Effect of mesalazine on Th1, Th17 and Treg cells in mice with DSS-in-duced ulcerative colitis / 中国病理生理杂志

AIM:To investigate the effect of mesalazine treatment on regulation of Th1, Th17 and Treg cells in mice with dextran sulfate sodium ( DSS)-induced ulcerative colitis ( UC) .METHODS:The expression of IL-17, IFN-gam-ma and Foxp3 in the peripheral blood mononuclear cells ( PBMC) and intestinal mucosa lamina propria mononuclear cells ( LPMC) of DSS-induced UC mice was detected by flow cytometry analysis.The effect of mesalazine treatment on regulaiton of Th1, Th17 and Treg cells in the mice with DSS-induced ulcerative colitis was examined.RESULTS:The expression of IL-17, IFN-γand Foxp3 on CD4 +T cells were significantly higher in the PBMC of DSS-induced mice than those in control group.CD4+IFN-γ+T cells and CD4 +Foxp3 +T cells were higher in LPMC than those in control group, except CD4+IL-17 +T cells.Moreover, the Th1, Th17 and Treg cells were higher in DSS group than those in control group in LPMC.How-ever, only Tregs was higher in PBMC.Pre-treatment with mesalazine significantly decreased the number of Th17, Th1 and Treg cells of UC model mice both in PBMC and LPMC.CONCLUSION: The Th1, Th17 and Tregs cells in DSS-induced mice were significantly higher than those in control mice, suggesting that CD4 +T cell subsets play an important role in the pathogenesis of UC.Mesalazine may play a role in the treatment of UC by regulating the Th1, Th17 and Tregs cells.

P75NTR protein promotes L02 cells proliferation in vitro via TrkANGFR/P75NTR heterodimer signal pathway / 中华消化杂志

ObjectiveTo observe the expression of nerve growth factor(NGF)and its receptors (TrkANGFRand P75NTR) in hepatocytes and to explore the biological effects of exogenous P75NTR protein on hepatocytes.MethodsL02 hepatocytes were cultured in vitro.The expression of NGF,TrkANNGFR and P75NTR in L02 cells were assessed by immunocytochemistry and fluorescent quantitation polymerase chain reaction (PCR). The effects on L02 cell proliferation by exogenous P75NTR,NGF,NGF+ P75NTR,anti-TrkANGFR and anti-P75NTR were detected by XTT assay.The effect of exogenous P75NTR on L02 cell apoptosis was measured by flow cytometry (Annexin V/PI) and the effect of exogenous P75NTR on L02 cell cycle was determined by flow cytometry (PI).ResultsL02 cell proliferation was promoted by P75NTR and in dose-dependent manner.The A value of NGF group and NGF+ P75NTR group was 0.4916±0.0565 and 0.5839 ± 0.0733,respectively,and there was statistical significance compared with control group (0.3601 ± 0.0310,P＜0.05).The A value ot anti-TrkANGFR group was 0.2689±0.0229,and there was statistical significance compared with control group (P=0.003).The A value of anti P75NTR was 0.3524 ± 0.0312,and there was no statistical significance compared with control group (P=1.000). Exogenous P75NTR had anti-apoptosis effect on L02 cells,the antiapoptosis effect was strongest when 100 ng/ml P75NTR was added and the expression quantity was 3.70 ±0.26.However there was no statistical significant compared with control group (4.10 ± 0.62,P=1.000).P75NTR affected the cell cycle S phase of L02 cells and in dose-dependent manner,which was inverted U shaped curve.The effect was strongest when the concentration was 100 ng/ml (25.60 ±0.40) and there was statistical significance compared with control group (20.10 ±1.00,P=0.000).Exogenous NGF,P75NTR and NGF + P75NTR up regulated the gene expression of NGFmRNA,TrkANGFRmRNA and P75NTR mRNA in L02 cells and there was statistical significance compared with control group (P＜0.05).There was no significant difference in the gene expression of NGFmRNA,TrkANGFFR mRNA and P75NTR mRNA between anti-TrkANGFR,anti-P75NTR groUp and control group (P＞0.05).Conclusion NGF and its receptors TrkANGFR and P75NTR were expressed in L02 cells.The appropriate dose of exogenous P75NTP protein promoted L02 cells proliferation via TrkANGFR/P75NTR heterodimer signal pathway.

