[Effect of Hanfangji compound on proliferation of hepatic stellate cells and expressions of TTR, ITIH1 and SERPINF2].

OBJECTIVE: To observe and compare the effects of Hanfangji Compound and IFN-gamma on expressions of transthyretin (TTR) , inter-alpha inhibitor H1 (ITIH1) and serpin peptidase inhibitor clade F member 2 (SERPINF2) of hepatic stellate cells (HSC-T6). METHOD: Hanfangji Compound and IFN-gammaof different concentrations were used in hepatic stellate cell-T6 (HSC-T6) for 48 h. Flow cytometer was used to detect the effects of Hanfangji Compound and IFN-gamma on HSC proliferation. RT-PCR method was adopted to detect mRNA expressions of TFR, ITIH1 and SERPINF2. TTR, ITIH1 and SERPINF2 secretions were detected by ELISA. The protein localizations of TTR, ITIH1 and SERPINF2 were examined by immune fluorescence. The protein expression of TfR and ITIHI were determined by Western blot. RESULT: After Hanfangji Compound and IFN-gamma were adopted in HSC-T6, compared with the control group, the cell proliferation was inhibited obviously (P < 0. 05) , protein expressions of TTR, ITIH1 and SERPINF2 and mRNA expression increased significantly, with certain correlation with concentrations of Hanfangji Compound. The 2. 5 g L-I Hanfangji Compound group was superior to the IFN-gamma group (P <0. 05). CONCLUSION: Hanfangji Compound can inhibit HSC proliferation, upregulated TTR, ITIH1 and SERPINF2 proteins and mRNA expression, which may be one of mechanisms of anti-hepatic fibrosis of Hanfangji Compound.

Effect of Zhizhu Kuanzhong Capsules () on Treatment of Functional Dyspepsia: A Meta-Analysis of Randomized Controlled Trials.

OBJECTIVE: To evaluate the effect of Zhizhu Kuanzhong Capsules (, ZKC) for functional dyspepsia (FD) through meta-analysis. METHODS: Online databases, including PubMed, EM base, China National Knowledge Infrastructure, Wanfang Data, VIP database and Cochrane Library, were searched for randomized controlled trials (RCTs) of ZKC for FD from the inception to April, 2016. Trials were selected according to inclusion criteria and were evaluated with quality assessment standards in the Cochrane Handbook for Systematic Reviews of Interventions and Jadad scale. RevMan 5.3 and GRADEprofiler 3.6 were used for statistical analysis and evidence quality assessment. RESULTS: Twenty-three trials with 2,496 patients were included and most of them were of poor methodological quality. ZKC alone or ZKC combined with routine Western medicine (WM) showed a better clinical effect rate compared with the control group of WM [odds ratio (OR)=3.32, 95% confidence interval (2.66, 4.15), P<0.00001]. No serious adverse reactions were reported. CONCLUSIONS: ZKC alone or ZKC combined with routine WM could significantly improve the clinical effective rate in the treatment of FD. The quality of the evidence is low, so it is necessary to design multicenter, strictly randomized and double-blind controlled trials with large samples to validate the conclusions.

[Role of changes in sodium pump activity and endoplasmic reticulum stress in the ischemia/reperfusion induced injury of isolated hearts].

