.86 to 10.85 ± 0.65 mmol/kg, p < 0.05) as well as obvious fibroproliferative response, with markedly increased peritoneal thickness (178.33 ± 4.42 to 25.26 ± 0.32um, p < 0.05) and higher expression of a-SMA, collagen I and TGF-ß1. Treatment with curcumin significantly increased UFV, reduced MTG and peritoneal thickness of PD rats. The elevated TGF-ß1 in peritoneal fluid of PD rats was significantly decreased by curcumin. It attenuated the increase in protein and mRNA of TGF-ß1, α-SMA and collagen I in peritoneum of PD rats. The mRNA expressions of TAK1, JNK and p38, as well as the protein expressions of p-TAK1, p-JNK and p-p38 in peritoneum of PD rats were reduced by curcumin. CONCLUSIONS: Present results demonstrate that curcumin showed a protective effect on PD-related PF and suggest an implication of TAK1, p38 and JNK pathway in mediating the benefical effects of curcumin.

[Rev-erbß Knock-out Affectsthe Proliferation, Migration and Invasion Ability of Hepatocellular Carcinoma HepG2 Cell Line in vitro].

OBJECTIVE: To investigate the effect of nuclear receptor Rev-erbß knockout on proliferation and migration ability of human hepatocellular carcinoma cell line HepG2. METHODS: -The Rev-erbß gene knockout HepG2 cell line was abtained by CRISPR/Cas9 genome editing technique with specific DNA modification of the target gene. The Rev-erbß gene targeting vectors were co-transfected into HepG2 cells. Through cloning and screening, the Rev-erbß gene knockout HepG2 cell line was constructed, PCR, sequencing and Western blot methods were carried out for the identification of the Rev-erbß gene knockout HepG2 cell line. The expression level of tumor migration and invasion-associated gene in Rev-erbß gene knockout cell was determined by real-time quantitative PCR (qRT-PCR) and was compared with normal cell as control.MTT, cell scratch and Transwell experiments were conducted in order to explore the effect of Rev-erbß gene on HepG2 cell's ability of proliferation, migration and invasion. RESULTS: A Rev-erbß gene knockout monoclonal cell line, which was identified by PCR, sequencing and Western blot, was successfully constructed and named HepG2 C5 (Rev-erbß -/-). qRT-PCR results showed that Rev-erbß knockout resulted in up-regulation of matrix metalloproteinase-2 (MMP2), matrix metalloproteinase-9 (MMP9) and extracellular matrix protein-1 (ECM1) gene expression (P < 0.05) and down-regulation of E-cadherin (CDH1) gene expression (P=0.05).Results of MTT, cell scratch and transwell experiments showed that HepG2 C5 had stronger proliferation, migration and invasion ability than control cells (P < 0.05). CONCLUSION: Rev-erbß gene knockout could change the expression of migration and adhesion-associated genes in HepG2 cell, and then affect the proliferation, migration and invasion ability of HepG2 cells.

[Epidemiological features and changing patterns of intestinal parasitic diseases in Jiangxi Province].

OBJECTIVE: To understand the epidemiological features and changing patterns of intestinal parasitic diseases in Jiangxi Province from 1989 to 2014, so as to provide scientific evidence for formulating the preventive and control strategy for intestinal parasitic diseases. METHODS: The data regarding the status of intestinal parasitic diseases in Jiangxi Province were captured from the 1989 national survey on the distribution of human parasites in China and the 2014 national survey on major human parasitic diseases in China, and the status of human intestinal parasite infections was compared between the two surveys in Jiangxi Province. RESULTS: The prevalence rate of human intestinal parasite infections was 79.59% and 9.64% in Jiangxi Province in 1989 and 2014, with a decline rate of 87.89% during the 25-year period (χ2 = 30 870.5, P < 0.01). There were 24 and 20 species of human intestinal parasites detected in Jiangxi Province in 1989 and 2014, respectively, with totally 26 species detected during the two surveys. In Jiangxi Province, the 3 most highly prevalent human intestinal parasites included Ascaris lumbricoides (71.06%), hookworm (17.61%) and Enterobius vermicularis (17.59%) in 1989, and E. vermicularis (13.73%), hookworm (4.66%), whipworm (0.95%) in 2014. A higher rate of human intestinal parasite infections was found in females than in males in both surveys in Jiangxi Province (P < 0.01), and the prevalence of human intestinal parasite infections reduced by 82.50% to 95.31% in different age groups in Jiangsu Province during the 25-year period. CONCLUSIONS: The prevalence of human intestinal parasite infections shows a remarkable decline tendency in Jiangxi Province, and soil-transmitted nematodes remain the main species of human intestinal parasites in Jiangxi Province.