Study on the mechanism of apoptosis of hepatic stellate cells induced by nerve growth factor / 中国医师杂志

ObjectiveTo investigate the effect of NGF on apoptosis of HSC in vitro and explore the possible mechanism.MethodsHSC was incubated with different concentrations of NGF.HSC apoptosis was identified by FCM.The expressions of apoptosis-regulating proteins Caspase-3,p53 and Bcl-2 of HSC after apoptosis induced by NGF were examined by immunohistochemical staining.Expressions of NGF and p75NTR were detected by immunofluorescence.ResultsApoptosis index of HSC was higher than that of control group [(22.36±9.51)％ vs (5.88±1.36)％] after treated with NGF (100 ng/ml) (P＜0.05).After incubating with 100 ng/ml NGF for 24 h,the positive expression rates of p53 and Caspase-3 of HSC increased significantly than those of control group [(78.41±4.00)％ vs (34.96±3.84)％,(39.26±1.57)％ vs (9.27±1.01)％,P ＜0.05].The positive expression rate of Bcl-2 protein of HSC significantly decreased compared with that of control group (18.12±1.38)％ vs (91.53±2.98)％ (P＜0.05).When HSC was stimulated with 100 ng/ml NGF for 24 h,the average optical density of NGF increased significantly than control group (6.53±1.40 vs 1.77±0.17) (P＜0.05),while the expression of p75NTR was not significantly changed (3.52±0.36 vs 4.24±0.38) (P＞0.05).ConclusionsThe mechanism of NGF to induce HSC apoptosis may be associated with the up-regulating expression of Caspase3,P53 and down-regulating expression of Bcl-2 on HSC.NGF could be used as an initiating factor and effect factor to increase the expression of NGF on HSC,but it had no significant effect on p75NTR expression.

The preliminary clinical observation of Huganjiexian decoction in the treatment of hepatic cirrhosis / 国际中医中药杂志

Objective To observe the clinical therapeutic effects and evaluate the security of Huganjiexian decoction combined with conventional therapy on hepatic cirrhosis.MethodsBy the randomized and prospective study method,34 patients with liver cirrhosis were divided into experimental group and control group.The experimental group was treated with Huganjiexian decoction combined with conventional therapy while the control group was treated with conventional therapy alone.Patients in both groups were treated six months.At the beginning and 6 months after treatment,levels of alanine transaminase (ALT),aspartate transaminase (AST),albumin (ALB),albumin/globulin (A/G),total bilirubin (TBiL),blood urea nitrogen (BUN),serum creatinine (Scr) were determined.Results Levels ofALT、AST、TBiL decreased in both groups after being treated for six months,and the differences of downward trend of the experimental group were more significant than control group (F=36.63,40.31,38.65,P＜0.05).Levels ofALT、AST、TBiL of the experimental group were lower than those of control group significantly (F=8.67,7.62,4.36,P＜0.05 ).The A/G raised in both groups after treatment,and the upward trend of the experimental group was greatly different from that of control group (F=24.10,P＜0.05),the value of A/G of the experimental group was higher than that of control group (F=4.78,P＜0.05).The ALB raised in both groups after treatment,while the upward trend of the experimental group was no different from that of control group (F=0.89,P＞ 0.05).Thevalue of ALB had no significant changes in both groups (F=3.15,P＞0.05).Conclusion Huganjiexian decoction possessed therapeutic effect on hepatic cirrhosis,it had no obvious toxicity and side