OBJECTIVE: To investigate the roles of change in sodium pump activity and endoplasmic reticulum stress (ERS) in ischemia/reperfusion (I/R) injury of the isolated rat hearts. METHODS: Sixty male SD rats were randomly divided into 6 groups (n=10 each):normal control group (NC), I/R group (I/R), ouabain-I/R group (OUA-I/R), anti-digoxin antiserum-I/R group (Anti-Dig-I/R), PP2 (Src kinase inhibitor)-ouabain-I/R group (PP2-OUA-I/R),U73122 (PLC inhibitor)-ouabain-I/R group (PP2-OUA-I/R). The isolated rat hearts were perfused on the Langendorff apparatus. Except for NC group, all the hearts were subjected to 30 min global ischemia and followed by 120 min reperfusion. The cardiac function indexes were recorded at the same time. The coronary effluent was collected for estimating the activity of lactate dehydrogenase (LDH) and creatine kinase (CK). The activity of Na+-K+-ATPase and intracellular calcium concentration in myocardial tissue were measured. Apoptosis was evaluated by Flow cytometric analysis. The expressions of sodium pump α1 subunit, glucose regulated protein(GRP78),C/EBP homologous protein (CHOP) and Bcl-2/Bax were determined by Western blot. RESULTS: Pretreatment with ouabain significantly reduced the recovery of cardiac function, increased the levels of CK, LDH and intracellular calcium concentration, decreased the activity of Na+-K+-ATPase. In addition, ouabain markedly increased the myocardial apoptosis index, down-regulated the expressions of sodium pump α1 subunit and Bcl-2, up-regulated the expressions of GRP78,CHOP and Bax; while these changes were significantly improved in the Anti-Dig-I/R group compared with those in the I/R group; PP2 or U73122 partially blocked the effects of ouabain on myocardial I/R injury. Compared with the OUA-IR group, the recovery of cardiac function, the activity of Na+-K+-ATPase and the expressions of sodium pump α1 subunit and Bcl-2 were significant higher, meanwhile the leakage of CK and LDH, intracellular calcium concentration, myocardial apoptosis index and the expressions of GRP78 and Bax were significantly lower in PP2-OUA-I/R and U73122-OUA-I/R group. CONCLUSIONS: Changes in sodium pump function and endoplasmic reticulum stress all participate in the process of I/R injury. Current findings further suggest that sodium pump mediates ERS by activating signals of Src and PLC pathway, which may be one of the mechanisms of apoptosis induced by I/R injury.

The Efficacy of Aspirin in Preventing the Recurrence of Colorectal Adenoma: a Renewed Meta-Analysis of Randomized Trials.

BACKGROUND: Through search the possible randomized control trials, we make a renewed meta-analysis in order to assess the impact of aspirin in preventing the recurrence of colorectal adenoma. MATERIALS AND METHODS: The Medicine/PubMed, Embase, Cochrane Central Register of Controlled Trials (CENTRAL), Chinese biomedical literature service system (SinoMed) databases were searched for the related randomized controlled trials until to the April 2016. Three different authors respectively evaluated the quality of studies and extracted data, and we used the STATA software to analyze, investigate heterogeneity between the data, using the fixed-effects model to calculate and merge data. RESULTS: 7 papers were included the renewed meta- analysis, among these studies, two pairs were identified as representing the same study population, with the only difference being the duration of follow-up. Thus there were only five papers included our meta-analysis, and one Chinese paper were also included the work. Results were categorized by the length of follow-up, different kinds of people, varied dose of oral aspirin. The relative of adenoma in patients taking aspirin vs placebo were 0.73 (95% CI 0.55-0.98, P=0.039) with 1 year follow up; 0.84 (95% CI 0.72-0.98, P=0.484) with greater than 1 year follow up; for the advanced adenoma, the RR 0.68 (95% CI 0.49-0.94, P=0.582),for one year; RR=0.75 (95% CI 0.52-1.07, P=0.552) for greater one year. Furthermore the white population could divided into two subgroups according to the different length of follow-up time. When the length of follow-up time less than 3-year, The RR of two subgroups respective were RR=0.86 (95% CI 0.76-0.98, P=0.332), I2=0%, RR=0.68 (95% CI 0.47-0.98, P=0.552), I2=64.6%, But with the extension of follow-up time greater than 2-year, with the white, oral aspirin without considering dose had no efficacy on preventing the recurrence of any adenoma, the RR was 0.86 (95% CI 0.71-1.05, P=0.302), I2=16.4%. CONCLUSIONS: This meta-analysis indicated that oral aspirin is associated with a remarkable decrease in the recurrence of any adenoma and advanced adenomas in patients follow-up for 1 year without concerning the dose of aspirin, but with the extension of follow-up time for greater than 1 year, oral aspirin can be effective on preventing the recurrence of any adenoma, but for the advanced adenoma, the result indicated that oral aspirin had no efficacy, According to the inclusion of ethnic groups, we also divided relevant papers into two subgroups as the yellow and white group. Then the follow-up time was less than 3 years, oral aspirin without considering the dose, had an significant efficacy on preventing the recurrence of any adenoma. But with the follow-up greater than 2 years, oral aspirin had no effect in the white.

