Long non-coding RNA C2dat1 involved in diabetic renal interstitial fibrosis by influencing CaMK2D/NF-κB signaling pathway / 中华微生物学和免疫学杂志

Objective:To study the changes in long non-coding RNA C2dat1 expression in kidney tissues of rats at different stages of diabetic kidney disease (DKD) and its relationship with renal interstitial fibrosis.Methods:Forty-eight male SD rats were randomly divided into two groups with 24 rats in each group: control group and DKD group. The rats in the control group were fed with ordinary diet, while those in the DKD group were fed with high-fat diet and drank water freely. After eight weeks of feeding, the rats were fasted for 12 h with free access to water. Then, the DKD group was given a one-time intrabitoneal injection of streptozotocin and the control group was given an equal dose of sodium citrate buffer. After 72 h, the random peripheral blood glucose concentration (≥ 16.7 mmol/L for three consecutive days) and urine sugar (positive) were tested to assess the establishment of the diabetes model. Urine, blood and kidney samples were collected at 3, 6, 9 and 12 weeks. The urinary protein excretion rate within 24 h, urinary creatinine and serum total cholesterol were measured by automatic biochemical apparatus. Pathological changes in kidney tissues were observed by HE staining. The expression of calcium/calmodulin-dependent protein kinase Ⅱ delta (CaMK2D), p65, p50, α-SMA and E-cardherin was detected by immunohistochemistry. Quantitative real-time PCR (qPCR) was used to detect the expression of lncRNA C2dat1 and CaMK2D. The relationship of lncRNA C2dat1 with α-SMA, E-cardherin and CaMK2D was analyzed by correlation analysis. In in vitro experiment, renal tubular epithelial cells HK-2 were induced by high glucose. The expression of lncRNA C2dat1 and CaMK2D in HK-2 cells was detected by qPCR after 24, 48 and 72 h of intervention. Results:The rats in the DKD group showed typical symptoms such as polydipsia, polyphagia, significant weight loss and increased blood glucose as compared with the rats in the control group. Results of the biochemical tests revealed that compared with the control group, the DKD group had increased 24 h excretion rate of urinary protein, decreased urinary creatinine and up-regulated total cholesterol. HE staining showed that the rats in the control group had intact glomeruli, normal basement membrane and no mesangial hyperplasia or inflammatory cell infiltration. However, enlarged glomeruli and evenly thickened basement membrane were observed in the DKD group. Immunohistochemistry indicated that the expression of CaMK2D, p50 and α-SMA was higher in the DKD group than in the control group, while the expression of E-cardherin was lower in the DKD group. qPCR results showed that the expression of lncRNA C2dat1 and CaMK2D at mRNA level was higher in the DKD group than in the control group. In in vitro experiment, the expression of lncRNA C2dat1 and CaMK2D at mRNA level was also higher in HK-2 cells induced by high glucose than in the control group. Correlation analysis indicated that lncRNA C2dat1 was positively correlated with α-SMA and CaMK2D, but negatively correlated with E-cardherin. Conclusions:During the progression of DKD, the high expression of lncRNA C2dat1 might promote diabetic renal interstitial fibrosis by regulating the expression of CaMK2D to activate the NF-κB signaling pathway.

Association of lean tissue index with arteriovenous fistula dysfunction in maintenance hemodialysis patients / 中华肾脏病杂志

The clinical data of maintenance hemodialysis (MHD) patients from twenty hemodialysis centers in Guizhou province from June to September 2020 were collected by cross-sectional study. The patients were divided into AFD group and non-AFD group according to whether AFD had occurred. LTI was measured by body composition monitor. The results showed that the incidence of AFD in 2 781 MHD patients was 30.0% (835/2 781). Median LTI level was 15.2 (13.2, 17.5) kg/m 2. The LTI level in the AFD group was higher than that in the non-AFD group ( P < 0.05). According to the tertiles of LTI, low LTI group (LTI ≤ 13.9 kg/m 2) had the highest incidence of AFD (35.5%, 334/940), and the high LTI group had the lowest incidence of AFD (26.3%, 241/916), and the difference among the three groups was statistically significant ( χ2=20.182, P < 0.001). Multivariate logistic regression analysis showed that low LTI group as the reference, the risk of AFD in moderate LTI group (13.9 kg/m 2 < LTI ≤ 16.6 kg/m 2) and high LTI group were associated with the 20.0% ( OR=0.800, 95% CI 0.650-0.986, P=0.036) and 22.8% ( OR=0.772, 95% CI 0.616-0.966, P=0.024) decrease, respectively. These results suggest that low LTI level is independently associated with an increased risk of AFD in MHD patients.

