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

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.

Objective To increase stone-breaking efficiency by improving waterway system of extracorporeal shock wave lithotripsy. Methods A water-storing bag, which possessed warm-up function and was filled with anticorrosive and antirust no-air water, and a sealed water tank were added to water-entering pipeline of shock wave generator. Water pressure pressing water-storing bag was utilized to control no-air water to be filled into shock wave generator to minimize bubble forming in shock wave outer conduct passway. Meanwhile, a temporary water-storing tank and a single-directed valve were added between pipelines and water-storing bag. Results After improving, lithotripsy could recycle no-air water to be filled into shock wave generator. It not only increased shock wave conducting and stone breaking efficiency, but also decreasee air content in shock wave generator and outer conduct passway, and decreased bubble forming, thus reducing complication in more rational procedures. Conclusion The method of improving waterway system of extracorporeal shock wave lithotripsy is simple, cheap, effective and worth popularizing.

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 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.

OBJECTIVE: To evaluate the immuno-modulatory effect of short course administration of somatostatin (stilamin) continuously at early stage in patients with severe acute pancreatitis (SAP). METHODS: Thirty-nine patients with SAP (22 men, 17 women; the middle age was 49 years)were randomly allocated into control group (20 patients treated with non-surgical integrated traditional Chinese and Western medicine) and treatment group (19 patients treated with somatostatin administered intravenously at a dosage of 250 mug/h for consecutive 72 hours as well as the treatment for the control group). Laboratory parameters, including the expressions of CD(3), CD(4) and CD(8) in lymphocytes (tested by flow cytometry) and C reactive protein (CRP), and indexes of therapeutic effect, including the occurrence of organic dysfunction, local complication and mortality between the two groups were compared. Another group of 30 healthy volunteers (19 men, 11 women; the middle age was 47 years) were recruited for testing the normal levels of CD(3), CD(4) and CD(8). RESULTS: (1) The levels of CD(3), CD(4) and CD(4)/CD(8) in lymphocytes before treatment in both groups were significantly lower than those in the healthy subjects (P<0.05), but there were no statistical differences between the two groups. At the 4th day, CD(3), CD(4) and CD(4)/CD(8) increased significantly in the treatment group (P<0.05) while no changes in the control group; the levels of CD(4) and CD(4)/CD(8) in the treatment group were also higher than those in the control group (P<0.05). (2) The CRP levels of the 2 groups showed no statistical difference before and 4 days after the treatment, but the CRP level in the treatment group was significantly lower than that in the control group at the 7th day (P<0.05). WBC count, serum levels of amylase, lipase, lactate dehydrogenase, aspartate aminotransferase, as well as the score of APACHE II in the treatment group recovered more quickly than those in the control group (P<0.05). (3) The occurrences of organic dysfunctions, local complications and mortality in both groups were not statistically different. CONCLUSION: The short course administration of somatostatin continuously at early stage can reduce the inflammatory response, up-regulate the cell immune function and improve the conditions of the patients with SAP, but its effect on mortality and morbidity needs further study.

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.