Objective To study the effect of calcium on human peritoneal mesothelial cells (HPMCs). Methods Proliferation abilities of HPMCs were assessed by tetrazolium salt colorimetry assay (MTT assay) and the levels of LDH in the supernatant were detected in all groups to evaluate the damage of HPMCs. The expression of TNF-α in cytoplasm was detected by immunohistochemistry. Results Calcium enhanced proliferation of HPMCs in a time-dependent manor(P＜0.01), especially calcium with 1.25 mmol/L(0.5098±0.016,0.6763±0.048) and 1.0 mmol/L(0.4853±0.016,0.6678±0.076). Calcium with different concentrations significantly increased the levels of LDH in HPMCs in a time-dependent manor, while the effect of calcium with 1.25 mmol/L was lowest(17.78±1.18,23.60±1.39,P＜0.01). Calcium with 2.0 mmol/L[(42.61±3.29)%] and 1.75 mmol/L[(33.32±1.88)%] significantly up-regulated the expression of TNF-α(P＜0.01) . Conclusions High calcium damaged HPMCs and up-regulated the expression of TNF-αby HPMCs, while physical calcium (1.25 mmol/L)can protect peritoneum and prevent peritoneal fibrosis.

Objective To study the effect of PPAR-r agonist Troglitazone on the mPGEs expression induced by high glucose in rat mesangial cells. Methods Rat mesangial cells were incubated with high glucose at presence or absence of different concentrations of Troglitanzone, and the mPGEs concentration in supernatant was measured by an enzyme-linked immunosorbent assay (ELISA). Results 5% glucose could obviously induce the mPGEs expression in the rat mesangial cells (P

Objective To investigat the symptom burden of patients with maintenance hemodialysis (MHD) and explore the influencing factors of them,in order to provide the basis for improving the quality of life of these patients.Methods A total of 230 patients with MHD were enrolled in Xiangtan Central Hospital from January 2016 to December 2016,the general condition and disease-related data of them were collected,and the symptom burden of them over the past week was assessed by improved symptom burden scale.Results ① The average age of these patients were (56.8 ± 14.1)years,the sex ratio for men and women was 1.37∶ 1,the median time of dialysis was 24 months,and the frequency of dialysis was (9.16 ± 2.36)times;② The symptom score in MHD patients was (65.72 ± 28.46)points,the top 5 symptoms which had higher incidence were fatigue (72.2％),dry mouth (63.5％),itching (61.3％),falling asleep difficultly (54.3％),drying (49.2％),the top 5 symptoms which were more troubling were restless legs syndrome (2.54 ± 0.73),falling asleep difficultly (2.48 ± 0.83),bone/joint pain (2.45 ± 0.69),fatigue (2.31 ±0.77),easy to wake up (2.16 ±0.78);③ Age,sex,occupation,dialysis age,inorganicphosphorus and serum calcium had an effect on the symptom burden of MHD patients (P ＜ 0.05).Condusions We should focus on patients who are older,female,retired or no occupation,longer dialysis age,calcium or phosphorus metabolism disorders,provide targeted care and treatment,in order to reduce the symptom burden and improve the life quality of them.

Objective To detect the expression of serum microRNA (miR)-192 in diabetic nephropathy (DN) patients and investigate its effects on DN. Methods The serum levels of miR-192 and miR-210 in DN patients,diabetic patients without renal injury and healthy people were detected by real-time quantitative PCR.Down-stream target genes were predicted using TargetScan software and further confirmed by gain of function and loss of function studies in vitro.Mesangial cells were treated with different concentrations of TGF-β1,then miR-192 expressions of cells and supernatant were examined as well. Results Serum miR-192 level significantly decreased in DN patients compared with healthy people (0.41 ±0.09 vs 1.00±0.00,P＜0.01) and diabetic patients without renal injury,while the serum miR-210 levels were not significantly different among three groups.Bioinformatics analysis results showed that some target genes were involved in TGF-β signal pathway.ZBP1,IGF-1 and type Ⅰ collagen (Col Ⅰ ) were chosen and confirmed by Western blotting,which were regulated by miR-192.TGF-β1 decreased the expression of miR-192 in both cells and cell culture supematants. Conclusion Serum miR-192 may promote the progress of DN by regulating TGF-β1 signaling pathway and may be used as a potential biomarker in the diagnosis of DN depending on its certain specificity.

Objective To explore the application value of detection of Mycoplasma,Chlamydia and anti-sperm antibodies for male infertility.Methods Culture method,immune chromatography,and enzyme linked immunosorbent assay were adopted for the detection of Mycoplasma,Chlamydia,and anti-sperm antibodies respectively in 102 cases of infertile males and 42 cases fertile males.And the routine semen analysis was proceed as well.All the subjects were divided into 4 groups according to the detection re-sults:simple Mycoplasma or/and Chlamydia positive group(71 cases),simple anti-sperm antibodies positive group(21 cases),My-coplasma and Chlamydia or/and anti-sperm antibodies positive group(8 cases),Mycoplasma,Chlamydia and anti-sperm antibodies negative group(44 cases).The main indexes of semen routine were compared among 4 groups.Results The positive rates of Myco-plasma,Chlamydia and anti-sperm antibodies in infertile males were significantly higher than those of fertile males (P <0.05).The sperm densities,activity rates,activity of simple Mycoplasma or/and Chlamydia positive group,simple anti-sperm antibodies posi-tive group,and Mycoplasma and Chlamydia or /and anti-sperm antibodies positive group were significantly lower than the nega-tive group,while the sperm malformation rates,liquefaction times,white blood cell counts of the three groups were significantly higher than the negative group(P <0.05 ).Conclusion Mycoplasma,Chlamydia infection and anti-sperm antibodies production have significant effect on the indexes of semen,which cause decline in semen quality and the occurrence of male infertility.

