OBJECTIVE: To establish an HPLC method for the determination of Quercetin in Duizuoye tablet. METHODS: HPLC analysis was carried out on Dikma-ODS C18 column (250 mm?4.6 mm, 5 ?m) with methanol-0.4% phosphoric acid solution(53∶47) as mobile phase at a flow rate of 1.0 mL?min-1; the detection wavelength was set at 370 nm and the column temperature was maintained at 35 ℃. RESULTS: The calibration curve for Quercetin was linear in the range of 1.31～9.17 ?g?mL-1 (r=0.999 8).The average recovery was 98.06%, with RSD of 1.05% (n=6).CONCLUSION: The adopted method is simple, rapid and accurate, and suitable for the quality control of Duizuoye tablet.

To study the effects of forkhead transcription factor O1 (FoxO1) on the expression of type Ⅳ collagen and desmin in podocytes of diabetic rats.Streptozotocin-induced diabetic rats were divided into three groups:diabetic rats (DM group),rats transfected with blank lentiviral vectors (diabetes mellitus plus LV-pSC-GFP group,LV-NC group),and rats which were transfected with lentiviral vectors carrying constitutively active FoxO1 (diabetes mellitus plus LV-FoxO1-AAA group,LV-CA group).Rats which received an injection of diluent buffer served as normal control.At 2,4,and 8 weeks after transfection,the levels of urine albumin,blood glucose,blood urea nitrogen,and serum creatinine were measured.Realtime PCR and Western blotting were performed to measure the mRNA and protein levels of FoxO1,COL4A3,COL4A5,and desmin in the renal cortex.Moreover,light microscope and electron microscope were used to observe the structural changes in glomerulus and podocytes.Compared with LV-NC and DM group,in LV-CA group,there was a significant increase in the mRNA and protein levels of FoxO1,and a distinct decrease in the levels of urine albumin,blood urea nitrogen and serum creatinine of rats (except at the twoweek time point) (all P＜0.05),the mRNA and protein levels of COL4A3,COL4A5,and desmin were all decreased (all P＜0.05),and pathological changes in kidney were also improved.Upregulating the expression of FoxO1 by transfecting with constructed lentiviral vectors can definitely improve the abnormal expression of type Ⅳ collagen and desmin in podocytes of diabetic rats.

Objective To study the role and molecular mechanism of forkhead transcription factor O1 (FoxO1) on proliferation of mesangial cells( MCs) in diabetic rats. Methods Empty lentiviral vector( LV-pSC-GFP) and the constitutively active FoxO1 lentiviral vector(LV-CA-FoxO1) were constructed. Diabetic rat model was established and rats were divided into diabetes group(DM group), diabetes with LV-pSC-GFP group(NC group), and diabetes with LV-CA-FoxO1 group(CA group). The normal SD rats of the same age were considered as the normal control group(NG group). The lentiviral vector was injected into the renal cortex of diabetic rats in corresponding groups. Body weight, blood glucose, 24 h urinary protein, urine albumin, serum creatinine, and blood urea nitrogen was detected at the end of 2 weeks, 4 weeks, and 8 weeks. The ratio of kidney weight/ body weight was counted and the renal cortex was reserved for light microscopy, electron microscopy and frozen section after rats were sacrificed in different groups. The mRNA level of FoxO1 and p27Kip1 were detected by real-time PCR. The protein expressions of FoxO1, p-FoxO1, and p27Kip1 were tested by Western blotting. Results The renal pathological changes were obviously ameliorated in CA group. Compared with DM group, the mRNA and protein expression of FoxO1 and p27Kip1 were significantly increased in CA group (P<0. 05), whereas there was no difference in the expression of p-FoxO1 protein(P > 0. 05). The p-FoxO1 / FoxO1 ratio was decreased ( P < 0. 05). All indexes had not reached statistical difference between NC group and DM group(P>0. 05). Conclusion Overexpression of FoxO1 in kidneys of diabetic rats can inhibit the proliferation of mesangial cells, and may through up-regulating the expression of p27Kip1 delay the progression of diabetic nephropathy.

Objective To analyze the clinical manifestations and possible gene mutation sites of Chinese patients in order to improve the clinician's understanding of CHARGE syndrome. Methods Clinical data were collected and blood samples were obtained from the proband of CHARGE syndrome and their relatives. The peripheral blood DNA was extracted and sequenced by PCR amplification. Mutation sites were verified by Sanger sequencing. Results For the first proband, a heterozygous mutation was detected in the intron 10 of CHD7 gene. His parents and brother did not have mutation. For the second proband, total repeat sequence in exon 7 of CHD7 gene was detected. His father carried the same mutation and his mother did not have mutation. Conclusion For the patients who are diagnosed with CHARGE syndrome based on the clinical manifestations, genetic mutation detection should be proceeded. It is useful for studying possible genetic pathogenesis and enhancing the awareness of clinicians.

Objective:To study the expression of peripheral blood NKT-like cells in patients with systemic sclerosis (SSc), to explore the correlation between NKT-like cells and laboratory and clinical indicators of systemic sclerosis, and investigate the role of NKT-like cells in the occurrence and development of Systemic sclerosis.Methods:Forty-six SSc patients (SSc group) were enrolled from Department of Rheumatology and Immunology of Peking University People 's Hospital during December 2018 to December 2019. Thirty healthy subjects with matched age and sex were selected as healthy control group (HC group). The cell count and percentage of NKT-like cells and other lymphocyte subsets in peripheral blood were detected by flow cytometry. At the same time, other laboratory indexes were determined by different methods. Spearman's correlation analysis, Pearson's correlation analysis, Man-Whitney U test and Fisher's exact test were used to analyze the difference and correlation between NKT-like cells and other clinical and laboratory indicators. Results:Compared with HC group [165(72, 226)cells/μl], the cell count of NKT-like cells in peripheral blood of SSc group[30(19, 58)cells/μl] was significantly decreased ( Z=-5.69, P<0.001). Correlation analysis showed that the cell count of NKT-like cells was positively correlated with total T lymphocytes ( r=0.56, P<0.001), CD4 +T cells ( r=0.42, P=0.004), CD8 +T cells ( r=0.60, P<0.001), B cells ( r=0.50, P<0.001) and NK cells ( r=0.33, P=0.024), respectively. The percentage of NKT-like cells in lymphocytes was also positively correlated with the percentage of CD8 +T cells ( r=0.34, P=0.020), but not significantly correlated with other subset of lymphocytes. The ESR of the NKT-like cell decreased group was significantly higher than that of the NKT-like normal group[15(9, 28) mm/1 h vs 8 (4, 16) mm/1 h, Z=-2.04, P=0.042]. Moreover, the cell count of NKT-like cells was negatively correlated with ESR ( r=-0.34, P=0.019). Conclusion:The cell count and percentage of NKT-like cells in peripheral blood of SSc patients decreased significantly. NKT-like cells were not only positively correlated with a variety of lymphocyte subpopulations, but also negatively correlated with ESR. NKT-like cells may be used as an indicator to monitor the disease activity in patients with SSc.