Systemic immune responses to oral administration of recombinant attenuated Salmonella typhimurium expressing Helicobacter pylori urease in mice.

AIM: To evaluate whether attenuated Salmonella typhimurium producing Helicobacter pylori (H pylori) urease subunit B (UreB) could induce systemic immune responses against H pylori infection. METHODS: Attenuated S. typhimurium SL3261 was used as a live carrier of plasmid pTC01-UreB, which encodes recombinant H pylori UreB protein. Balb/c mice were given oral immunization with two doses of SL3261/pTC01-UreB at a 3-wk interval. Twelve weeks after oral immunization of mice, serum IgG antibodies were evaluated by ELISA assay. Gamma interferon (IFN-gamma) and interleukin 10 (IL-10) in the supernatant of spleen cell culture were also assessed by ELISA. RESULTS: After oral immunization of mice, serum specific IgG antibodies against UreB in vaccine group were much higher than that in PBS and native Salmonella SL3261 control groups (A450, 0.373+/-0.100 vs 0.053+/-0.022, 0.142+/-0.039, respectively, P<0.01). Moreover, IFN-gamma in vaccine group was on average 167.53+/-29.93 pg/mL, which showed a significant increase vs that of PBS control group (35.68+/-3.55 pg/mL, P<0.01). There was also a tremendous increase of IL-10 in vaccine group compared to PBS and SL3261 control groups (275.13+/-27.65 pg/mL vs 56.00+/-7.15 pg/mL, 68.02+/-15.03 pg/mL, respectively, P<0.01). In addition, no obvious side effects in mice and no change in gastric inflammation were observed. CONCLUSION: The multiple oral immunizations with the attenuated S. typhimurium expressing H pylori UreB could induce significant systemic immune responses, suggesting it may be used as oral vaccine against H pylori infection.

[Development of a multiple ligator and its endoscopic therapy for patients with esophageal varices].

The ligators we have developed is a kind of economical and effective six-ring ligator. Endoscopic variceal ligation (EVL) was performed to treat bleeding from esophageal varices in patients with liver cirrhosis using self-made ligator and foreign multiple ligator. There are similar effects with both self-made ligator and foreign mutiple ligator in the control of variceal bleeding, variceal obliteration and rebleeding (93.8%, 87.5%, 0 in the group with self-made ligator, 94.5%, 87.1%, 2.4% in the group with foreign multiple ligator, P>0.05). In terms of the quality index, successful operation rate, hemastatic rate, variceal obliteration rate, rebleeding rate, complications and variceal recurrence rate, the self-made ligator is as good as the foreign multiple ligator, but much cheaper.

Helicobacter pylori and pancreatic cancer risk: a meta- analysis based on 2,049 cases and 2,861 controls.

AIM: Helicobacter pylori (H. pylori) have been considered as a risk factor for many cancers. We conducted this meta-analysis to clarify the association between H. pylori infection and the risk of pancreatic cancer. METHODS: We searched the Medicine/Pubmed and Embase databases, studies about the association between H. pylori infection and pancreatic cancer published up to Jan.2014 were included. Finally, a total of 9 studies were used for this a meta-analysis. The odds ratios (ORs) and 95% confidence interval (95%CI) of H. pylori infection on pancreatic cancer with respect to control groups were evaluated. Two authors independently assessed the methodological quality and extracted data. This meta-analysis was conducted using software, state (version 12.0) to investigate heterogeneity among individual studies and to summarize the studies. Using the fixed-effects or random-effects model, depending on the absence or presence of significant heterogeneity. Sensitivity analysis was performed to assess the influence of each individual study on the pooled ORs by omitting a single study each time. Publication bias was evaluated by funnel plot, using Egger's and Begg's tests. RESULTS: There was no significant association between H. pylori infection and pancreatic cancer risk in the summary ORs,(OR=1.06, 95%CI: 0.74-1.37) through the random-effect method, but heterogeneity among studies was significant (I2=58.9%), so we put the studies into two subgraphs (eastern and western). The results about western (OR=1.14 95%CI:0.89, 1.40) showed heterogeneity among the western countries of I2=6.6%, with no significant association between Hp+ and pancreatic cancer, but the eastern countries (OR=0.62, 95%CI:0.49, 0.76), I2=0, suggested that decreasing pancreas-cancer risk in subjects with Hp+ infection. Simultaneously, 7 studies examined CagA+ strains was (OR=0.84 95%CI:0.63, 1.04), I2=36% with the random-effect method, subgraphs indicated that CagA+ could decrease the risk of pancreatic cancer in the eastern subjects (OR=0.66, 95%CI:0.52-0.80), but the association was not statistically significant in the western subjects (OR=0.95, 95%CI:0.73, 1.16). CONCLUSION: Hp+ and CagA+ infection are associated with a decreased risk of pancreatic cancer in eastern populations but have no significant associations in western countries.

