Objective To observe the effect of T-2 toxin on growth and development of rat epiphyseal plate of left knee and metaphyseal bone of femur and tibia in normal and low nutritional status, to find out possible pathogenic factors of Kashin-Beck disease and provide experimental basis for early intervention. Methods Ninety 3-week-old Wistar rats, weighing 60 - 70 g, were randomly divided into three groups: control group(general feed), T-2 toxin + general feed group, T-2 toxin + low nutrition feed group, thirty rats in each group with equally sex ratio. T-2 toxin (1.0 mg/kg) was administered orally 5 times a week via a gavage needle for 4 weeks. The change of hair, activity and body weight was observed. After 1, 2, 4 weeks, the epiphyseal plate of left knee and metaphyseal bone of femur and tibia (including distal femur and proximal tibia) were collected. Specimens were processed with HE and Masson staining. The morphology of chondrocytes and matrix collagen content in epiphyseal plate was observed. Trabecular bone volume fraction in tibial metaphyseal bone was analyzed by Image-Pro Plus 6.0 software. Results In the control group, rats were in good movement and hair with light, but in T-2 toxin + general feed group and T-2 toxin + low nutrition feed group, rats were found with reduced activities and hair with dark color. Body weights(g) of the control group, the T-2 toxin + general feed group and the T-2 toxin + low nutrition feed group were 81.0 ± 6.2, 79.0 ±5.1, 77.0 ± 7.5, respectively, by the end of first week; 101.8 ± 6.7, 97.0 ± 6.8, 93.0 ± 5.3, respectively, by the end of second week; 151.1 ± 15.7, 126.5 ± 11.9, 106.5 ± 11.5, respectively, by the end of fourth week. There was significant difference in groups by second week and the fourth week (F = 9.72, 41.65, all P ＜ 0.05 ). There was significant difference among multi-groups by the fourth week(all P ＜ 0.01 ). Under light microscope, at the second weeks, coagulative necrosis of chondrocytes was found in hypertrophic zone in the two groups with T-2 toxin; at the fourth weeks, cell necrosis increased. Masson staining showed collagen staining in the two groups with T-2 toxin significantly turned to clear pale coloration, indicating that the collagen matrix was significantly reduced. Image analysis showed there was significant difference in groups at the second and fourth week(F= 9.72, 41.65, all P＜ 0.05)in tibial metaphyseal trabecular bone volume fraction. There was significant difference between T-2 toxin + low nutrition feed group[(0.55 ± 0.12)％, (0.21 ± 0.0)％] and control group[(0.67 ± 0.09)％, (0.51 ± 0.14)％] by the second and fourth week(all P ＜ 0.01 ). Conclusions Under normal nutritional status, T-2 toxin can induce hypertrophic epiphyseal cartilage necrosis, collagen content decreased in epiphyseal plate, metaphyseal trabecular bone formation disorders; in the low nutritional status, T-2 toxin can lead to rat epiphyseal necrosis and significant metaphyseal bone disorder, but whether the performance is related to Kaschin-Beck disease needs to be studied further.

Objective To investigate the effect of curcumin an extract of a Chinese medical herb on the sensitivity of CD133 + rectal cancer cells to radiotherapy.Methods In vitro experiments:CD133 +cells were purified with immunomagnetic beads from HRT-18 cell line and divided into curcumin group,radiotherapy group and curcumin plus radiotherapy group.MTT assay and Annexin V/PI staining were used to measure the proliferation and apoptosis of the cells.In vivo experiments:Transplanted rectal tumor was established in 46 nude mice and randomly divided into curcumin group,radiotherapy group and curcumin plus radiotherapy group.Tumor size and apoptosis were detected by daily observation and TUNEL staining respectively.Results Curcumin inhibited proliferation and apoptosis of CD133 + rectal cancer cells when combined with radiotherapy.It also significantly increased the growth inhibition of rectal tumor and promoted the apoptosis of rectal cancer in vivo.MTT assay showed that after 24 hours,compared with that of radiotherapy group(14.6％ ± 1.0％),curcumin plus radiotherapy group (18.7％ ± 1.7％) inhibited the growth of the tumor(P ＜ 0.01).Annexin V/PI showed that curcumin plus radiotherapy group (28.8％ ±3.7％) was significantly different from the radiotherapy group(13.1％ ± 1.4％) in cell apoptosis (P ＜0.01).In vivo,after 6 days,tumor volume (521 ± 79) mm3 in curcumin plus radiotherapy group was significantly lower than that of radiotherapy group(717 ± 134) mm3 (P ＜ 0.01) ; TUNEL staining results indicated that the RCST in curcumin plus radiotherapy group (26.1％ ± 3.3％) were higher than that in radiotherapy group (12.0％ ± 2.1％) (P ＜ 0.01).Conclusions Curcumin significantly enhances the radiosensitizing effect for CD133 + rectal cancer cells.

