BACKGROUND: Hepatic fibrosis is a reversible disease, interfering in course of disease promptly can decrease hepatic cirrhosis and fatal complication. Hepatic stellate cells (HSCs) is a key factor in pathogenesy of hepatic fibrosis.OBJECTIVE: To investigate the effects of Shugan granule on HSCs activation and trans-membrane signal transduction stimulated by transforming growth factor beta 1 (TGF-β1) in rats, and to explore the anti-fibrosis mechanism.DESIGN, TIME AND SETTING: Controlled observational trials based on cytology were performed in the Central Laboratory of Molecules, Luzhou Medical College between June 2008 and February 2009. MATERIALS: HSC-T6 cell line was purchased from Institute of Liver Diseases, Shanghai University of Traditional Chinese Medicine, its phenotype was the activated hepatic stellate cells. Shugan granule was offered by Drug Institute, Affiliated Hospital of Luzhou Medical College at a Batch No. 20071120.METHODS: The influence of different concentrations (0.01, 0.02, 0.04, 0.08 g/L) of Shugan granule on HSCs proliferation was determined by MTT. 0.01 g/L was defined as the dose of Shugan granule contributing no influence on cell proliferation. HSCs were cultured in a culture plate and then divided into 4 groups: control group without management, TGF-β1 group with 5 μg/L TGF-β1 solution in culture medium, Shugan granule group with 0.01 g/L Shugan granule in a culture medium, and TGF-β1 + Shugan granule group with 5 μg/L TGF-β1 solution and 0.01 g/L Shugan granule in culture medium. MAIN OUTCOME MEASURES: Morphological features of HSCs were detected by microscopic. Immunohistochemical method was used to detect the expression of Smad3 and Smad7 in HSCs. RT-PCR was applied to observe the HSCs activation and trans-membrane signal transduction. RESULTS: ①The cell morphology of TGF-β1 group was similar with that in the control group, and the extension was more obvious. In the TGF-β1 + Shugan granule group, the cell morphology was close to that in TGF-β1 group. There was no karyopyknosis or apoptosis observed in each group. ②Immunohistochemical method showed the expression of Smad3 and Smad7 in control groups were increased. TGF-β1 could slightly increase the expression of Smad3 and Smad7 (P < 0.05), while Shugan granule group and TGF-β1 + Shugan granule group increased the expression of Smad7 significantly, accounting for 1.99 times compared with control group (P < 0.01). ③RT-PCR result showed that Shugan granule could increase the expression of Smad7 (P < 0.05), but the expression of Smad3 was not regulated. 5 μg/L TGF-β1 could up-regulate the expression of Smad3 and Smad7 (P < 0.05). In the TGF-β1 + Shugan granule group, Smad7 expression was increased by 101% (P < 0.05), but Smad3 transcriptional level was not changed(P > 0.05). CONCLUSION: ①TGF-β can stimulate the gene expression of Smad3 and Smad7, it also obtain a balance of feedback regulation mechanism between R-Smads and I-Smads. ②Shugan granule may prevent and cure hepatic fibrosis through decreasing the proliferation of HSCs in a dose-dependent manner. ③Shugan granule can inhibit the TGF-β-Smad signaling pathway through increasing the expression of Smad7.

Objective:To evaluate the expression of IMP3 and CD44 proteins in recurrent urothelial carcinoma (UC) and to deter-mine the correlation between the two proteins. Methods: Data from transurethral resection of bladder (TURB) cancer cases between January 2002 and December 2012 were reviewed. Of the 54 UC recurrent cases in this study, one group of 25 had experienced recur-rence within 6 months after surgery, and the other group of 29 had their first recurrence after more than 3 years. IMP3 and CD44 immu-noreactivities were increased, which correlated with the clinicopathologic parameters. The relationship between IMP3 and CD44 pro-tein expressions was also explored. Results:Six of the 25 short-term recurrent UC cases were tested positive for IMP3 and all belonged to high-grade UC. Among the 29 long-term recurrent patients, only one case of low-grade UC tested positive for IMP3. IMP3 expres-sion rate [24%(6/25)] and intensity [weak staining at 16%(4/25) and strong staining at 8%(2/25)] were higher in the short-term recur-rent group than those in the long-term group, which had an expression rate of 3.45% (1/29) and intensity rates for weak staining at 3.45%(1/29) and without strong staining (0/29). No difference was observed in the CD44 expression between the two groups. In addi-tion, the high expression of IMP3 correlated with higher tumor stage and grade, whereas the CD44 expression tended to be inversely correlated with the tumor grade in recurrent UC patients. Furthermore, no correlation existed between the expression of IMP3 and CD44 proteins in the bladder carcinoma specimens. Conclusion:IMP3 exhibited a significantly higher expression rate in short-term re-current UC specimens than in the long-term recurrent cases. Therefore, IMP3 could be used as a novel marker, together with the other factors including tumor stage and grade, for predicting the high risk of short-term recurrence in UC patients who underwent TURB.

