Objective To investigate the expression of Toll-like receptor 2 and 4 (TLR2 and TLR4) in peripheral blood mononuclear cells (PBMCs) and the serum inflammatory cytokine levels in patients with liver failure. Methods The expressions of TLR2 mRNA and TLR4 mRNA in PBMCs were detected by RT-PCR in 20 healthy controls, 20 chronic hepatitis B (CHB) patients, 18 liver failure patients in early stage and 14 in intermediate-end stage. The serum contents of endotoxin, TNF-α and IL-6 were detected by ELISA. Results Compared with the healthy controls, the expression of TLR2 mRNA and TLR4 mRNA, and the contents of endotoxin, TNF-α and IL-6 increased in CHB patients and liver failure patients ( both early stage and intermediate-end stage) ( F = 32.997, 37.476, 23. 951,57. 265 and 38. 403, P < 0.01 ). TLR2 mRNA expression in liver failure patients in intermediate-end stage was higher than that in the early stage, but that for TLR4 mRNA was lower than that in early stage. The expressions of TLR2 mRNA and TLR4 mRNA in PBMCs were significantly correlated with the contents of TNF-α and IL-6 ( r = 0. 917 and 0. 788, P < 0. 01 ). Conclusion The inflammation reaction mediated by TLR2 and TLR4 might participate in the pathogenesis of liver failure.

OBJECTIVETo discuss arthroscopic technique of single Kirschner wire and suture avoiding epiphyseal line fixation for tibial intercondylar eminence fracture and its clinical results.

METHODSFrom May 2008 to December 2012, 21 patients (13 males, 8 females, ranging in age from 6 to 14 years old) with tibial intercondylar eminence fracture were treated arthroscopically with single Kirschner wire and suture avoiding epiphyseal line fixation technique. According to Meyers and McKeever classification, 7 patients were type II, 10 patients were type III, and 4 patients were type IV. Active rehabilitation began at one week after operation. The patients were followed up for 10 to 30 months. X-ray films were taken to evaluate fracture healing at 1, 3, 6 months after operation; range of motion, the anterior drawer test, the lachman test and the Lysholm knee score were used to evaluate clinical effects.

RESULTSAll fractures were healed without displacement at 6 weeks after operation. Anterior drawer test and the lachman test were both negative in all patients at 3 months after operation. Lysholm knee score was 95.5 ±2.5 at 6 months after operation, and postoperative X-ray film did not find epiphyseal line broadening or narrowing.

CONCLUSIONArthroscopic treatment for tibial eminence intercondylar fracture with single Kirschner wire and 8-shaped suture avoiding epiphyseal line fixation technique has many advantages, such as firm fixation,early mobilization, less invasive, less injury of physis and satisfactory effect.

Adolescent ; Arthroscopy ; Bone Wires ; Child ; Female ; Fracture Fixation, Internal ; Humans ; Male ; Sutures ; Tibial Fractures ; surgery

AIM:To observe the effect of Shexiang Baoxin Pills on pathologic changes and heart function of streptozotocin(STZ)-induced diabetic rats. METHODS:The rats were randomly divided into the control group,Shexiang Baoxin Pill group and STZ group.The rats in Shexiang Baoxin Pill group took Shexiang Baoxin Pill(50 mg/kg) to perfuse the stomach every day.The cardiac functions in 8 weeksafter treatment were determined.Meantime,the cardiac pathologic changes were observed through electron microscope and immunohistochemistry. RESULTS: Shexiang Baoxin Pills could reduce the interstitial fibrosis of myocardium in STZ rats.it could obviously improve the cardiac contraction and diastolic functions in STZ rats. CONCLUSION: Shexiang Baoxin Pills can(amendment) cardiac pathologic changes and improve cardiac function in STZ rats.

Objective To investigate the clinical and immune pathological features of Ullrich congenital muscular dystrophy (UCMD) with sarcolemma-specific collagen Ⅵ deficiency (SSCD). Methods The clinical aspects of 2 patients with SSCD were analyzed and the muscle specimens from them were studied by immunofluorescence. Results SSCD patients were clinically characterized by neonatal hypotonia with proximal contractures and distal hyperlaxity at birth or early infancy. Immunofluorescence staining revealed partial deficiency of collagen Ⅵ. Double immunofluorescence staining revealed sarcolemma-specific deficiency of collagen Ⅵ, while collagen Ⅳ intact in thesarcolemma. Conclusions The clinical picture and severity of UCMD with SSCD are similar to the cases with collagen Ⅵ complete deficiency. The proximal contractures and distal hyperlaxity are the clinical hallmarks of both types. Sarcolemma-specific collagen Ⅵ deficiency can be better demonstrated by double immunofluorescence staining.

Objective To evaluate the therapeutic effects of somatostatin(stilamin)and rhubarb for severe acute pancreatitis(SAP).Method A total of 42 patients with SAP received traditional treatment in combination with somatostatin(stilamin)and rhubarb,and compared with 40 SAP patients with routine treatment.The changes of acute physiology and chronic health evaluationⅡ(APACHEⅡ),serum amylas,serum creatinine,blood calcium,blood glucose,white blood cell count,the duration of abdominal pain,abdominal bulge,fast and hospital stay,complications,morlality and operation rate on the fist day,third day and fifth day were compared between two groups.Results Somatostatin and rhubarb reduced the complications,operation rate and mortality, and shortened the duration of abdominal pain,abdominal bulge,fast and hospital stay.Conclusions Combination of somatostatin and rhubarb is effective in the treatment of SAP patients.