Objective To investigate the curative effect between liver resection combined with microwave ablation during operation and simple liver resection in the treatment of primary liver cancer.Methods From January 2005 to December 2013,a total of 84 patients diagnosed as primary liver cancer in our hospital were collected and divided into combination group(42 cases) and simple group(42 cases) according to the surgical method.Combination group were treated by combining liver resection with microwave ablation during operation,simple group by simple liver resection.Results The intraoperative blood loss for combination group was (323.9 ± 93.1) ml and simple group was (524.5 ± 119.2) ml,P ＜ 0.05.postoperative tumor recurrence rate for combination group was 14.2％ and simple group was 33.3％,P =0.040.1-,3-,and 5-year survival rate for combination group was 96.5％,67％ and 51％,and simple group was 84％,49.5％ and 36.5％,P =0.036.The differences of the above parameters between the two groups were statistically significant.The operation time for combination group was (177.7 ± 30.7) min and simple group was (165.1 ± 29.5) min,P =0.058.The postoperative hospital stay for combination group was (15.5 ± 3.7) d and simple group was (14.0 ± 4.0) d,P =0.068.The changes of ALT,AST,ALB,TBIL on the first postoperative day and the incidence of postoperative complications (including bile leakage,fever,pleural effusion,blooding,abdominal infection and some others) between the two groups had no statistical significances (P ＞ 0.05).Conclusion The curative effects of liver resection combined with microwave ablation during operation are superior to pure liver resection in the treatment of primary liver cancer.

The NSP2 gene of porcine reproductive and respiratory syndrome virus (PRRSV)S1 strain was partly amplified and cloned into a prokaryotic expression vector pGEX-6P-1 and a fusion protein GST-tNSP2 with molecular weight of 50 kDa was expressed in E.coli. The purified GST-tNSP2 protein showed a strong reaction with the PRRSV-positive sera in Western blot assay. Balb/c mice were immunized with the purified protein, and the splenocytes of the immunized mice were fused with murine myeloma cells SP2/0. After subcloning by 3 times, two hybridoma clones which produced McAbs steadily were screened by ELISA, named 3H3 and 2B5. They all reacted strongly with the PRRSV S1 infected Marc-145 cells in IFA, but not with the PRRSV SY0608 strain. Both of the McAbs belong to IgG1 isotype, and their light chains belong ? type. The expressed GST-tNSP2 protein and McAbs could be used for identification of PRRSV isolates and functional analysis of NSP2.

Objective:To express the EMCV 3AB gene by prokaryotic expression systerm,and prepare monoclonal antibodies against it. Method: NSP 3AB gene of Encephalomyocarditis virus (EMCV) was amplified and cloned into a prokaryotic expression vector pGEX-6P-1 and a recombinant protein 3AB with high antigenicity was expressed in E.coli. Balb / c mice were immunized by purified recombinant 3AB protein of inclusion-body, and the splenocytes of the immunized mice were fused with murine myeloma cells to produce hybridoma cell line. Results: After subcloning by 3 times, one strain of hybridoma cell line steadily secreting antibodies of 3AB protein was obtained, named 2D12. The McAb belongs to IgG1/?. The McAb and was confirmed by indirect immunofluorescent assay (IFA) and Western blot. Conclusion: These results can provide a potential value for structural and functional studies of EMCV-3AB and early diagnosis of Encephalomyocarditis virus infection.

Anemia ; etiology ; Erythema ; etiology ; Fatal Outcome ; Female ; Fever ; etiology ; Humans ; Hypereosinophilic Syndrome ; complications ; diagnosis ; Infant

OBJECTIVETo investigate the mechanism of hepatocytes transdifferentiation to bile duct epithelial cells (BECs) and intervention of Huangqi decoction (HQD) on hepatic fibrosis formation in rats with secondary cholestasis.

METHODSSeventy-five SD male rats were made into cholestatic hepatic fibrosis model animals by bile duct ligation, and randomized into the control group (n = 50) and the HQD group (n = 15). Starting from one week after modeling, they were administered orally with saline and HQD respectively for four weeks. Besides, a sham-operated group was set up with 10 rats operated by choledochus segregating only and administered after then with saline. Rats were killed in batches at different time points, i.e. each five from the control group and sham-operated group at the end of the 1st week, five from the control group for each time at the end of the 2nd, 3rd and 4th week, and all the remaining rats at the end of the 5th week. Their liver tissues were taken for histological change examination, content of hydroxyproline (Hyp) determination; protein expression of BECs marker cytokeratin 7 (CK7) and the hepatocyte specific antigen HepPar detection by Western blot, and CK7-Hep Par co-localization by laser confocal microscopy. Then IPP software was used to analyze Sirius red stained positive areas of CK7 and Hep Par, as well as the average IOD of CK7/Hep Par co-localization.

RESULTSHepatocytes in hepatic tissues (Hep Par positive cell) in the model rats decreased gradually along was time went by after modeling (Sham > M1w > M2w > M3w > M4w > M5w), which was in parallel with the increase of BECs (CK7 positive cells), degree of fibrosis, Hyp content and CK7 protein expression. Increasing of co-localized positive cells of CK7/Hep Par began at 1 week and reached the peak 3 weeks after modeling, then it decreased gradually. The Hep Par protein expression was negatively correlated with that of CK7; the Hep Par positive cell expression was negatively correlated with CK7 positive cell expression and collagen deposition; while the CK7 positive cell expression was positively correlated with the collagen deposition in the liver tissue. Compared with the model control group, the mortality, CK7/Hep Par co-localized positive cells, fibrosis degree, Hyp content and CK7 protein expression were lesser obviously (P < 0.01), while Hep Par positive cell and protein expressions were higher significantly in the HQD group.