Therapeutic effect of HuGanJieXian decoction on rats hepatic fibrosis / 国际中医中药杂志

Objective To observe therapeutic effects of HuGanJieXian decoction on rats hepatic fibrosis induced by tetrachloride. Methods Rat models of hepatic fibrosis were constructed by intraperitoneal injection of tetrachloride.HuGanJieXian decoction composed of low, middle, and high dose curcumin were given to these rats respectively at the same time. Sho-saiko-to compound treatment group and Fufangbiejiarangan Tablets treatment group were made as positive control groups. After twelve weeks, all rats were executed. Serum samples were kept for measuring serum levels of PC-Ⅲ, LN, and HA. Left livers were extirpated for pathologic examination including H.E and Masson stainings. Grade of hepatic fibrosis were evaluated according to SSS system. The levels of superoxide dismutase (SOD) and malondialdehyde (MDA) of supematant centrifugated from hepatic tissue homogenate were detected. Results Serum levels of PC-Ⅲ, LN, and HA were depressed obviously in decoction groups compared with those of fibrotic group (P<0.05) , especially in the low-dose curcumin group.HuGanJieXian Decoction could increase the level of SOD and decrease the level of MDA (P<0.05) , especially in the low-dose curcumin group. Staining of H. E and Masson showed that degrees of hepatic fibrosis in decoction groups were improved obviously compared with that of the fibrotic group. Conclusion HuGanJieXian Decoction can improve rat hepatic fibrosis, the mechanism of this effect may be associated with protecting hepatic cell membrane and anti- peroxidative damage.

Helicobacter pylori negative peptic ulcer and ulcer bleeding:a multi-center case-control study in China mainland / 中华消化内镜杂志

Objective The primary aim of this study was to examine the proportion and natural history of Helicobacter pylori (Hp) negative bleeding peptic ulcers. Methods The study was designed as a multiple-center, case-control study conducted in 14 endoscopy centers in China from April 2006 to March 2007. Each center was expected to recruit 30 peptic ulcer patients with bleeding ( PUB group) and 30 without (PU group). All screened patients with upper gastrointestinal bleeding received endoscopy within 24 hours of admission. Biopsy specimens were taken from the antrum to determine Hp infection by rapid urease test and pathology. Patients with negative Hp infection at first examination were asked to receive urease breathe test (UBT) one month later. Results A total of 617 patients were enrolled with 263 in PUB group and 354 in PU group. There is no significant difference in demographic characters between 2 groups ( P ＞0. 05). The rate of Hp infection in PUB group ( 161/263, 61.2% ) was significantly lower than that in PUgroup (311/354, 87. 9%, P ＜0. 001 ). The incidence of complex ulcer in Hp positive PUB patients was 7.5% ( 12/161 ), which is significantly higher than that in Hp negative PUB patients ( 1/102, 1.0% , P =0. 018). In PUB group, no significant differences were found between Hp positive and negative patients in regarding of age, sex, rates of haematemesis, duodenal ulcer and gastric ulcer, and size of ulcer ( P ＞0. 05 ). Among 102 Hp negative cases in PUB, no positive case was found in UBT one month later. Conclusion We have demonstrated a rise in the incidence of Hp negative bleeding ulcers in China. The idiopathic ulcer was not rare, and might have a higher tendency to cause bleed.

Observation on the therapeutic Effects of Hepatic Fibrosis of Rats with Salvia Miltiorrhiza / 国际中医中药杂志

Objective To investigate therapeutic effects of Salvia miltiorrhiza on hepatic fibrosis of rats induced by carbon tewachloridean. Methods Rat models of hepatic fibrosis were founded by intraperitoneal injection of carbon tetrachloride. Salvia miltiorrhiza were given to these rats. Normal group and control group were set for comparison at the same time. Serum levels of ALT, AST, HA, LN, and PC-Ⅲ were detected; HE and Masson staining were conducted in hepatic tissues to observe pathological variations. Results Salvia miltiorrhiza could decrease serum levels of ALT, AST, HA, LN,PC-Ⅲ obviously (P ＜0.01), compared with the control group; Salvia miltiorrhiza could obviously improve pathological variations compared with the control group. Conclusion Salvia miltiorrhiza has therapeutic effect on hepatic fibrosis of rats

Relationship between alterations of p16(INK4a) and p14(ARF) genes of CDKN2A locus and gastric carcinogenesis / 中华医学杂志(英文版)