This study was aimed to observe the influence ofQing-Chang Hua-Shi Recipe (QHR) on IL-6trans-signaling in experimental colitis mice, in order to initially explore the possible mechanisms of QHR for ulcerative colitis (UC). TNBS/ethanol was used in the establishment of colitis mice model. After intervention of medication, ELISA was used in the detection of soluble Interleukin-6 receptor (sIL-6R). Real-time PCR was used to detect the mRNA expression level of IL-6 and glycoprotein 130 (gp130). Western blot was used in the observation of protein expression of IL-6 and gp130 in the colonic mucosa. The results showed that the level of sIL-6R, the mRNA and protein expression of IL-6 and gp130 in the model group were significantly higher than that in the control group. QHR was able to reduce the sIL-6R level (P < 0.01), decreased the mRNA and protein expression of IL-6 and gp130 (P < 0.01) in the colon tissues among experimental colitis mice. It was concluded that QHR had good anti-inflammatory effects on experimental colitis mice. It might be associated with influencing IL-6trans-signaling.

Aim To investigate the proliferative effect and the apoptosis of human hepatoma SMMC-7721 cells induced by gallic acid ( GA ) , and its underlying mechanism. Methods SMMC-7721 cells were cul-tured in vitro. MTT assay was used to observe the pro-liferation of SMMC-7721 cells induced on GA 24 , 48 , 72 h. The morphological and ultra structural changes of the SMMC-7721 cells were observed by inverted micro-scope and transmission electron microscope respective-ly. Annexin V-FITC/PI staining was used to quantify the percentages of apoptosis in the total cell popula-tion. The expression of p53 mRNA was investigated by RT-PCR. Western blot was used to determine the pro-tein expression of p53. Results GA(6. 25~50 μmol ·L-1 ) markedly inhibited the activity of proliferation and induced apoptosis of SMMC-7721 cells after 48 h in a dose-dependent manner. GA significantly induced cell nuclear condensation and fragmentation. RT-PCR and Western blot results showed that GA could improve the expression of p53 mRNA and protein. Conclusion GA can inhibit the proliferation of human hepatoma SMMC-7721 cells and induce cells apoptosis. The mechanism may be associated with improving tumor suppressor gene p53 expression.

BACKGROUND: Continuous blood purification can remove cytokines and inflammatory mediators, maintain homeostasis and prevent the occurrence of multiple organ dysfunction syndrome in patients with severe pancreatitis, which has become the main therapy for severe pancreatitis. Since the hemodialysis technology began to be applied clinical y, the biological and physicochemical properties of hemodialysis membrane materials have been studied. A variety of hemodialysis membranes have been developed in order to improve the biocompatibility and anticoagulant effect in vitro. OBJECTIVE: To investigate the application effect of hemodialysis membranes on severe pancreatitis. METHODS: Ten Wistar rats were selected to make rat models of severe pancreatitis and then were randomized into two groups (n=5 per group): homophone membrane group and polysulfone membrane group. Hemodialysis- related biochemical parameters were detected in the two groups. RESULTS AND CONCLUSION: Compared with the homophone membrane, ultrafiltration coefficient, creatinine clearance, blood urea nitrogen clearance, phosphorus clearance, number of circulating endothelial cel s, and levels of plasma nitric oxide and asymmetric dimethylarginine were significantly lower in the polysulfone membrane group (P < 0.05). Vitamin B12 clearance and amount of pre-congestion increased significantly in the polysulfone membrane group as compared with the homophone membrane (P < 0.05). These findings indicate that the polysulfone membrane for hemodialysis has good biocompatibility, and keeps a stable environment in vivo for severe pancreatitis patients.

