OBJECTIVE:To establish a method for the simultaneous determination of flavonoids components in Astragali Ra-dix,and to explore the relationship among flavonoids components,varieties,origins and planting patterns. METHODS:HPLC was performed on the column of Venusil ASB with mobile phase of acetonitrile-0.3% formic acid (gradient elution) at a flow rate of 1.0 ml/min,detection wavelength was 260 nm,and column temperature was 25 ℃. Medicinal material quality of Astragalus mem-branaceus (Fisch.) Bge. var. mongholicus (Bge.) Hsiao and A. membranaceus (Fisch.) Bge of wild and cultivated from different province was compared. RESULTS:The linear range of the mass concentration was 0.008 9-2.224 mg/ml for calycosin glucoside (r=0.999 5),0.005 2-1.3 mg/ml for ononin(r=0.999 6),0.002 8-0.697 6 mg/ml for calycosin(r=0.999 9)and 0.002-0.5 mg/ml for formononetin (r=0.999 9);RSDs of precision,stability and reproducibility tests were lower than 1%;recoveries were 99.52%-100.74%(RSD=0.41%,n=6)for calycosin glucoside,98.84%-100.60%(RSD=0.60%,n=6)for ononin ,98.47%-101.74%(RSD=1.08%,n=6)for calycosin,100.10%-101.59%(RSD=0.32%,n=6)for formononetin. In terms of varieties,the contents of calycosin glycosides,ononin and flavonoids in A. membranaceus(Fisch.)Bge. var. mongholicus(Bge.)Hsiao were higher than those of A. membranaceus (Fisch.)Bge,but the contents of calycosin and formononetin were less than those of A. membranaceus (Fisch.)Bge;in terms of origins,calycosin glycosides and flavonoids of Inner Mongolia and Shanxi held the highest contents,fol-lowed by those of Northeast China and Gansu,and lowest in Shandong,Anhui and Shaanxi;in terms of planting patterns,the con-tents of calycosin glycosides,ononin and flavonoids of wild Astragali Radix were higher than those of cultivated varieties,and the contents of calycosin and formononetin of cultivated varieties were higher than those of wild ones. CONCLUSIONS:The method is simple,stable and reproducible,and can be used for the simultaneous determination of flavonoids components in Astragali Radix. The flavonoids components show great differences in Astragali Radix from different origins,and they are affected by varieties,ori-gins and planting patterns.

Objective To evaluate the protective effects of Rosemary's chloroform extract on cerebral ischemia in mice and acute toxicity.Methods The protective of rosemary's chloroform extract on cerebral ischemia was observed and compared by the mice models of cerebral anoxia and ischemia,thrombus formation in vivo,and chloroform induced arrhythmias.Rosemary's Chloroform Extract was given orally to mice to evaluate acute toxicity.Results Rosemary's chloroform extract had different degrees of inhibition of collagen-the adrenaline induced thrombus formation,and prolonged acute ischemic mice brains off mouth breathing time and increase the number of mouth breathing (P ＜0.05 or P ＜0.01);Chloroform extract significantly reduced the incidence of chloroform induced ventricular arrhythmias.Acute toxicity results suggested that a female rat died in the next day of chloroform extract group,no additional toxicity was observed.Conclusion Rosemary's chloroforrm extract has significant protective effect on cerebral ischemia and hypoxia in mice.