ß-catenin is overexpressed in hepatic fibrosis and blockage of Wnt/ß-catenin signaling inhibits hepatic stellate cell activation.

ß-catenin, a core component of Wnt/ß-catenin signaling, has been shown to be an important regulator of cellular proliferation and differentiation. Abnormal activation of Wnt/ß-catenin signaling promotes tissue fibrogenesis. In the present study, the role of ß-catenin during liver fibrogenesis was analyzed and the functional effects of ß-catenin gene silencing in hepatic stellate cells (HSCs) using small interfering (si)RNA were investigated. The expression of ß-catenin in human hepatic fibrosis tissues of different grades and normal human hepatic tissues was examined using immunohistochemistry. To inhibit the Wnt/ß-catenin signaling pathway, siRNA for ß-catenin was developed and transiently transfected into HSC-T6 cells using Lipofectamine 2000. ß-catenin expression was evaluated by quantitative polymerase chain reaction (qPCR) and western blot analysis. The expression of collagen types Ⅰ and Ⅲ was evaluated by qPCR and immunofluorescent staining. Cellular proliferation and the cell cycle were analyzed using a methyl thiazolyl tetrazolium assay. Apoptosis was assessed by Annexin V staining. A higher expression level of ß-catenin was identified in the patients with high-grade hepatic fibrosis in comparison with that of the normal controls. Additionally, ß-catenin siRNA molecules were successfully transfected into HSCs and induced inhibition of ß-catenin expression in a time-dependent manner. ß-catenin siRNA treatment also inhibited synthesis of collagen types Ⅰ and Ⅲ in transfected HSCs. Furthermore, compared with those of the control group, siRNA-mediated knockdown of ß-catenin in HSC-T6 cells inhibited cell proliferation and resulted in cell apoptosis. This study suggests a significant functional role for ß-catenin in the development of liver fibrosis and demonstrates that downregulation of the Wnt/ß-catenin signaling pathway inhibits HSC activation. Thus, this study provides a novel strategy for the treatment of hepatic fibrosis.

[Influence of negative pressure wound therapy on the angiogenesis of wounds in diabetic rats].