<p><b>OBJECTIVE</b>To investigate the relationship between alterations of p16(INK4a) and p14(ARF) genes and gastric carcinogenesis.</p><p><b>METHODS</b>The tumors and neighboring gastric tissues from 48 patients with gastric cancer were studied. The homozygous deletion, mutation, methylation of the CpG islands, and mRNA expression of p16(INK4a) and p14(ARF) genes were assessed by PCR, PCR-SSCP, PCR based methylation assay, and RT-PCR.</p><p><b>RESULTS</b>(1) The homozygous deletion rate of p16(INK4a) and p14(ARF) was 35.4% (17/48), and no homozygous deletion was examined in any gastric tissue neighboring the tumor. (2) There was no point mutation of p16(INK4a) and p14(ARF) in 31 gastric cancers without homozygous deletion or in the matched gastric tissues adjacent to the tumor. (3) Methylation of the CpG islands of p16(INK4a) and p14(ARF) was detected in 47.9% (23/48) of gastric cancers, while methylation was observed only in 2 of 48 gastric tissues neighboring the cancer with a significant difference (P < 0.01). (4) The loss rate of p16(INK4a) mRNA was 47.9% (23/48) in gastric cancer, and the patients of the combined methylation of exons 1alpha and 2 had a higher loss rate (100%, 6/6) of p16(INK4a) mRNA than those of the methylation of the other exons (11.8%, 2/17, P < 0.01); the loss rate of p14(ARF) mRNA was 45.8% (22/48) in gastric cancer, and patients with the combined methylation of exons 1beta and 2 had a higher loss rate (100%, 3/3) of p14(ARF) mRNA than those of the methylation of the other exons (15%, 3/20, P < 0.05). (5) The combined loss of p16(INK4a) and p14(ARF) mRNAs was examined in 1 (5.6%) of 18 patients of well and moderately-differentiated carcinomas, and 11 (36.7%) of 30 patients of poorly and not-differentiated carcinomas with a significant difference (P < 0.05).</p><p><b>CONCLUSION</b>p16(INK4a) and p14(ARF) genes are frequently inactivated by homozygous deletion and methylation of the 5'CpG islands in gastric cancer, which may play an important role in the carcinogenesis of gastric cancer.</p>

The blind area in routine gastric lavage and its clinical significance / 中华消化内镜杂志

Objective To observe the blind area in routine gastric lavage and evaluate its significance. Methods Each of 10 healthy volunteers drank 60 ml water containing 185 MBq 99mTc. Images of mouth, esophagus and stomach of all subjects were collected from anterior position with gamma camera, and intraoral and intraesophageal radiocounts were monitored with " Region of Interesting" technique at 0, 0.5, 1,2,3 and 4 hours. Ratios of the counts at subsequent time points vs the count of 0 hour were calculated. For other 13 volunteers, 100 ml, 200 ml, 300 ml and 400 ml normal saline containing 25% compound meglucamine diatrizoate were drunk and injected into stomachs through stomach tube respectively. After that, X-ray chest films were taken at the dorsal and left lateral decubitus positions to observe the oral and e-sophageal developments. Results The images of mouths and esophagus of 10 subjects could be identified more than 4 hours after the administration of 99mTc. The ratios of oral counts and esophageal counts at 0. 5 ,1, 2, 3 and 4 hrs were 30. 7% , 10. 5% , 6. 3% , 4. 7% , 3. 7% , and 18. 6% , 4. 4% , 3. 5% , 3. 0% ,2. 7% respectively. The ratios of both oral and esophageal counts were 49. 3% , 14. 9% , 9. 8% , 7. 7% and 6.4%. When the liquid was drunk, full oral and esophageal developments could be identified. While the liquid was injected through the tube, no development was found. Conclusions Liquid drug may remain in mouth and esophagus for a long period of time after administration. Routine gastric lavage through nasogastric tube can not clean the oral and esophageal drug residues. There are oral and esophageal blind areas when routine gastric lavage is employed.

Cloning of P1 and P3 promoters of human insulin-like growth factorⅡ gene / 中国病理生理杂志

AIM: To clone P1 and P3 promoters of the human insulin-like growth factor Ⅱ(IGF-Ⅱ) gene. METHODS: According to the complete DNA sequence of IGF-Ⅱ gene, the nested primer PCR was performed for amplifying P1 and P3 promoter fragments of the gene from human L-02 cell line.These PCR products were analyzed by agarose gel electrophoresis, and cloned by using TOPO TA Cloning kit. The positive clones containing P1 and P3 fragments were selected and confirmed by sequencing.RESULTS: The DNA sequences of P1 and P3 promoters cloned were accordant with GenBank data. CONCLUSION: In this study P1 and P3 promoters of the IGF-II gene were cloned successfully.