Objective: To clarify the relationship between the hemoglobin level and renal tubular atrophy/interstitial fibrosis (T) in the Oxford stage of renal pathology in IgA nephropathy (IgAN) patients. Methods: Patients diagnosed with IgAN by renal biopsy from January 1^st 2010 to December 31^st 2015 in Shenzhen Second People's Hospital with complete laboratory and imaging data were retrospectively analyzed. Patients were divided into anemic group and non-anemic group. The relationship between hemoglobin level and renal tubular atrophy/interstitial fibrosis was determined by logistic regression analysis. The possible curve relationship between hemoglobin and renal tubular atrophy/interstitial fibrosis was analyzed by smooth curve fitting analysis. The diagnostic value of hemoglobin for renal tubular atrophy/interstitial fibrosis was analyzed by the receiver operating curve (ROC). Results: A total of 630 patients with IgAN were included in this study, 130 patients in the anemia group (20.63%) and 500 patients in the non-anemia group (79.37%). There was no statistically significant difference in age between the two groups, but the difference of the gender was statistically significant (male 35.38% vs 53.80%, χ²=10.740, P<0.001). Compared with the non-anemia group, the anemia group had a higher proportion of tubular atrophy/interstitial fibrosis (χ²=62.586, P<0.001), higher 24 h urinary protein quantification (Z=-6.082, P<0.001), and lower eGFR (t=7.126, P<0.001). Multivariate logistic regression analysis showed that increasing hemoglobin level was an independent protective factor for reducing the risk of renal tubular atrophy/interstitial fibrosis (OR=0.973, 95%CI 0.958-0.987, P<0.001). Smooth curve fitting analysis showed that there was a linear negative correlation between hemoglobin and tubular atrophy/interstitial fibrosis. The ROC curve suggested that the best threshold of hemoglobin was 120.5 g/L when renal tubular atrophy/interstitial fibrosis occurred. That was, when hemoglobin was above 120.5 g/L, the severity level of renal tubular atrophy interstitial fibrosis might be reduced. Conclusion The incidence of renal tubular atrophy/interstitial fibrosis is higher in IgAN patients with anemia, and hemoglobin>120.5 g/L may reduce the risk of tubular atrophy/interstitial fibrosis.

Objective To explore the clinicopathological features and outcomes of IgA nephropathy patients with different proportions of crescents.Methods A total of 270 patients who were diagnosed as IgA nephropathy by renal biopsies from January 2010 to December 2015 in the First Affiliated Hospital of Shenzhen University were enrolled.All patients were divided into 3 groups according to the optimal cutoff level of crescents proportion in the Receiver Operating Characteristic Curve (ROC) as follows:0％,＜ 14％;≥ 14％.The endpoint was defined as the doubling of baseline serum creatinine (Scr) and/or end-stage renal disease (ESRD).Kaplan-Meier curve and Cox regression model were used to analyze the renal survival among three groups.Results One hundred and four patients (38.5％) without any crescents;84 patients (31.1％) with crescents proportion ＜ 14％ and 82 patients (31.4％) with crescents proportion ≥14％.Patients with crescents proportion ≥14％ group were older and had higher level of systolic blood pressure and diastolic blood pressure,24-hour urine protein and serum uric acid level;more patients treated with RAS blocker,glucocorticoid and immunotherapy,but lower eGFR,hemoglobin and serum albumin level than those with crescents proportion ＜ 14％.Compared with those without crescents and crescent proportion ＜ 14％,patients with crescent proportion ≥ 14％ also had higher proportion of global glomerulosclerosis,more endocapillary hypercellularity and severe tubulointerstitial lesions,higher degree of IgA and C3 depositions in renal.24-hour proteinuria,serum uric acid level,low hemoglobin level,endocapillary hypercellularity and renal C3 depositions were risk factors for crescents formation.Patients were followed-up for a median of (31.7±21.0) months,and Kaplan-Meier analysis revealed that renal survival rate was significantly lower in patients with crescents proportion ≥ 14％ compared with other groups (P=0.001).But there was no significant difference between no crescent group and crescents proportion ＜ 14％ group.However,multivariate Cox analysis showed no significant difference between crescents proportion and renal survival.Conclusion Crescents proportion is associated with higher risk of renal function and renal progression.