OBJECTIVE: To observe the influence of negative pressure wound therapy on the angiogenesis of wounds in diabetic rats. METHODS: Diabetes model was reproduced by intraperitoneal injection of 20 g/L streptozotocin in the dosage of 65 mg/kg in 40 SD rats. Two weeks later, rats were divided into control group (C) and negative pressure group (NP) according to the random number table, with 20 rats in each group. A piece of full-thickness skin in the center of the back of each rat in the size of 2 cm×2 cm was excised to produce a wound. Immediately after injury, wounds in group C were given conventional dressing change; wounds in group NP were treated with continuous negative pressure (-16.0 kPa) therapy for four hours a day, which lasted for seven days. (1) Blood glucose and body weight of rats in two groups were respectively measured by glucose meter and electronic scale before treatment, and 1 and 2 week (s) after. (2) Wound blood flow was detected by laser Doppler perfusion imager before treatment and on post treatment day (PTD) 1, 3, 7, with 5 rats at each time point. (3) On PTD 3 and 7, respectively, five rats from each group were sacrificed. The wound tissue was excised and divided into two parts. The angiogenesis in the left part tissue was observed with immunohistochemical staining. The microvessel density was calculated. (4) The full-thickness skin excised before treatment and the right part tissue freeze on PTD 3 and 7 were collected. On PTD 1 and 14, wound tissue was excised in the above-mentioned method. The mRNA levels of the vascular endothelial growth factor (VEGF), vascular endothelial growth factor receptor 1 (Fit-1), angiopoietin 1 (Ang-1), Ang-2, and tyrosine kinase receptor 2 (Tie-2) were determined with real-time fluorescence quantification PCR. Data were processed with two-way analysis of variance or LSD-t test. RESULTS: (1) No significant difference was observed between two groups in blood glucose level and body weight as a whole or at each time point (with F values respectively 0.667, 0.176, t values from 0.311 to 0.707, P values all above 0.05). (2) The difference in the overall wound blood flow of rats between two groups was significant (F = 24.66, P < 0.05). On PTD 1, 3, 7, values of wound blood flow of rats in group NP were (179 ± 24), (219 ± 12), (192 ± 30) perfusion unit, significantly higher than those of rats in group C[(127 ± 16), (179 ± 8), (144 ± 17) perfusion unit, with t values respectively 3.71, 5.57, 2.77, P < 0.05 or P < 0.01]. (3) The difference in the overall microvessel density in the wound of rats between two groups was significant (F = 33.25, P < 0.05). On PTD 3, the microvessel density in the wound of rats in group NP was (80 ± 12) per 100-time visual field, which was significantly higher than that of group C[(38 ± 4) per 100-time visual field, t = 9.257, P < 0.05]. On PTD 7, the microvessel density in the wound of rats in two groups were close (t = 1.159, P > 0.05), but the vessels in group NP were regularly arranged with spacious lumen, while the vessels in group C were disorderly arranged with narrow lumen. (4) On PTD 1, 3, mRNA expression levels of VEGF, Fit-1, and Ang-1 in group NP were obviously higher than those in group C (with t values from 1.28 to 11.60, P values all below 0.01). On PTD 7, the mRNA expression level of Ang-1 (27.59 ± 3.55) in group NP was obviously higher than that in group C (19.87 ± 1.86, t = 7.23, P < 0.001), while the mRNA level of its antagonist Ang-2 (5.79 ± 0.61) in group NP was obviously lower than that in group C (17.62 ± 0.85, t = 19.88, P < 0.001). On PTD 3, 7, 14, mRNA levels of Tie-2 in group NP were obviously lower than those in group C (with t values from 8.92 to 15.60, P values all below 0.01). CONCLUSIONS: Negative pressure wound therapy may promote wound angiogenesis by enhancing the expression of Ang-1 and lowering the expression of Ang-2 in diabetic rats.

[Effects of adipose-derived stem cells on renal injury in burn mice with sepsis].

OBJECTIVE: To investigate the effect of adipose-derived stem cells (ADSC) on renal injury in mice with burn injury and sepsis and its underlying mechanism. METHODS: (1) Adipose tissue was collected from both inguinal regions of 5 C57BL/6J mice to isolate, culture and purify ADSC through enzyme digestion, density gradient centrifugation, and adherence method. Cells of the third passage were used in the experiment. The morphologic change in cells was observed and the growth curve of cells was determined. The expression of cell surface antigen phenotype was analyzed by flow cytometry, and the cells were identified by adipogenic and osteogenic differentiation. (2) Another 37 C57BL/6J mice were divided into normal control group (n = 5), saline group (n = 16), and group ADSC (n = 16) according to the random number table. The mice in saline group and group ADSC were injected with Pseudomonas aeruginosa after being subjected to 15% TBSA full-thickness burn on the back to reproduce septic burn model. Then the mice were injected with saline and ADSC through tail vein respectively. At post burn hour (PBH) 12, 24, 48, and 72, the pathological change in kidney tissue was observed, the levels of blood urea nitrogen and serum creatinine were determined, and the levels of TNF-α, IL-12, IL-10, and cyclooxygenase-2 (COX2) mRNA were determined with real-time fluorescence quantitative PCR in both groups. Above-mentioned indexes were also examined in the normal control group (without burn). Data were processed with multifactor analysis of variance and LSD- t test. RESULTS: (1) Cells in the third passage were orderly arranged with the shape similar to fibroblasts. The percentages of CD90(+), CD105(+), CD34(-), and CD45(-) cells were all above 90%. The cells could differentiate into osteoblasts and adipocytes. The cells were identified to be ADSC. (2) From PBH 12 to PBH 72, the neutrophil infiltration gradually increased, and the structure of kidney tubules and glomeruli were deranged in saline group. The pathological change in kidney tissue in group ADSC was less serious than that of normal control group at each time point. From PBH 12 to PBH 72, the levels of blood urea nitrogen and serum creatinine in saline group were significantly higher than those of normal control group and group ADSC (P values all below 0.01). Compared with those of the normal control group, the levels of TNF-α and IL-12 mRNA were higher in group ADSC and saline group at PBH 24 (P values all below 0.05). At PBH 24, the level of TNF-α mRNA in group ADSC (1.58 ± 0.19) was lower than that of saline group (3.36 ± 0.30, P < 0.05). At PBH 24, the levels of IL-10 and COX2 mRNA in group ADSC (2.89 ± 0.47, 4.90 ± 0.59) were higher than those in normal control group (1.00 ± 0.15, 1.00 ± 0.27) and saline group (1.32 ± 0.38, 1.57 ± 0.38, P values all below 0.05). CONCLUSIONS: ADSC can decrease the levels of blood urea nitrogen and serum creatinine, promote the production of anti-inflammatory cytokines IL-10 and COX2, and reduce the release of the pro-inflammatory cytokines TNF-α and IL-12 to offer protective effects against renal injury in burn mice with sepsis.