Curcumin suppresses epithelial-to-mesenchymal transition of peritoneal mesothelial cells (HMrSV5) through regulation of transforming growth factor-activated kinase 1 (TAK1).

OBJECTIVE: Peritoneal fibrosis remains a serious complication of long-term peritoneal dialysis (PD) leading to peritoneal membrane ultrafiltration failure. Epithelial-mesenchymal transition (EMT) of peritoneal mesothelial cells (PMCs) is a key process of peritoneal fibrosis. Curcumin has been previously shown to inhibit EMT of renal tubular epithelial cells and prevent renal fibrosis. There are only limited reports on inhibition of PMCs-EMT by curcumin. This study aimed to investigate the effect of curcumin on the regulation of EMT and related pathway in PMCs treated with glucose-based PD. METHODS: EMT of human peritoneal mesothelial cells (HMrSV5) was induced with glucose-based peritoneal dialysis solutions (PDS). Cells were divided into a control group, PDS group, and PDS group receiving varied concentrations of curcumin. Cell Counting Kit-8 (CCK-8) assay was used to measure cell viability, and a transwell migration assay was used to verify the capacity of curcumin to inhibit EMT in HMrSV5 cells. Real-time quantitative PCR and western blot were used to detect the expression of genes and proteins associated with the EMT. RESULTS: High glucose PDS decreased cell viability and increased migratory capacity. Curcumin reversed growth inhibition and migration capability of human peritoneal mesothelial cells (HPMCs). In HMrSV5 cells, high glucose PDS also decreased expression of epithelial markers, and increased expression of mesenchymal markers, a characteristic of EMT. Real-time RT-PCR and western blot revealed that, compared to the 4.25% Dianeal treated cells, curcumin treatment resulted in increased expression of E-cadherin (epithelial marker), and decreased expression of α-SMA (mesenchymal markers) (P < 0.05). Furthermore, curcumin reduced mRNA expression of two extracellular matrix protein, collagen I and fibronectin. Curcumin also reduced TGF-ß1 mRNA and supernatant TGF-ß1 protein content in the PDS-treated HMrSV5 cells (P < 0.05). Furthermore, it significantly reduced protein expression of p-TAK1, p-JNK and p-p38 in PDS-treated HMrSV5 cells. CONCLUSIONS: Our results demonstrate that curcumin showed an obvious protective effect on PDS-induced EMT of HMrSV5 cells and suggest implication of the TAK1, p38 and JNK pathway in mediating the effects of curcumin in EMT of MCs.

[Study on distribution status of Oncomelania hupensis, the intermediate host of Schistosoma japonicum in Jiangxi Province â ¡ Spatial-temporal distribution of snail-infested environments].

OBJECTIVE: To grasp the status of Oncomelania hupensis snail-infested environments in schistosomiasis endemic areas of Jiangxi Province, and establish the spatial database of snail-infested environments with the administrative village as the unit. METHODS: The spatial and temporal distributions of the historical snail environments, existing snail environments, and suspicious snail-infested environments were analyzed by the historical data review and field investigations. RESULTS: The distribution of snail-infested environments in Jiangxi Province included two popular types of lakeshore and hills. The O. hupensis snail-infested environments were mainly concentrated in the Poyang Lake area, which accounting for 66.97% of the total snail-infested areas. The potential snail-infested environment area was 204 745.48 hm2, among which the potential snail-infested environment areas in the lakeshore and hills were 146 548.58 hm2 and 58 196.90 hm2, accounting for 71.58% and 28.42%, respectively. The real existing snail area was 83 234.50 hm2, among which the real existing snail areas in the lakeshore and hills were 80 890.81 hm2 and 2 343.69 hm2, accounting for 97.18% and 2.82%, respectively. The compressed snail area was 114 253.30 hm2 with a compression rate of 57.85%, of which the compression rates in the lakeshore and hills were 46.51% and 94.97%, respectively. In the four types of snail-infested environments, the areas of Type I, Type II, Type III and Type IV accounted for 55.77%, 25.75%, 13.91% and 4.57% of the total area, respectively. CONCLUSIONS: The spatial database of O. hupensis snail-infested environments is established comprehensively and systematically, that will be conducive to tracking and performing the dynamic updates of the data of snails, so as to provide an important technical support for investigation and monitoring in the future.