Objective To investigate the relationship between interventricular septum thickness (IVST) and renal prognosis in IgA nephropathy patients.Methods A total of 213 patients with IgA nephropathy proven by biopsy from Department of Nephrology of Shenzhen Second People's Hospital were enrolled in this study,and these participants were divided into normal IVST group (＜ 11 mm) and higher IVST (≥ 11 mm) group according to IVST.The demographic characteristics,clinical biochemical indexs,CKD stage and pathologic characteristics in these two groups were compared.Binary logistic regression analysis was used to analyze the influencing factors of eGFR ＜ 60 ml · min-1 ·(1.73 m2)-1,and Kaplan-Meier survival curve was used to analyze the effect of IVST on renal prognosis.Results Compared with IVST normal group,the patients in IVST higher group were more male sex,older,and had higher level of systolic pressure,Hb,Scr,BUN,UA,24 h urine protein excretion,urinary protein creatinine ratio,triacylglycerol,total cholesterol,LDL,Serum C3,C4,and had more serious mesangial proliferation,tubular atrophy (all P ＜ 0.05).However,the levels of eGFR and HDL were decreased in IVST higher group (both P ＜ 0.05).There were a significant difference in CKD staging distributions and IgA Lee grade between two groups (both P ＜ 0.01).Spearman and Pearson correlation analysis indicated that IVST was negatively correlated with eGFR and positively correlated with proteinuria level in IgA nephropathy patients.Baseline IVST was an independent risk factor of eGFR ＜ 60 ml · min-1 · (1.73 m2)-1 in IgA nephropathy patients.Serum C3,UA and hemoglobin were independent influential factors of eGFR decline (all P ＜ 0.05).Kaplan-Meier survival curve indicated that the renal function was worse in patients with thickened interventricular septum.Conclusion The IgA nephropathy patients with thicker interventricular septum has a poor renal prognosis.

Objective To investigate the protective effects of stilbene glycoside(2,3,5,4'-tetrahydroxy-stilbene-2-O-β-D-glucoside,TSG) on 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced mice model of Parkinson's disease(PD).Methods Mice were randomly divided into the blank control group,the negative control group,the TSG high-dose group,the TSG low-dose group and the positive drug group(n=20 each).Mice were weighted daily to observe the changes of body weight,and mice motor and behavior function were tested by open field test.Level changes of α/β synuclein in brain cortex,cerebellum,midbrain,and hippocampal were detected by Western blot.Results Compared with the blank control group,the negative control group showed that the body weight was decreased (P ＜ 0.05).Compared with the negative control group,the body weight was increased in the TSG high-and low-dose groups and the positive drug group (P ＜ 0.05).The spontaneous behavior was impaired in the negative control group.Compared with the blank control group,the negative control group showed that the open field test showed traveled distance over a 10-min period was significantly shortened at 1 st,7th,28th days after testing(all P＜0.05).The trajectory of motor axons indicated that mice in the negative control group showed dyskinesia,but the groups of positive drug and high-and low-dose of TSG could reverse this dyskinesia.Compared with the blank control group,brain α/β synuclein protein levels were increased in the negative control group,and decreased in positive drug and TSG high-and low-dose groups (P ＜0.05).Conclusions Stilbene glycosides exert neuroprotective effects in MPTP-induced mice model of PD.

Objective:To investigate the relationship between hemoglobin levels and renal prognosis in patients with IgA nephropathy (IgAN).Methods:The clinical data and pathological examination results of IgAN patients diagnosed by renal biopsy at the Second People's Hospital of Shenzhen from February 25, 2010 to September 9, 2020 were retrospectively collected and analyzed. The patients were divided into anemic group and non-anemic group according to the anemia diagnostic criteria (The hemoglobin levels were<120 g/L and<110 g/L in males and females respectively at sea level area). Endpoint event was defined as a decrease in estimated glomerular filtration rate (eGFR) of>50% from baseline and/or progression to stage 5 chronic kidney disease [eGFR<15 ml·min -1·(1.73 m 2) -1]. Cox regression analysis was used to analyze the factors affecting the poor renal prognosis. The relationship between hemoglobin and renal function prognosis was analyzed by smoothing curve fitting and threshold effect. Kaplan-Meier survival curve was used to compare and analyze the difference of renal survival rate between the anemic and non-anemic IgAN patients. Results:A total of 1 263 IgAN patients were included in this study, 255(20.19%) patients were in the anemia group and 1 008 (79.81%) patients were in the non-anemia group. The anemia group had lower body mass index, baseline eGFR, serum albumin, and triglyceride than those in the non-anemia group (all P<0.05). The proportion of females, 24 h urinary protein content, and the proportion of renal tubule atrophy/interstitial fibrosis, segmental sclerosis and crescents in the anemia group were higher than those in the non-anemia group (all P<0.05). Multivariate Cox regression analysis showed that low hemoglobin was an independent influencing factor for renal endpoint event ( HR=0.25, 95% CI 0.07-0.90, P=0.022). Smoothing curve fitting analysis and threshold effect analysis showed that a curving relationship was detected between hemoglobin and relative risk of renal endpoint event. As hemoglobin increased, there was a protective effect on renal function when hemoglobin level was lower than 147 g/L ( β=0.96, 95% CI 0.94-0.99, P=0.008). Kaplan-Meier survival curve analysis suggested that patients with anemia had a lower cumulative renal survival rate than that of patients without anemia (Log-rank test χ2=10.106, P=0.002). Conclusions:Low hemoglobin is an independent influencing factor for poor prognosis of renal function in IgAN patients. Cumulative renal survival rate is lower in IgAN patients with anemia than that of patients without anemia.