Aim To study the synergistic anti-depres-sion effect of 3 , 6-disinapoyl sucrose ( DISS ) and tenuifoliside A ( TFSA ) from Radix Polygalae and the preliminary mechanism . Methods Using the classical behavioral despair and depression model of mouse tail suspension test, 120 mice were divided into control group, positive group, DISS 5 mg·kg-1 group,DISS 10 mg·kg-1 group,TFSA 5 mg·kg-1 group,TFSA 10 mg· kg-1 group, DISS 5 mg · kg-1 +TFSA 5 mg · kg-1 group,DISS 5 mg·kg-1 +TFSA 10 mg·kg-1 group,DISS 10 mg·kg-1 +TFSA 5 mg·kg-1 group and DISS 10 mg · kg-1 +TFSA 10 mg · kg-1 group randomly. They were given intragastric injection for 7 days continuously, to observe the effect of DISS and TFSA monomer and its combination on the time of mouse tail suspension. Expression of BDNF in the hip-pocampus of mice was detected by immunohistochemis-try. The expressions of CREB, pCREB, CRTC1 and BDNF in the hippocampus of mice were detected by Western blot method. Results The administration of DISS and TFSA could shorten the immobility time of mice subjected to the tail. DISS ( 10 mg · kg-1 ) and TFSA( 10 mg · kg-1 ) group were significantly lower than single dose drug group(P<0. 05). DISS and TF-SA and the combination groups could increase the ex-pression of BDNF in hippocampus and cortex by immu-nohistochemistry(P <0. 05). At the same time, the contents of CREB, CRTC1, pCREB and BDNF protein in the hippocampus were increased by DISS and TF-SA, and the combination group was significantly higher than the single drug group ( P<0. 05 ) . Conclusion The administration of DISS and TFSA are used to acti-vate CREB transcription factor CRTC1 , and activate the phosphorylation of CREB in the hippocampus, and then increase the expression of BDNF in the hippocam-pus and plays a synergistic antidepressant effect.

OBJECTIVE: To explore the clinical and genetic characteristics of a Chinese pedigree affected by glycogen storage disease (GSD) type Ia with gout as the first manifestation. METHODS: Clinical and biochemical data of the pedigree were collected. Available members of the pedigree were subjected to gene sequencing, and the result was analyzed by bioinformatics software. The pedigree was followed up for five years. RESULTS: The proband was a young female manifesting recurrent gout flare, hypoglycemia, and hypertriglyceridemia. One of her younger brothers also presented with dysplasia and hepatic adenoma. Gene sequencing revealed that the proband and her younger brother both harbored c.1022T>A (p.I1e341Asn) and c.230+5G>A compound heterozygous variants of the G6PC gene , which were inherited from their father and mother, respectively. Among these, the c.230+5G>A is an intron region variant which was unreported previously, and bioinformatics analysis showed that it may impact mRNA splicing of the gene. The proband was treated with raw corn starch, allopurinol, and fenofibrate. Gout was well controlled, and she had given birth to a baby girl without GSD. CONCLUSION GSD Ia should be considered among young gout patients with hypoglycemia and hepatomegaly, for which gene sequencing is warranted. GSD Ia has a good prognosis after comprehensive treatment with diet and medicine.
China ; Female ; Glycogen Storage Disease Type I ; Gout/genetics* ; Humans ; Hypoglycemia ; Male ; Pedigree ; Symptom Flare Up