CONCLUSIONSHepatocytes transdifferentiation to BECs might be a key pathological element for secondary cholestatic hepatic fibrosis formation; the restraining action of HQD is possibly a major action mechanism of HQD for effectively intervening and treating secondary cholestasis hepatic fibrosis.

Animals ; Astragalus Plant ; Bile Ducts ; cytology ; Cell Transdifferentiation ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Epithelial Cells ; cytology ; drug effects ; Hepatocytes ; cytology ; drug effects ; Liver ; Liver Cirrhosis, Biliary ; drug therapy ; pathology ; Male ; Phytotherapy ; Rats ; Rats, Sprague-Dawley

Adrenal Gland Neoplasms ; pathology ; surgery ; Adrenal Glands ; pathology ; surgery ; Child, Preschool ; Diagnosis, Differential ; Female ; Follow-Up Studies ; Granuloma, Plasma Cell ; pathology ; surgery ; Histiocytoma, Malignant Fibrous ; pathology ; Humans ; Neoplasms, Muscle Tissue ; pathology ; surgery

Objective To prepare the survival motor neuron(SMN)polyclonal antibody and explore the localization of SMN protein in transfected cells and its expression in skeletal muscles of patients with spinal muscular atrophy(SMA).Methods A prokaryotic expressional plasmid named pET-28? (+)/SMN was constructed and SMN-His fusion protein was induced.The fusion protein was used to immunize New Zealadd rabbits to prepare SMN polyclonal antibody.A eukaryotic expressional plasmid named pcDNA3.1/myc-HisB-SMN was constructed and used to transfect CHO cells.Skeletal muscles were collected from 3 patients with bone fracture who were regarded as normal controls, and 3 SMA patients of type Ⅰ, 3 of type Ⅱ and 3 of type Ⅲ who were ascertained by genetic analysis.Western-blotting and immunofluorescence stain were applied to study the expression of SMN in transfected CHO cells and skeletal muscles of normal individuals and SMA patients.Results Correct pET-28a(+)/SMN prokaryotic expressive plasmid was constructed and SMN-His fusion protein was obtained from E coli BL21 transformed with pET-28a(+)/SMN.Then, rabbit anti-human full-length SMN polyclonal antibody of high specificity and sensitivity was obtained from rabbits immunized by SMN-His fusion protein.SMN proteins were shown diffusedly locating in the cytoplasm and nucleus of CHO cells transfected with pcDNA3.1/myc-HisB-SMN plasmid and mainly accumulating around the nucleus.The results of Western-blotting were as follows:the average ratio of SMN band density to glyceraldehyde phosphate dehydrogenase(GAPDH)band density (SMN/GAPDH)is 0.619 in skeletal muscles from normal controls, the average values of SMN/GAPDH in skeletal muscle from SMA patients of type Ⅲ and Ⅱ were 0.347 and 0.340 respectively, which were lower than that of normal controls.However, the average values of SMN/GAPDH in skeletal muscle from SMA patients of type I was only 0.079, which was quite lower than that of normal controls.Conclusions The rabbit anti-human full-length SMN polyclonal antibody is of high specificity and sensitivity, which makes the basis for the research of SMN function and SMA pathogenesis.There may be a correlation between the SMN level in skeletal muscle and the severity of disease.

Objective To characterize the clinical course of biliary complications after right lobe living donor liver transplantation (RL-LDLT) and to identify the independent risk factors for biliary strictures.Methods 105 consecutive RL-LDLT recipients operated from April 2007 to April 2010 were followed up. The clinical and operative data were reviewed. The biliary complications and independent risk factors of biliary stricture were studied.Results The median follow-up duration was 49.5 months ranging from 562 to 1675 days.A total of 40 patients (38.1 ％) experienced 11 bile leak episodes (10.4％ ) and 37 (35.2％) biliary stricture episodes after transplantation.Bile leaks occurred at a median time of 9 days ranging from 4 to 54 days after transplantation.For biliary strictures,the occurring time was delayed and scattered wide with a median of 7.6 months ranging from 12 to 790 days after transplantation. Moreover, the biliary stricture incidence in the first year after transplantation was significantly higher than later.The independent risk factors for biliary strictures were CMV infection,bile leaks and bile duct size (≤3 mm).Conclusion The independent risk factors for biliary strictures after RL-LDLT were CMV infection,bile leaks and bile duct size (≤3mm).In order to avoid biliary complications,careful preoperative evaluations are necessary. The dissection of bile ducts should be meticulous to protect its blood supply.CMV infection should be prevented after transplantation.Close surveillance of biliary complications should be given to RL-LDLT recipients during the first year after transplantation.