Objective:To explore the correlation between hemoglobin (Hb) and progression of renal function in patients with proliferative lupus nephritis (PLN).Methods:Data of biopsy-proven PLN patients from January 2010 to February 2019 in Department of Nephrology, the First Affiliated Hospital of Shenzhen University were retrospectively analyzed. The patients were divided into stable renal function group and renal function progression group according to changes of renal function including serum creatinine doubling/end-stage renal disease (ESRD) and the demographic, clinical, and pathological characteristics were compared between the two groups. Cox regression and smooth curve fitting of generalized additive model analysis were used to explore the correlation between Hb and progression of renal function.Results:A total of 87 patients were included in this study. The age was (34.97±11.95) years old and 79 cases (90.80%) were females. During follow-up of 19.0(6.5, 43.5) months, renal function of 15 patients (17.24%) progressed. Compared with stable renal function group, Hb level of patients in renal function progression group were lower ( t=3.887, P<0.001), and serum creatinine ( Z=-2.466, P=0.003) and uric acid ( t=-2.154, P=0.034) were higher. As to the pathological characteristics, the proportion of class lupus nephritis-IV, renal tubular atrophy and interstitial fibrosis in renal function progression group were higher than those in stable renal function group, but there was no statistical difference between the two groups (all P>0.05). Multivariate Cox regression analysis indicated that high Hb was an independent protective factor of renal function progression in PLN patients ( HR=0.893, 95% CI 0.836-0.954, P=0.001). The risk of progression to serum creatinine doubling/ESRD would decrease by 10.7% when Hb increased by 1 unit (g/L). Smooth curve fitting of generalized additive model analysis showed that Hb was linearly correlated with the risk of renal function progression ( P=0.100). Receiver operating characteristic curve indicated that the risk of doubling serum creatinine/ESRD in PLN patients would be relatively low when Hb level was above 77 g/L (area under the curve 0.788, best threshold 77 g/L, sensitivity 0.600, specificity 0.903). Conclusions:Hb is closely related to progression of renal function in patients with PLN. More attention and management of Hb levels in patients with PLN can play an important role in improving renal prognosis.