[Study on distribution status of Oncomelania hupensis - intermediate host of Schistosoma japonicum in Jiangxi Province I Analysis of distribution of O. hupensis in Poyang Lake area].

OBJECTIVE: To grasp the distribution status of Oncomelania hupensis snails in Poyang Lake area, so as to provide the evidence for formulating and adjusting the schistosomiasis prevention and control strategy in lake areas. METHODS: The vector grid was created and sampled randomly by 200 m × 200 m in the spatial database of grassland, and the distribution of snails was investigated in the selected grid by using the method of mechanical sampling by 50 m × 50 m. At the same time, the elevation of investigation points was extracted based on the topographic map of Poyang Lake. RESULTS: Totally 949 and 210 investigation points were collected from the south and north of Poyang Lake areas, accounting for 3.04% and 3.21% of all the investigation points in the respective region. The number of investigation points, the appearance rate of snail frame, and the average density of alive snails were 15 231, 8.15%, and 0.463/0.1 m2, respectively. The elevation of snail distribution area of the south and north Poyang Lake areas were 11-16 m and 9-16 m respectively. The elevation of concentrated snail belts of the south Poyang Lake area were 12-13 m and 15-16 m, and the elevation of concentrated snail belts of the north Poyang Lake area was 12-14 m. CONCLUSIONS: The distribution of snails is in the range of 9-16 m. The suitable habitats of snail breeding are moving from the south Poyang Lake area to the north Poyang Lake area, and from high elevation to low elevation. In the future, the schistosomiasis prevention and control measures could be formulated based on the geographical characteristics of current snail distribution in order to consolidate the achievements of schistosomiasis control.

[Trends of soil-transmitted nematode infections in Jiangxi Province].

OBJECTIVE: To analyze the status and trends of soil-transmitted nematode infections in Jiangxi Province from 1989 to 2014, so as to provide the evidence for generating the strategy of soil-transmitted nematode prevention and control. METHODS: The data of three epidemiological surveys on human parasitic diseases (in 1989, 2002 and 2014) were classified and analyzed. The stool examination by Kato-Katz's thick smear method was adopted for the investigation of soil-transmitted nematode infections. RESULTS: The total infection rate of soil-transmitted nematodes decreased by 91.89% from 77.67% in 1989 to 6.30% in 2014, in which the infection rate of Ascaris lumbricoides decreased by 98.78% from 71.11% to 0.87%, the infection rate of Trichuris trichiura decreased by 96.80% from 29.67% to 0.95%, and the infection rate of hookworm declined by 73.57% from 17.63% to 4.66%. The infection rates of soil-transmitted nematodes in the female were higher than those in the male in three surveys. In different ecological districts, the infection rates of soil-transmitted nematodes in the female were also higher than those in the male, except in Zhe-Min Ecological District in 2002 and 2014. A declined trend of the infection was showed in all age-groups in the three surveys, but it slowed down by the growth of age, i.e., the reduction rate was 97.03% in the age group of < 10 years while 80.62% in the age group of >70 years. In 2014, the number of persons infected with soil-transmitted nematodes occupied 65.4% of the whole number of persons infected with intestinal parasites. CONCLUSIONS: The mean infection rates of soil-transmitted nematodes decrease obviously in human population in different ecological districts, but the soil-transmitted nematodes are still the main species in intestinal parasite infections. The sequence of dominant species changes from A. lumbricoides, hookworm and T. trichiura in 1989 to hookworm, T. trichiura and A. lumbricoides in 2014. The rural female and elder people are the key population, while hookworm is the key species for the prevention and control of soil-transmitted nematodes.

[Study on cut-off value of IHA method for schistosomiasis diagnosis in different endemic areas].