1, 25-(OH)2-VitD3 attenuates renal tubulointerstitial fibrosis in diabetic kidney disease by inhibiting Snail1-SMAD3/SMAD4 complex formation / 细胞与分子免疫学杂志

Objective To investigate the effect of 1, 25-(OH)2-VitD3 (VitD3) on renal tubuleinterstitial fibrosis in diabetic kidney disease. Methods NRK-52E renal tubular epithelial cells were divided into control group (5.5 mmol/L glucose medium treatment), high glucose group (25 mmol/L glucose medium treatment) and high glucose with added VitD3 group (25 mmol/L glucose medium combined with 10-8 mmol/L VitD3). The mRNA and protein expression of Snail1, SMAD3, SMAD4, α-SMA and E-cadherin in NRK-52E cells were detected by real-time quantitative PCR and Western blot analysis respectively. The expression and localization of Snail1, SMAD3 and SMAD4 were detected by immunofluorescence cytochemical staining. The binding of Snail1 with SMAD3/SMAD4 complex to the promoter of Coxsackie-adenovirus receptor (CAR) was detected by chromatin immunoprecipitation. The interaction among Snail1, SMAD3/SMAD4 and E-cadherin were detected by luciferase assay. Small interfering RNA (siRNA) was used to inhibit the expression of Snail1 and SMAD4, and the expression of mRNA of E-cadherin was detected by real-time quantitative PCR. SD rats were randomly divided into control group, DKD group and VitD3-treated group. DKD model was established by injection of streptozotocin (STZ) in DKD group and VitD3-treated group. After DKD modeling, VitD3-treated group was given VitD3 (60 ng/kg) intragastric administration. Control group and DKD group were given normal saline intragastric administration. In the DKD group and VitD3-treated group, insulin (1-2 U/kg) was injected subcutaneously to control blood glucose for 8 weeks. The mRNA and protein levels of Snail1, SMAD3, SMAD4, α-SMA and E-cadherin in renal tissues were detected by real-time quantitative PCR and Western blot analysis respectively. Immunohistochemistry was used to detect the expression and localization of Snail1, SMAD3, SMAD4, α-SMA and E-cadherin in renal tissue. Results Compared with the control group, the mRNA and protein expressions of Snail1, SMAD3, SMAD4 and α-SMA in NRK-52E cells cultured with high glucose and in DKD renal tissues were up-regulated, while E-cadherin expression was down-regulated. After the intervention of VitD3, the expression levels of Snail1, SMAD3, SMAD4, α-SMA and E-cadherin in the DKD model improved to be close to those in the control group. Chromatin immunoprecipitation showed that Snail1 and SMAD3/SMAD4 bound to CAR promoter IV, while VitD3 prevented Snail1 and SMAD3/SMAD4 from binding to CAR promoter IV. Luciferase assay confirmed the interaction among Snail1, SMAD3/SMAD4 and E-cadherin. After the mRNA of Snail1 and SMAD4 was inhibited by siRNA, the expression of E-cadherin induced by high glucose was up-regulated. Conclusion VitD3 could inhibit the formation of Snail1-SMAD3/SMAD4 complex and alleviate the renal tubulointerstitial fibrosis in DKD.

Effect of Yishen Decoction on intestinal microflora and microinflammation for the patients with stage 3-5 chronic kidney disease / 国际中医中药杂志

Objective:To investigate the effects of Yishen Decoction via colonic dialysis on intestinal flora and microinflammation in patients with chronic kidney disease (CKD) stages 3-5. Methods:A total of 156 patients with stage 3-5 CKD from the Second Affiliated Hospital of Guizhou University of Chinese medicine from October 2019 to October 2020 who met the inclusion criteria were divided into 2 groups of 78 patients according to the random number table method. The control group was given colonic dialysis treatment, the treatment group was given Yishen Decoction transcolonic dialysis treatment on the basis of the control group, and both groups were treated for 8 weeks. TCM syndrome scores were performed before and after treatment, serum levels of CRP, IL-6, and TNF-α were measured by ELISA, and SCR, BUN, and uric acid (UA) levels were detected by a fully automated biochemical analyzer. Fresh feces were collected from the patients, anaerobic culture and aerobic culture were performed, and the numbers of Bifidobacterium, Lactobacillus acidophilus, and Escherichia coli were counted to evaluate the clinical efficacy. Results:The total effective rate was 97.4% (76/78) in the treatment group and 84.6% (66/78) in the control group, and the difference was statistically significant ( χ2=7.847, P=0.005). At 4 and 8 weeks after treatment, the scores of lumbar and knee tenderness( t=6.596, 8.792), eating less and being dull ( t=12.060, 24.140) and pale complexion ( t=7.983, 12.300) in the treatment group were significantly lower than those in the control group ( P<0.01); the levels of bifidobacterium ( t=4.037, 2.358) and Lactobacillus acidophilus ( t=7.352, 2.092) were significantly higher than those in the control group, while the levels of Escherichia coli ( t=3.822, 6.084) were significantly lower than those in the control group ( P<0.01 or P<0.05). The serum CRP ( t=9.326, 12.300), IL-6 ( t=4.591, 4.716), TNF-α ( t=9.304, 9.775), SCr ( t=17.630, 11.530), BUN ( t=2.674, 2.248), UA ( t=10.860, 13.160) were significantly lower than those in the control group ( P<0.01 or P<0.05). Conclusion:Yishen Decoction can improve intestinal microecological status, inhibit microinflammatory response and relieve clinical symptoms for the patients with stage 3-5 CKD.