OBJECTIVE To investigate the activation of xanthine oxidase(XO)from the human liver by vitamin K3 and the mechanism.METHODS Using human liver S9(0.1 g·L-1)as the source,XO was incubated with substrate xanthine of 0,2,4,8,and 16 μmol·L-1 at 37℃ for 90 min.The Michaelis constant(Km)of the reaction of xanthine oxidation was determined using the liquid chromatography diode array method.At the concentration of Km,the three-point method(1,10 and 100 μmol·L-1)was used to detect the activity of vitamin K3 activators.The multi-point method(vitamin K3 1,2,5,10,20,50,100,200 and 400 μmol·L-1)was adopted to determine the half effective concentration(EC50)of activated XO.Kinetic parameters(Km and Vmax)and the fit of double reciprocal curves were determined via vitamin K3 of 1/2EC50,EC50 and 2EC50.The changes in kinetic behavior at different concentrations of vitamin K3 were observed and their types of activation were analyzed.The interactions between XO and activator vitamin K3 were explored via molecular docking.RESULTS The Km of XO-mediated xanthine oxidation reac-tion was 4.71 μmol·L-1.As an activator of this reaction,vitamin K3 activated XO in a concentration-dependent manner(according to the logistic fitting formula y=A2+(A1-A2)/(1+(x/x0)^p),with an EC50 of 32.0 μmol·L-1.The kinetic parameters also changed after the addition of vitamin K3.The Km value decreased(4.71-1.34 μmol·L-1)with the increase of vitamin K3 concentrations,while the Vmax value increased(0.08-1.31 μmol·min-1·g-1),leading to an increase in Vmax/Km(17.0-977.6 mL·min·g-1).In addition,the double reciprocal curve fitting found that the activation type of vitamin K3 on XO was mixed.The molecular docking results showed that vitamin K3 bound to the molybdopterin domain of XO and maintained hydrogen bonding interactions with Arg599 and Ser605.CONCLUSION Vitamin K3 is an activator of XO,which can form hydrogen bonds with Arg599 and Ser605 in the XO domain,regu-late its affinity with the substrate xanthine,activate XO and increase the uric acid level.

Objective:To improve the differential diagnosis of systemic sclerosis (SSc) with hypertension and renal insufficiency.Methods:The clinical characteristics, diagnosis and treatment of a SSc patient with hypertension and renal insufficiency were reported and discussed.Results:A middle-aged female patient with a history of SSc for 5 years, headache and gross hematuria for 10 days was admitted. Abrupt increase in blood pressure and creatinine, glomerular hematuria, proteinuria, low complement C3 and C4, positive antinuclear antibody (ANA), anti-dsDNA antibody and perinuclear antineutrophil cytoplasmic antibody (pANCA) were presented. Renal pathology showed lupus nephritis (LN) (type Ⅳ). After glucocorticoid pulse therapy, followed by cyclophosphamide, belimumab, and symptomatic treatment, the symptoms were relievedand lupus disease activity were decreased.Conclusion:For SSc patients with increased blood pressure and creatinine, the presence of other diseases should be considered in addition to scleroderma renal crisis. Renal biopsy and pathological examination should be performed to confirm the diagnosis and avoid misdiagnosis.

Objective:To develop an online interactive cytopathological training program, and to evaluate it for improving the cytopathological diagnostic ability of endoscopists in endoscopic ultrasound-guided fine-needle aspiration (EUS-FNA) of pancreas.Methods:A total of 5 500 cytopathological images were collected from 194 patients with pancreatic solid mass who underwent EUS-FNA in Nanjing Drum Tower Hospital from August 2018 to August 2019. The cell type in each cytopathological picture was labeled by senior cellular pathologists, which was used to build a learning and testing platform for online interactive cytopathological training. Five endoscopists without cytopathological background were invited to participate in this training. Sensitivity, specificity, positive predictive value and negative predictive value of endoscopists in differential diagnosis of cancer and non-cancer before and after training were compared to evaluate the effect of the online interactive cytopathological training program on improving the ability of endoscopists in diagnosis of cytopathology.Results:A cytopathological training platform for endoscopists to learn and take online test was successfully built. Before training, sensitivity, specificity, positive predictive value, negative predictive value and accuracy of diagnosis of cancer and non-cancer for endoscopists were 0.55 (95% CI: 0.53-0.58), 0.32 (95% CI: 0.30-0.35), 0.43 (95% CI: 0.41-0.45), 0.44 (95% CI: 0.41-0.47) and 0.43 (95% CI: 0.42-0.45), respectively. After training, the above indicators were 0.96 (95% CI: 0.95-0.97), 0.70 (95% CI: 0.68-0.73), 0.74 (95% CI: 0.72-0.76), 0.95 (95% CI: 0.94-0.96) and 0.81 (95% CI: 0.80-0.83), respectively, which were significantly improved compared with those before ( P<0.001). Conclusion:The online interactive cytopathological training program can improve the understanding and diagnostic ability of endoscopists in pancreatic cytopathology, help to implement rapid on-site evaluation in the process of EUS-FNA, and improve the diagnostic efficiency of EUS-FNA.