OBJECTIVE: To explore the cut-off value of the indirect haemagglutination test (IHA) method for schistosomiasis japonica diagnosis in different endemic areas. METHODS: Totally 55 nature villages of the lake-type endemic counties, Yugan and Xinzi, in Poyang Lake Region of Jiangxi Province were chosen as the study fields, and all the villagers over 5 years old were parallelly examined by Kato-Katz method + miracidial hatching test and IHA method. The detection data were analyzed by the correlation analysis, and the threshold values of the IHA method in different endemic areas were decided by the receiver operating characteristic (ROC) curve. RESULTS: The positive rate of stool examinations of the villagers was correlated with the distribution trend of the antibody level of whole population (r = 0.588, P ＜ 0.05), but no correlation with the antibody level of the positive population (r = 0.221, P ＞ 0.05). The antibody level of stool-negative population during the period of 2008 to 2011 detected by IHA method dropped year by year, and the annual difference was statistically significant (F = 3.650, P ＜ 0.05). While the antibody level of stool-positive population found during the period of 2008 to 2011 maintained a certain high level in the 4 years, and there was no statistically significant difference among them (F = 2.461, P ＞ 0.05). When the positive rates were <1%, 1%-5% or >5%, the specificity of diagnosis could be improved when 1∶80, 1∶20 and 1∶10 were used as the cut-off values of IHA correspondingly. CONCLUSIONS: The different threshold values for diagnosis of schistosomiasis japonica should be considered while using IHA method to screen out patients in different endemic areas.

[Analysis of infective status of Clonorchis sinensis in Jiangxi Province].

OBJECTIVE: To understand the current prevalence of clonorchiasis in Jiangxi Province. METHODS: A survey was performed according to the scheme of the 3rd Principal Human Parasites of Jiangxi Province. Based on the ecological regions, a stratified cluster sampling method was applied by the economic and geographic situation. In rural areas, the investigation of C. sinensis was carried out together with the soil-transmitted helminths investigation, and in the urban areas, the random cluster sampling method was applied for the C. sinensis investigation. There were 92 survey sites from 32 counties. The eggs of C. sinensis in stool were examined by Kato-Katz technique, and health knowledge was also investigated by questionnaires in some people at the same time. RESULTS: A total of 23 606 sample residents were investigated, and 138 were found infected with C. sinensis, with the infection rate of 0.58%. Light infection was found in most of them. Totally 124 C. sinensis infected persons focused in Xinfeng County, and only a few of infected people scattered in the other counties. In Xinfeng County, 851 residents were investigated. Among them, the infected people were found in all the age groups except the 0- year age group. The highest infection rate appeared in the 70- years group (24.00%). The male infection rate was 20.29%, which was higher than that of the female (6.25%), showing a statistically significant difference (P＜0.01). The infection rate was highest in the population who received high school or technical secondary school education (31.48%). For the occupation distribution, the infection rate was highest in public officers (39.39%). The questionnaire survey showed that the infection rate in the populations in Xinfeng County who had the history of eating raw fish or raw shrimp was 33.15%. CONCLUSIONS: The distribution of C. sinensis infection presents a regional aggregation in Xinfeng County, but in other areas, the distribution is sporadic. It is necessary to continue to carry out the parasitic disease screening, and in Xinfeng County, it is necessary to strengthen the comprehensive prevention and control intervention.

[Correlation of the second to fourth digit ratio with the body mass index of infertile men].

OBJECTIVE: To investigate the relationship between the second to the fourth digit ratio (2D:4D) and body mass index (BMI) in infertile men of the Han ethnic group in Ningxia. METHODS: Using anthropometry, we calculated the mean ratio of 2D:4D and BMI of 197 infertile men and 148 normal healthy male controls, followed by analysis of their relationship. RESULTS: The BMI was correlated positively with the 2D:4D ratio of the left hand in the infertile men (P < 0.05) and in the patients with a higher 2D:4D ratio of the left hand (P < 0.05), but negatively with the 2D:4D ratio of the righ/left (Dr-1) (left: P < 0.01; Dr-l: P < 0.05). The mean 2D: 4D ratio and BMI were both lower in the normal control than in the infertile men, with statistically significant differences in BMI (P < 0.05) and the 2D:4D ratio of the left hand (P < 0.05). CONCLUSION: There is a correlation between the 2D:4D ratio and BMI in infertile men.