C8orf32 is a gene which has not been functionally characterized,the mRNA level of this gene is significantly higher in breast cancer tissues than that in normal breast tissues.The amplified cDNA fragment was inserted into the pGEX-6P1 vector fused with the upstream GST gene.The expression vector was transformed into the E.coli BL21(DE3) strain and expression of GST-C8orf32 fusion protein was induced by IPTG..After removal of GST tag by site-specific protease,the C8orf32 protein fused with an eight amino acid peptide tag was obtained.The purity of recombinant C8orf32 protein was about 95%.The identity of the purified protein was confirmed by N-terminal sequencing and tandem mass spectrometry.The polyclonal antibody was prepared by immunizing the New Zealand white rabbits with C8orf32 protein.The polyclonal antibody was proved to recognize the C8orf32 protein correctly.The purified C8orf32 protein can be used for structural and functional studies and the polyclonal antibody can be used for tissue specific protein expression profiling.

ObjectiveTo investigate the application of rapid immunohistochemical staining technique in intraoperative frozen section diagnosis of thyroid neoplasm.Methods MaxVision one-step rapid immunohistochemical staining technique was used to detect the expression of CK19,HBME-1,and Gal-3 in frozen section of papillary thyroid carcinoma(PTC)andthyroid benign lesions.MaxVision conventional immunohistochemistry of frozen remaining tissue was served as control.ResultsMaxVision one-step rapid immunohistochemical staining technique could be completed in 20 minutes.The positive localizations of three markers detected by rapid immunohistochemistry were similar to conventional immunohistochemistry, in general.The expression of CK19 was located in cytoplasm and cellular membrane.Gal-3 and HBME-1 were mainly detected in follicular luminal border and/or surface of papilla. The staining intensity in rapid immunohistochemistry was stronger than that in conventional immunohistochemistry. The positive rates of CK19,HBME-1,and Gal-3 by rapid immunohistochemistry in frozen sections were: 0 (0/28),10.7 ％ (3/28),0 (0/28),respectively,for benign lesions (nodular goiter,Hashimoto thyroiditis,thyroid adenoma); and 94.9 ％(37/39),92.3 ％ (36/39),92.3 ％ (36/39),respectively,for PTC.The expression of three markers between thyroid benign lesions and PTC had a significant difference (x2 =59.326,55.861,44.605,all P ＜ 0.001).In benign lesions,the rate of same case with two and more positive markers was 0,while in PTC it was 100 ％ and significantly different (x2 =67.000,P ＜ 0.05).ConclusionMaxVision one-step rapid immunohistochemical staining technique could be applied in intraoperative frozen section diagnosis.Detecting CK19,HBME-1,and Gal-3 expression in intraoperative frozen section has an auxiliary value for diagnosis of PTC.

Objective To investigate predisposing factors for early infection in patients with severe acute pancreatitis.Methods The clinical and laboratory data including age、gender、APACHE Ⅱscore on admission、hemodiastase、mechanical ventilation、blood calcium、mean arterial blood pressure、blood glucose、 alanine aminotransferase、aspartate aminotransferase、total bilirubin、necrosis of the pancreas、hypoxemia、 entero-functional disturbance、etiological factor、serum albumin、serum creatinine、urea nitrogen and haematocrit were analyzed by multiple linear regression in relation with the infection incidence in the 86 SAP patients hospitalized from Jan 2002 to Mar 2007.Results The fasting time、hiliary panereatitis、 hypoxemia、necrosis of pancreas、entero-functional disturbance、serum creatinine、urea nitrogen and haematocrit were positively correlated with the incidence of pancreatic infection(all P

Objective:To investigate the quantification of CD4~+CD25~+ regulatory T cells and distribution of CD4~+CD8~+ T lymphocyte subgroup in peripheral blood of patients in chronic hepatitis B (CHB),and to reveal relationship between CD4~+CD25~+ regulatory T cells,CD4~+CD8~+ T lymphocyte subgroup and HBV infetion as well.Methods:CD4~+CD25~(high),CD4~+CD25~+Foxp3~+Treg and CD3~+CD4~+CD8~+T lymphocyte subgroup in peripheral blood from 50 patients with CHB and 20 healthy controls was analyzed using flow cytometry.HBV DNA was detected by fluorescence quantitative PCR.Results:The number of CD4~+CD25~(high)Tregs in patients with CHB was obviously higher than that in healthy controls(P＜0.01)and increased with copies of HBV DNA.The same with the change of CD4~+CD25~+Foxp3~+Tregs in patients with CHB and there was a positive correlation between CD4~+CD25~(high)Tregs and CD4~+CD25~+Foxp3~+Tregs(r=0.890,P＜0.001).Compared with healthy controls,the frequency of CD4~+T cells and the ratio of CD4~+/CD8~+ in patients with CHB was declined,but there was no significant difference in the frequency of CD3~+T cells and CD8~+T cells between them(P＞0.05).The variation in the number of CD4~+CD25~(high)Tregs was correlated positively with the copies of HBV DNA(r=0.782,P＜0.001)and glutamic-pyruvic transaminase(ALT)(r=0.432,P＜0.005)separately,but negatively with the frequency of CD3~+,CD4~+,CD8~+T cells and the ratio of CD4~+/CD8~+(P＞0.05).The variation in the frequency of CD3~+,CD4~+,CD8~+T cells and the ratio of CD4~+/CD8~+ was also correlated negatively with the copies of HBV DNA(P＞0.05).Conclusion:The number of CD4~+CD25~(high)Tregs increases in patients with CHB and is in accordance with the copies of HBV DNA and increased level of ALT.Further studies should be done to investigate weather CD4~+CD8~+ T lymphocyte subgroup could be used to monitor the state of community.