[Repair of extensive deep burn wounds in late stage with free muscle flap].

OBJECTIVE: To explore the means for the reconstruction of extensive deep burn wounds with exposure of bone and joint in late stage. METHODS: Among all the patients with extensive deep burn hospitalized between January 2009 and May 2011, 5 patients presented wounds with exposure of bone and joint in the late stage of treatment that could not be covered by free skin grafts or flaps. Two of the five patients had more than 2 and the other 3 patients had only one such wound(s). The wound size ranged from 8 cm×5 cm to 21 cm×8 cm. Wounds were repaired by transplantation of 7 free muscle flaps (including 4 free rectus abdominis flaps and 3 latissimus dorsi flaps) combined with split-thickness skin grafts harvested from scalp. RESULTS: All the muscle flaps and skin grafts survived. Wounds with bone and joint exposure healed well. At one-year follow-up of some patients, good appearance of repaired areas and normal function of joints were observed with no signs of ulceration, arthritis, or osteomyelitis. CONCLUSIONS: Transplantation of free muscle flaps combined with split-thickness skin grafts harvested from the scalp provides satisfactory reconstruction for wounds with deep tissue exposure in patients with a shortage of skin donor site.

[Effects of Wnt/beta-catenin signaling on the phenotype change of human dermal fibroblasts and its mechanism].