Prevalence and molecular characterization of fluoroquinolone resistance in Escherichia coli isolates from dairy cattle with endometritis in China.

Fluoroquinolones are frequently used to treat infectious disease that is caused by Escherichia coli in dairy cattle. However, fluoroquinolone resistance occurs and is due either to chromosomal mutations in the bacterial topoisomerase genes and/or to plasmid-mediated resistance genes. The purpose of this study was to determine the prevalence and molecular characteristics of fluoroquinolone resistance determinants in E. coli strains (n=148) isolated from dairy cattle with bovine endometritis in Inner Mongolia (China). Analysis of the mutations in the quinolone resistance-determining regions of resistant E. coli isolates confirmed previously reported substitutions in the GyrA and ParE. However, we identified additional substitutions in the ParC and GyrB that have not been reported earlier. No plasmid-mediated quinolone resistance genes in any of the isolates were found. The number of point mutations found per isolate correlated with an increase in the minimum inhibitory concentration of ciprofloxacin. Overall, 45.5% of the isolates were positive for the class I integrase gene along with four gene cassettes that were responsible for resistance to trimethoprim (dfr1 and dfrA17) and aminoglycosides (aadA1 and aadA5), respectively. The prevalence of extended-spectrum ß-lactamases (ESBLs) was 100%, and the blaTEM gene was predominant in all of the isolates. In conclusion, our results identify the mechanism of quinolone resistance for the first time and reveal the prevalence of integron and ESBLs in E. coli isolates from dairy cattle with bovine endometritis in China after 20 years of quinolone usage in cattle.

Correlations between digit ratio and infertility in Chinese men.

BACKGROUND: Digit ratio, especially second to fourth digit ratio (2D:4D) is lower in men than in women. The ratios of digit may be established in utero and is negatively correlated with sperm counts and testosterone in men and positively correlated with estrogen in men and women. AIM: To study whether the digit ratio (especially 2D:4D) are associated with the a+b ratio of sperm number in Chinese populations. METHODS: Photocopies of the two hands of 268 men (controls: 72; patients: 196) and seminal parameters of masturbatory semen samples were collected. RESULTS: The mean values of digit ratio of the controls and the patients all presented a trend as 2D:3D<2D:4D<3D:4D<2D:5D<4D:5D<3D:5D; the patients have higher mean values than controls; significant variances of 2D:3D, 3D:4D (left: P<0.05; right: P ≤ 0.05) and 2D:4D (left: P<0.001; right: P<0.01) were found between two groups; there was a higher percentage of 2D>4D in the patients; the relationship between 2D:4D and a+b ratio of sperm number in the patients was significant (P<0.001). CONCLUSION: Digit ratio, especially 2D:4D in the left hand maybe one of the important markers of infertility in men for early diagnosis.

[Digit length ratio and male fertility among infertile patients in Ningxia].

OBJECTIVE: To investigate the association between the digit length ratio (DLR) and male fertility in Ningxia. METHODS: Using an electronic sliding caliper, we measured the lengths of the index finger (D2), middle finger (D3), ring finger (D4) and little finger (D5) of both the right and left hand of 136 infertile males in Ningxia, 45 of them with normal and the other 91 with abnormal semen, including 28 cases of oligozoospermia, 10 cases of obstructive azoospermia (OA) and 53 cases of nonobstructive azoospermia (NOA). We calculated the DLRs of D2:D3, D2:D4, D2:D5, D3:D4, D3:D5 and D4:D5 and compared them among different groups. RESULTS: The mean values of FLR presented a pattern of D2:D3 < D2:D4 < D3:D4 < D2:D5 < D4:D5 in the infertile males. DLRs of D2:D3, D2:D4, D2:D5, D3:D5 and D4:D5 of the right hand were significantly higher in the abnormal than in the normal semen group (t = 2.56, 2.48, 3.57, 2.85, 2.53; P < 0.05); those of D2:D3, D2:D4, D2:D5 and D4:D5 of the right hand presented a pattern of NOA > oligozoospermia > OA > normal semen group, while that of D3:D5 presented a pattern of NOA > OA > oligozoospermia > normal semen group. The NOA group showed a significantly higher DLR of the right hand than the normal semen group in D2:D3, D2:D4, D3:D5 and D4:D5, the oligozoospermia group in D2:D4, D3:D5 and D4:D5, and the OA group in D2:D3 (P < 0.05). The D4:D5 DLR of the right hand was remarkably higher in the Han than in the Hui ethnic group (t = 2.01, P < 0.05). CONCLUSION: Higher DLRs of D2:D3, D2:D4, D2:D5, D3:D5 and D4:D5 of the right hand were associated with lower fertility in infertile males in Ningxia. And the D4:D5 FLRs of the right hand may be different between Hui and Han infertile men in Ningxia.