Objective:To investigate the correlation between serum C3 and glomerular microthrombosis in patients with lupus nephritis (LN).Methods:Patients who were diagnosed as LN by renal biopsy hospitalized in Department of Nephrology, the First Affiliated Hospital of Shenzhen University from January 2010 to February 2019 were retrospectively analyzed and they were divided into glomerular microthrombosis group (GMT group) and non-glomerular microthrombosis group (non-GMT group). The demographic data, clinical characteristics, pathology and prognosis of the two groups were compared. Logistic regression and smooth curve fitting of generalized additive mixed model analysis were used to explore the correlation between serum C3 and glomerular microthrombosis. Renal prognosis of the two groups were compared by the Kaplan-Meier survival curve.Results:A total of 116 patients were enrolled, aged (32.79±11.43) years old, in which 108 cases (93.10%) were female. Thirty-seven patients (31.90%) were confirmed to be combined with GMT (GMT group) and 79 cases were not (non-GMT group). Compared with the non-GMT group, patients in the GMT group were relatively older ( t=-2.876, P=0.002), with higher proportion of hypertension ( χ2=7.492, P=0.006), higher urine protein quantitation ( Z=-2.115, P=0.003), lower levels of eGFR and serum complement C3 ( Z=3.469, P<0.001; t=1.744, P<0.001), higher systemic lupus erythematosus disease activity index ( t=-2.758, P=0.007). As to the pathological characteristics, type IV LN patients were the majority (72.97%). Proportion of crescents and pathological activity indicators of the GMT group were higher ( Z=-1.866, P=0.002; t=-5.005, P<0.001), nuclear fragmentation, endothelial hyperplasia and renal tubular atrophy were more serious ( χ2=14.987, P<0.001; χ2=15.695, P<0.001; χ2=4.130, P=0.042). Multivariate logistic regression analysis indicated that serum complement C3 was a relational factor of the formation of GMT in LN patients ( OR=0.966, 95% CI 0.938-0.995, P=0.023). Smooth curve fitting of generalized additive mixed model analysis indicated that level of complement C3 had a linear relationship with the changing trend of GMT. The Kaplan-Meier curve showed that there were statistical differences between the two groups in terms of complete remission of urine protein (Log-rank χ2=5.858, P=0.016) and doubled serum creatinine/end-stage renal disease (Log-rank χ2=3.945, P=0.047). Conclusions:Serum C3 is closely related to the formation of GMT in LN patients, and statistical differences were demonstrated in the renal prognosis of GMT group and non-GMT group.

Objective:To establish a diagnostic model for glomerular micro thrombosis (GMT) in lupus nephritis through clinical indicators.Methods:A continuous collection of patients diagnosed with lupus nephritis (LN) by renal biopsy in the Department of Nephrology, Shenzhen Second People's Hospital, from January 2010 to March 2021. All patients were admitted and discharged through the inclusion and exclusion criteria. Demographic data, clinical characteristics, biochemical indicators, and immune indicators were collected. A GMT diagnosis model was established from the most important variables among the abovementioned variables through machine learning and Logistic stepwise regression analysis. The model was presented through a nomogram. The receiver operating characteristic curve (ROC), the clinical decision curve and the calibration curve were used to evaluate the model discrimination, clinical use and accuracy, respectively. The internal verification of the model was carried out by repeated sampling 500 times by the Bootstrap method.Results:There were a total of 129 patients with lupus nephritis including the study, including 117 females (90.7%); the average age was (34±11) years. There were 39 patients with GMT (30.2%). Using machine learning to screen out the top 10 important variables from 47 candidate variables, then through logistic stepwise regression analysis, five variables were further screened to establish the diagnostic model of GMT, namely hemoglobin [ OR(95% CI)=0.966(0.943, 0.990), P=0.005], serum C3 [ OR(95% CI)=0.133(0.022, 0.819), P=0.030], systolic blood pressure [ OR(95% CI)=1.027(1.005, 1.049), P=0.017], lymphocyte count [ OR(95% CI)=0.462(0.213, 0.999), P=0.049], and TT [ OR(95% CI)=1.260(0.993, 1.597), P=0.057]. Draw up the equation of the GMT diagnosis model of lupus nephritis and establish a nomogram to present the model. The area under curve (AUC) of the diagnostic model was 0.823, 95% CI(0.753, 0.893), indicating that the model had a reasonable degree of discrimin-ation. The Hosmer-Lemeshow test showed a perfect fit between the predicted GMT risk and the observed GMT risk ( χ2= 14.62, P=0.067). The clinical decision curve and clinical impact curve reflect that the model had a good clinical application value, especially when the threshold probability is between 0.4 and 0.6, the application value is more significant. In addition, after 500 repeated samplings by the Bootstrap method, the average AUC was 0.825, 95% CI(0.753, 0.893), which is basically the same as the AUC obtained by the original model. Conclusion:We established a diagnostic model of GMT inLN based on clinical indicators through machine learning and logistic stepwise regression analysis. It is used for early diagnosis of the risk of GMT before the renal biopsy.