OBJECTIVE: To study the role of Wnt/beta-catenin signaling in the phenotype change of normal skin fibroblasts (NFb) into myofibroblasts and the underlying mechanism. METHODS: NFb were isolated by collagenase digestion and cultured. (1) Experiment one. NFb were divided into four groups according to the random number table. Cells in control group were cultured with serum-free DMEM nutrient solution (briefly called nutrient solution). Cells in TGF-beta1 group were cultured with nutrient solution containing 10 ng/mL recombinant human TGF-beta1 (the same concentration for following experiments). Cells in Wnt3a group were cultured with nutrient solution containing 150 ng/mL Wnt3a (the same concentration for following experiments). Cells in TGF-beta1 + Wnt3a group were cultured with nutrient solution containing TGF-beta1 and Wnt3a. The mRNA and protein expression levels of beta-catenin and alpha-smooth muscle actin (alpha-SMA) were determined by real-time fluorescent quantitative PCR and Western blotting at post culture hour (PCH) 48. (2) Experiment two. NFb were divided into four groups according to the random number table. Cells in control group and TGF-beta1 group were treated as those in the corresponding groups in experiment one. Cells in SB415286 (glycogen synthase kinase-3beta inhibitor) group were cultured with nutrient solution containing 10 micromol/L SB415286 (the same concentration for following experiments). Cells in TGF-beta1 + SB415286 group were cultured with nutrient solution containing TGF-beta1 and SB415286. The mRNA and protein expression levels of alpha-SMA were determined by real-time fluorescent quantitative PCR and Western blotting, and the alpha-SMA-positive myofibroblasts were detected by immunofluorescence cytochemical staining at PCH 48. The experiments were all repeated for three times. Data were processed with analysis of variance and LSD- t test. RESULTS: (1) Experiment one. There was no statistically significant difference among four groups in beta-catenin mRNA level (F = 0.302, P = 0.823). There were statistically significant differences among four groups in beta-catenin protein level (F = 16.713, P = 0.001). The protein level of beta-catenin was higher in TGF-beta1 group (0.73 +/- 0.12) and Wnt3a group (0.82 +/- 0.17) than in control group (0.34 +/- 0.11, with t values respectively 3.028, 3.727, P < 0.05 or P < 0.01). The protein level of beta-catenin in TGF-beta1 + Wnt3a group (1.23 +/- 0.21) was higher than that of the other three groups (with t values respectively 6.911, 3.883, 3.184, P values all below 0.01). There were statistically significant differences among four groups in alpha-SMA mRNA level (F = 31.830, P = 0.001). Compared with that of control group, the expression level of alpha-SMA mRNA was up-regulated in TGF-beta1 group and down-regulated in Wnt3a group (with t values respectively 6.759, 2.535, P < 0.05 or P < 0.01). The expression level of alpha-SMA mRNA in TGF-beta1 + Wnt3a group was lower than that of TGF-beta1 group (t = 4.532, P < 0.01). The protein levels of alpha-SMA in control, TGF-beta1, Wnt3a, and TGF-beta1 + Wnt3a groups were respectively 0.83 +/- 0.17, 1.43 +/- 0.20, 0.53 +/- 0.12, and 0.89 +/- 0.14 (F = 16.597, P = 0.001). Compared with that of control group, the protein level of alpha-SMA was up-regulated in TGF-beta1 group and down-regulated in Wnt3a group (with t values respectively 4.582, 2.291, P < 0.05 or P < 0.01). The protein level of alpha-SMA in TGF-beta1 + Wnt3a group was lower than that of TGF-beta1 group (t = 4.123, P < 0.01). (2) Experiment two. There were statistically significant differences among four groups in alpha-SMA mRNA level (F = 34.101, P = 0.001). The alpha-SMA mRNA level in SB415286 group was lower than that of control group (t = 2.511, P < 0.05). The alpha-SMA mRNA level in TGF-beta1 + SB415286 group was lower than that of TGF-beta1 group (t = 3.587, P < 0.01). There were statistically significant differences among four groups in alpha-SMA protein level (F = 11.381, P = 0.003). The alpha-SMA protein level was lower in SB415286 group than in control group (t = 2.364, P < 0.05). The alpha-SMA protein level was down-regulated in SB415286 +TGF-beta1 group as compared with that of TGF-beta1 group (t = 2.556, P < 0.05). There were few alpha-SMA-positive fibroblasts in control group. Compared with that of control group, the expression of alpha-SMA was significantly increased in TGF-beta1 group (t =11.198, P < 0.01), and the expression of alpha-SMA was down-regulated in SB415286 group. Meanwhile, the expression of alpha-SMA in TGF-beta1 + SB415286 group were significantly lower than that of TGF-beta1 group (t = 5.902, P < 0.01). CONCLUSIONS: The Wnt/beta-catenin signaling might be involved in the fibroblasts-myofibroblasts transition, and it negatively regulate the TGF-beta1 -mediated profibrotic effects.

Inhibition of high-mobility group box 1 expression by siRNA in rat hepatic stellate cells.

AIM: To explore the role of high-mobility group box 1 (HMGB1) protein during liver fibrogenesis and investigate the functional effects of HMGB1 gene silencing in hepatic stellate cells (HSCs) using siRNA. METHODS: Hepatic fibrosis in rats was induced throu-gh serial subcutaneous injections of dimethylnitrosamine, and expression of HMGB1 was detected by immunohistochemistry. HMGB1 siRNAs were developed and transiently transfected into HSC-T6 cells using Lipofectamine 2000. HMGB1 expression was evaluated by real-time polymerase chain reaction (PCR) and Western blotting analysis. Expression of α-smooth muscle actin (α-SMA) and collagen types I and III was evaluated by real-time PCR. Cell proliferation and the cell cycle were determined using the methyl thiazolyl tetrazolium method. Finally, collagen content in HSC supernatant was evaluated by an enzyme-linked immunosorbent assay. RESULTS: The results showed that HMGB1 was upregulated during liver fibrosis and that its expression was closely correlated with the deposition of collagen. siRNA molecules were successfully transfected into HSCs and induced inhibition of HMGB1 expression in a time-dependent manner. Moreover, HMGB1 siRNA treatment inhibited synthesis of α-SMA and collagen types I and III in transfected HSCs. CONCLUSION: This study suggests a significant fun-ctional role for HMGB1 in the development of liver fibrosis. It also demonstrates that downregulation of HMGB1 expression might be a potential strategy to treat liver fibrosis.