Seminiferous capsule extract effect on the proliferation of spermatogonial stem cells / 中国组织工程研究

BACKGROUND:Chinese herbs for kidney nourishment can promote the proliferation and differentiation of spermatogonial stem cel s. OBJECTIVE:To verify the effect of seminiferous capsule extract on spermatogonial stem cel proliferation. METHODS:Spermatogonial stem cel s were isolated from the testis of male mice and synchronized by serum-free medium fol owed by an addition of 10, 50, 100 mg/L seminiferous capsule extracts. After 24 hours of culture, viability, proliferation and cel cycle of spermatogonial stem cel s were observed. RESULTS AND CONCLUSION:Compared with the control group, seminiferous capsule extracts promoted the cel number, viability and proportion at S stage. The number of BrdU-labeled spermatogonial stem cel s was increased significantly after intervention with seminiferous capsule extracts, especial y at the concentration of 50 mg/L. These findings indicate that seminiferous capsule extracts can promote the proliferation and viability of spermatogonial stem cel s.

Rhodiola polysaccharide effect on spermatogonial stem cell proliferation in vitro / 中国组织工程研究

BACKGROUND:To establish a rapid and effective method to obtain sufficient spermatogonial stem cels that can meet the clinical need is urgent to be solved in the spermatogonial stem cel transplantation. OBJECTIVE:To study the effect of rhodiola polysaccharide on the proliferation of spermatogonial stem celsin vitro. METHODS:Under sterile conditions, spermatogonial stem cels and Sertoli cels were isolated from the testis of mice, and spermatogonial stem cels were seeded onto the feed layer of Sertoli cels. Then, the co-cultured cels were assigned into experimental group 1 (simple cel culture medium), experimental group 2 (cel culture medium containing 150 mg/L rhodiola polysaccharide) and experimental group 3 (cel culture medium containing 150 mg/L rhodiola polysaccharide, 1 U/L leukemia inhibitory factor and 10 μg/L glial cel line-derived neurotrophic factor). After 7 days of co-culture, flow cytometry was used to detect cel proliferation in vitro, and cel viability and positive expression of GFRa-1, Thy-1 and C-kit were calculated. RESULTS AND CONCLUSION:After 7 days of co-culture, the cels grew rapidly and presented with colony and thyrsiform growth, and the number of cel masses increased significantly, al of which were in line with the proliferative features of spermatogonial stem cels. The GFRa-1, Thy-1 and C-kit proteins were expressed in the cel membrane and cytoplasm, mainly in the cel membrane. The viability of spermatogonial stem cels and positive expression of GFRa-1 and Thy-1 were ranked as folows: experimental group 3 > experimental group 2 > experimental group 1, and there were significant differences between groups (P < 0.05). The positive expression of C-kit had no difference between experimental groups 1 and 2, but it was significantly higher in the experimental group 3 than the other two groups (P < 0.05). These findings indicate that rhodiola polysaccharide used alone or combined with leukemia inhibitory factor and glial cel line-derived neurotrophic factor can enhance the proliferative ability of spermatogonial stem celsin vitro.

Clinical Study on Self-made Wuhuang Tang Combined with Western Medicine for Type Ⅲ Prostatitis / 中国药房

OBJECTIVE:To study the action mechanism of the self-made Wuhuang tang(Chinese medicine)in the treatment of type Ⅲ prostatitis(chronic nonbacterial prostatitis,CNBP)so as to promote the action mechanism of traditional Chinese medicine to the molecular level.METHODS:90 patients with CNBP were divided into two groups(n=45 each):one group received western medicine alone,another group received integrated Chinese and western medicine(ICWM)(Wuhuangtang plus western medicine).The indicators such as the expressed prostatic secretions(EPS)routine,IL-1?,and TNF-? before and after the treatment were analyzed statistically.RESULTS:The levels of IL-1? and TNF-? in ICWM-treated group decreased significantly and the clinical symptoms of these patients improved significantly as compared with those treated with western medicine alone.CONCLUSION:The self-made Wuhuang tang can markedly alleviate the symptoms of CNBP patients and decrease levels of IL-1? and TNF-? in patients' EPS,thus it is an effective clinical empirical prescription for CNBP.