Objective To make sure the effect of dialysate composition on HPMCs.Methods Cell strains were subculturing.There are six groups in this experiment: Group 1(control);group 2(4.25% Glucose);group 3(1.75mmol/L Ca~(2+));group 4(1.25 mmol/L Ca~(2+));group 5(4.25% Glucose+1.75mmol/L Ca~(2+));group 6(4.25% Glucose+1.25mmol/L Ca~(2+)).The capacity of proliferation of HPMCs was assessed by MTT assay. Results Proliferation of HPMCs was inhibited in 4.25% glucose group in time dependence.Ca~(2+) induce proliferation of HPMCs,however,no effect on proliferation of HPMCs exists in different Ca~(2+) group.Conclusion High glucose can inhibit cell proliferation;Ca~(2+)(1.75mmol/L,1.25mmol/L) can promote the proliferation and 1.25mmol/L is better.

Objective: To observe an abnormal expression of humoral immune response induced by memory B cells in tonsils and peripheral blood of patients with IgA nephropathy ( IgAN) , the variation of memory B cells after tonsillectomy , and to discover the role of tonsillectomy in IgAN .Methods: In the study , 28 patients were diagnosed as IgAN via renal biopsy , and 27 patients suffering from chronic ton-sillitis without nephritis and 10 normal human beings were selected as controls .The expression of memory B cells in the tonsils and peripheral blood was tested by flow cytometry , and the same method was used to test the variation of the expression of memory B cells in peripheral blood of patients with IgAN after tonsil -lectomy.Results:In this study , higher percentages of memory B cells were observed in tonsil and pe-ripheral blood of IgAN patients, which were 5.72%±5.26%, 4.92%±5.10%.After tonsillectomy, the percentage of memory B cells was 1 .10%±0 .65%, lower than that before tonsillectomy ( P <0.05).Meanwhile, in tonsils and peripheral blood , the percentage of memory B cells varied with the variation of the urinary findings of the IgAN patients .Conclusion:The percentage of memory B cell in tonsils and peripheral blood could predict disease progression of IgAN to a certain extent .

Object To investigate the chemical constituents of Epimedium koreanum Nakai.Methods Separation was performed by HPLC on a Merck LiChroCAR analytical column with the mobile phase consisting of acetonitrile-water-acetic acid as gradient eluent at the flow rate of 1 mL/min, the UV detection was set at 270 nm and TIC was recorded by an electrospray ionization mass spectrometer in positive mode. Results Nine compounds were identified by HPLC-MS2. Conclusion The method is rapid and accurate to identify the chemical constituents of the natural product.

Objective To investigate the effects of connective tissue growth factor (CTGF) siRNA delivered by pRetro-Super (PRS) retrovirus vector on extracellular matrix and VEGF expression in human peritoneal mesothelial cells (HPMC). Methods Four pairs of oligonucleotides including 64 bp DNA were designed and synthesized in vitro according to siRNA target sequence and PRS retrovirus desire.PRS-CTGF-siRNA1-4 recombinant retrovirus vectors were constructed.The recombinant retrovirus vectors containing CTGF-siRNA were transferred into PT67 packaging cell lines with lipefectamine 2000,then infected HPMC.mRNA expression was determined by semi-quantitative RT-PCR and protein expression was determined by Western blot.Results Both mRNA and protein expressions of CTGF,FN,Col I,laminin (LN) and VEGF were significantly increased in HPMC with 5 μg/L TGF-β1 stimulation (P＜0.01,respectively).CTGF,FN,Col I,LN mRNA and protein and VEGF mRNA expression stimulated by TGF-β1 were significantly decreased in HPMC infected with PRS-CTGF-siRNA1～4 retrovirus vectors (P＜0.01,respectively).The inhibitory rates on CTGF were 69.3%,22.2%,27.4% and 38.8%,respectively (P＜0.01).At the same time,there was also a significant reduction of VEGF protein expression in HPMC infected with PRS-CTGF-siRNA1 vector (P＜0.01).There was no significant difference in HPMC infected with PRS void vector. Conclusion CTGF siRNA delivered by PRS retrovirus vector can effectively inhibit the enhancement of extracellular matrix and VEGF expression stimulated by TGF-β1 in HPMC.

Objective To investigate the efficiency and safety of ultrasound-mediated microbubble destruction in enhancing β-galactosidase gene (β-gal gene) transfer into human proximal tubular cells(HKCs). Methods β-gal gene was transfected to HKCs as a mark gene with ultrasound-mediated microbubble destruction. Cultured HKCs were grouped to receive the following 7 treatments respectively: ultrasound alone; microbubble alone; naked plasmid; ultrasound and plasmid; microbubble and plasmid; ultrasound, microbubble, and plasmid; and VigoFect and plasmid. In Group 6, HKCs were exposed to ultrasound under different sound intensities and time. X-gal stainning, typan blue stainning, and Hochest stainning were used to detect the transfection efficiency, cell survival rate, and cell apoptosis rate, respectively.Results β-galactosidase expression could be observed in the ultrasound-mediated microbubble destruction groups. Along with the increasing of sound intensity and exposure time, the cell survival rate of HKCs decreased, and the cell apoptosis rate increased gradually. The transduction efficiency and survival rate in middle intensity (0.3 W/cm~2×60 s) of ultrasound exposure were higher than those of other groups, similar to those of Group 7.Conclusion Under optimum sound intensity and exposure time, ultrasound-mediated microbubble destruction can increase gene transfer into HKCs. This non-invasive gene transfer method may be a useful tool for clinical gene therapy.