[Microsurgical repair of skin and soft tissue defects on head, face, and neck].

OBJECTIVE: To study the microsurgical method of repairing skin and soft tissue defects on head, face, and neck. METHODS: Thirty-one patients with skin and soft tissue defects on the head, face, or neck were hospitalized from July 2007 to May 2010, including 10 cases of scalp defects, 4 cases of skin and soft tissue defects on face, and 17 cases of skin and soft tissue defects on neck. Among them, the cause in 20 cases was trauma, and in 11 cases they were secondary to release of cicatricial contraction. Free flaps were transplanted to repair the wounds, including 13 latissimus dorsi flaps, 3 lateral thoracic flaps, 5 scapular flaps, and 10 anterolateral thigh flaps. The area of flaps ranged from 8 cm × 5 cm to 25 cm × 18 cm. RESULTS: All flaps survived, and all the wounds healed by first intention. The average length of hospital stay was 16.7 days. Twenty-eight patients were followed up for 2 months, and in all of them satisfactory function and appearance were restored. CONCLUSIONS: Free flap graft based on microsurgery can repair wound of skin and soft tissue defects on head, face and neck by a single operation, which eases suffering of patients, and shortens the length of hospital stay.

[Repair of devastating wounds with free composite tissue flap].

OBJECTIVE: To study the safety and effects of free composite tissue flaps in repairing devastating wounds in early stage. METHODS: One hundred and twenty-three patients with 128 devastating wounds hospitalized in our burns center from 2005 to 2009 were repaired with free flaps or composite tissue flaps. Flap types used included 58 latissimus dorsi muscular flaps, 32 anterolateral thigh flaps, 21 circumflex scapular flaps, 6 dorsalis pedis composite flaps, 3 big toe nail skin flaps, 3 forearm flaps, and 1 lateral thoracic flap. One wound was repaired with lateral lower leg flap with fibula, and 3 wounds with free latissimus dorsi muscular flap plus skin graft. RESULTS: Vascular crisis was observed in 10 transplanted flaps 1 to 5 days after operation; 6 flaps with this complication were saved after emergency surgical exploration. Total survival rate of transplanted flaps and composite tissue flaps was 95.3% (122/128). All patients were followed up for 3 months to 4 years; satisfactory appearance and restoration of partial function were found in all of them. CONCLUSIONS: Free composite tissue transplantation reduces amputation rate, achieves primary reconstruction of function with good appearance, shortens length of hospital stay, and reduces surgical operation time, complications, and treatment cost. It is a good approach in the repair of massive devastating soft tissue injury.

[Analysis of drug use of 290 burn patients].

OBJECTIVE: To analyze the drug use in our burn ward, and to investigate the methods in controlling the proportion of drug expenses, as well as the rational use of antibiotics. METHODS: A randomized sample of 290 burn-patients with hospital stay more than 5 days in 2005 were enrolled in the study, and information about their drug use, (especially that of antibiotics), the drug expenses and its proportion in the total medical expenses, the total therapeutic expenses, and the healing rate of the patients were statically analyzed. RESULTS: Sulfamido was dominant among the topically applied antibiotics, while cephalosporins was dominant among the systemic applied antibiotics. The drug expenses accounted for (11 +/- 5)% in the therapy expenses, while the antibiotics expenses accounted for (5.8 +/- 1.7)% in the therapy expenses, but it accounted for (51 +/- 17)% in the drug expenses. The average expense of every patient was 22026.09 RMB, and the healing rate of the patients was 96.90%. CONCLUSION: The proportion of drug expense (especially that of antibiotics), as well as total medical expenses of burn patients can be lowered through a combined therapy.