Chinese human immunodeficiency virus-1 patients with different routes of transmission exhibit altered expression levels of blood dendritic cell subpopulations.

Dendritic cells (DCs) play a pivotal role in the pathogenesis of human immunodeficiency virus-1 (HIV-1). Reduced numbers of blood DCs have been observed in individuals with chronic HIV-1 infection. In the present study, we analyzed the expression levels of monocytes, myeloid dendritic cell (mDC) precursors, mDCs, and plasmacytoid dendritic cells (pDCs), in HIV-1-infected patients in China who were infected via different routes of transmission, including heterosexual and homosexual sexual contact, and blood transmission through importation of blood or blood products, to further elucidate their role in HIV. Compared with HIV-negative individuals (n = 40), relative levels of CD11c+CD14⁻mDCs, CD11c++CD123(low) mDCs, and CD11c⁻CD123+ pDCs in total peripheral blood mononuclear cells (PBMCs) were significantly lower in all HIV patients (n = 93), and in those with blood transmission (n = 26) and heterosexual transmission (n = 43), while relative levels of CD11c+CD14⁻mDCs were significantly lower in HIV patients infected via homosexual transmission (n = 24). The results of correlation analysis demonstrated a significant negative correlation between CD4+ T-cell counts and the relative levels of CD11c++CD123(low) mDCs in HIV-I patients infected via blood transmission. There was no significant correlation between CD4+ T-cell counts and the expression level of other DC subpopulations in PBMCs from HIV patients. The results of this study suggest that HIV-1 patients with different routes of transmission exhibit altered expression levels of blood DC subpopulations, which contributes to dysregulated immune responses and pathogenesis of HIV-1.

[Study on construction of chimeric adenovirus vector Ad5/11 carrying human eGFP and endostatin-K5 and its experimental investigation in vitro].

AIM: To construct chimeric adenoviral vector Ad5/11 carrying reporter gene eGFP and human endostatin-K5. METHODS: Chimeric adenoviral backbone vector expressing eGFP was generated by overlap PCR and homologous recombination in E.coli BJ5183. Then chimeric adenoviral vector Ad5/11-E1-CMV-endo-K5/E3-CMV-eGFP carrying eGFP and human endostatin-K5 was constructed by co-transfecting Pac I linearized chimeric adenoviral backbone and adenoviral E1 shuttle vector expressing human endostatin-K5 into HEK 293 cells. The expression of eGFP was observed under fluorescent microscope. The expression of human endostain-K5 in U87MG cells infected by chimeric adenoviral vector was detected by RT-PCR. The infection efficiency between chimeric adenovirus and unmodified control adenovirus for human glioblastoma cell line A172 and breast cancer cell line MDA-MB-231 in vitro was evaluated by the comparison of the expression of eGFP. RESULTS: Chimeric adenovirus Ad5/11-E1-CMV-endo-K5/E3-CMV-eGFP could successfully express eGFP and endostatin-K5. Chimeric adenoviral vector significantly enhances the infection efficiency for human glioblastoma cell line A172 and breast cancer cell line MDA-MB-231 compared with unmodified adenoviral vector Ad5 E1-CMV-eGFP. CONCLUSION: Chimeric adenoviral vector Ad5/11-E1-CMV-endo-K5/E3-CMV-eGFP can significantly improve the infection efficiency for human glioblastoma cell line A172 and breast cancer cell line MDA-MB-231.

[Characteristics of glucose metabolism in non-obese and obese women with polycystic ovarian syndrome].

OBJECTIVE: To investigate characteristics of glucose metabolism of non-obese and obese women with polycystic ovary syndrome (PCOS). METHODS: From May 2006 to April 2009, 1928 PCOS patients treated in Reproductive Medicine Center of Shandong Provincial Hospital Affiliated to Shandong University were enrolled in this study, which were divided into 901 cases [body mass index (BMI) ≥ 25 kg/m²] in obese group and 1027 cases in non-obese (BMI < 25 kg/m(2)) group. The prevalence of type 2 diabetes mellitus (T2DM), oral glucose tolerance test, impaired fasting glucose (IFG), impaired glucose tolerance (IGT) were compared between the two groups. RESULTS: (1) Blood glucose levels: at the time of fasting, 30, 60, 120 and 180 minutes, the levels of glucose were (5.3 ± 1.1), (9.0 ± 2.4), (9.3 ± 4.4), (7.5 ± 2.8), (5.3 ± 1.8) mmol/L in obese group and (5.0 ± 0.8), (8.4 ± 3.5), (8.0 ± 4.2), (6.5 ± 3.2), (4.9 ± 1.6) mmol/L in non-obese group, which all showed statistical difference at every time point (P < 0.01). (2)The level of insulin: at the time of fasting, 30, 60, 120 min, the level of insulin were (13 ± 7), (81 ± 51), (102 ± 65), (83 ± 63) mU/L in obese group and (8 ± 5), (57 ± 35), (62 ± 44), (46 ± 39) mU/L in non-obese group, which all showed statistical difference at every time point (P < 0.01). However, at time point of 180 minutes, the level of insulin did not exhibit significantly difference between obese and non-obese group (P > 0.05). (3) The prevalence of abnormal glucose metabolism: the rate of IFG was 4.98% (96/1928). The rate of abnormal glucose tolerance was 23.08% (445/1928). The rate of IGT were 13.05% (134/1027) in non-obese group and 24.20% (218/901) in obese group, which also showed remarkable difference (P < 0.01). The rate of T2DM were 2.53% (26/1027) in non-obese group and 7.44% (67/901) in obese group, which reached significant difference (P < 0.01). CONCLUSION: Abnormal glucose metabolism was observed more frequently in overweight or obese PCOS women.

[Effects of control-releasing arsenic trioxide-eluting stent on intimal smooth muscle cells and type III collagen in canine coronary artery post-stent model].

OBJECTIVE: To study the safety and efficacy of control-releasing arsenic trioxide (As2O3)-eluting stent on intimal smooth muscle cells (SMC) and type III collagen (CIII) in canine coronary artery post-stent model. METHODS: Twenty-four experimental canines were equally divided into 4 groups, the three tested groups were deployed by stents with different dosage of As2O3 (1.6 microg/mm2, 2.4 microg/mm2 and 3.2 microg/mm2 in low, median and high dose groups, respectively) and coated with polybutyl methacrylate/nano silica and poly-lactide-coglycolide in mild oversizing (stent/vessel ratio of 1.3:1) in left anterior descending (LAD) or circumflex coronary arteries (LCX), while the control group only by simple coated stent without As2O3. The effect was assessed 4 weeks after stent implantation in terms of vascular histomorphology, and changes of SMC and C III expressions were detected using immunohistochemical analysis. RESULTS: Subintimal hemorrhage, medial/adventitial necrosis, thrombosis and inflammatory cell infiltration were not found and integral endothelium could be seen under screening electron microscopy in all groups. Positive expression of SMC and CIII in the tested groups, especial in the high dose As2O3 group, was more weaker than that in control group. Histo-morphological analysis showed that the neo-genetic intimal area and vascular stenosis were lower, but the mean luminal diameter was larger in the three tested groups than that in the control group (P < 0.01). Comparisons of various indices between tested groups treated by different doses of As2O3 showed that the difference between high/median dose vs. low dose was significant (P < 0.01), but that between high dose vs. median dose was insignificant (P > 0.05). CONCLUSION: Control-releasing As2O3-eluting stent shows a reliable and safe effect in preventing and treating post-stent restenosis by its dose-dependent inhibition on expressions of SMC and CIII to suppress the neo-genesis of intimal hyperplasia.