AIM:To observe the expressions of cathepsin B (CB) and cystatin C (CC) in different stage of diabetic rats and to investigates their potential roles.METHODS:Sixty rats were divided into diabetes mellitus group induced by intravenous injection of streptozotocin (55 mg/kg) and normal group injected with citrate buffer. Ten rats were sacrificed respectively at the end of fourth week,eighth week and sixteenth week in both groups. 24 h urine excretion was collected in rats before sacrifice. The blood and the kidney were also collected. The mRNA and protein expressions of CB and CC in kidney were detected by real time PCR and immunohistochemical staining,respectively.RESULTS:At the end of eighth week,the expression of Ccr,24 h urinary protein excretion,CB,CC in diabetic rats increased significantly,compared to the results at the fourth week (P

OBJECTIVE To analyze the profile of the pathogens and their drug resistance isolated from children with lower respiratory tract infection in Wenzhou area from Jan 2004 to Dec 2006.METHODS Lower respiratory tract secretions were obtained from children with lower respiratory tract infection for bacterial culture.The K-B method was applied for the antibiotic susceptibility test.RESULTS Total 1605 strains were isolated.The isolating rates of Gram-positive cocci,Gram-negative bacilli and fungi were 24.9%,61.2% and 14.0%,respectively.60.5%,and 54.6% of the isolates of Escherichia coli and Klebsiella pneumoniae produced extended-spectrum beta-lactamases(ESBLs).The rate of Penicillin-resistant Streptococcus pnenmoniae(PRSPN)was 30.6%.20.5% isolates of Staphylococcus aureus were MRSA.All isolates of Enterobacteriaceae and Gram-positive cocci were susceptible to imipenem and vancomycin in vitro.CONCLUSIONS Gram-negative bacilli are still the primary pathogens resulting in lower respiratory tract infection in children.Fungi and muti-drug-resistant bacteria are on the rise trend.

To study the effect of cholesterol and 25-OH-cholesterol on cholesterol metabolism in HepG2 cells and the effect of coptisine (Cop) extracted from Coptidis Rhizoma (CR) in reducing and regulating cholesterol. In this study, TC, TG, LDL-c and HDL-c were measured by biochemical analysis; mRNA and protein expressions of LDLR, HMGCR and CYP7A1 were detected by qRT-PCR and Western blot. According to the results, cholesterol and 25-OH-cholesterol inducing could decrease in mRNA and protein expressions of LDLR and CYP7A1, so as to increase TC and LDL-c contents. However, Cop could up-regulate mRNA and protein expressions of LDLR and CYP7A1 and down-regulate that of HMGCR, so as to reduce TC and LDL-c levels. These findings suggested that Cop has potential pharmacological activity for reducing cholesterol, and may reduce cholesterol by regulating mRNA and protein expressions of key genes involved in cholesterol metabolism, such as LDLR, CYP7A1 and HMGCR. This study laid a firm theoretical foundation for developing new natural drugs with the cholesterol-lowering activity.
Berberine ; analogs & derivatives ; pharmacology ; Cholesterol ; metabolism ; Cholesterol 7-alpha-Hydroxylase ; genetics ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Gene Expression Regulation, Enzymologic ; drug effects ; Hep G2 Cells ; Humans ; Hydroxymethylglutaryl CoA Reductases ; genetics ; metabolism ; Receptors, LDL ; genetics ; metabolism ; Triglycerides ; metabolism

ObjectiveTo explore the effect of different nutritional support mdoes on humoral immunity and outcomes after esophagectomy in patients with esophageal carcinoma.MethodsForty-six patients with middle or low thoracic esophageal carcinoma underwent Ivor Lewis esophagectomy.The patients were randomized into enteral nutrition group ( EN,n =23 ) and enteral combined parenteral nutrition group ( EN + PN,n =23 ) based on the nutrition support modes.Serum levels of immunoglobulin (IgG,IgA,IgM,IgE,κ/λ light chain) and comphments (C3/C4) were assayed and compared on the 1st pre-operative day and at 18 hours as well as 3rd and 7th day after operation.The clinical outcomes including infection-related complications and hospital stay were compared between two group s.ResultsThere was no significant difference in all humoral immunity indicators between two groups at the eachpost-operative time point.In both two groups,the levels ofIgG [ (8.90 ± 1.75),(7.53 ±1.41) g/Land (8.64±2.44),(7.48±2.16) g/L],κ [ (2.14±0.46),(1.78±0.41) g/L,and (2.15 ±0.63),( 1.86 ± 0.62) g/L] and λ light chain [ ( 1.34 ± 0.45 ),( 1.11 ± 0.31 ) g/L and ( 1.20 ± 0.32),( 1.08 ± 0.35 ) g/L] were significantly lower 18 hours and 3rd day after operation than the pre-operative levels [ (12.15±2.86)and (11.11±2.96) g/L,(2.90±0.77) and (2.77±0.79) g/L,(1.79±0.57) and (1.56±0.41) g/L] (P=0.000,P=0.000,and P=0.004,P=0.000,and P=0.000,P=0.000,and P=0.011,P=0.000,and P=0.004,P=0.000,and P =0.008,P =0.000),and returned to the preoperative levels by the postoperative 7th day (P＞0.05),except for the level of κ light chain 7th day after operation in EN group [ ( 2.42 ± 0.69) g/L] ( P =0.027 ).The levels of IgA,IgE,and C3 were not significantly different during the perioperative period ( P ＞ 0.05 ).The level of IgM was not significantly different during the perioperative period in EN group (P ＞0.05),and was significantly lower on the 3rd post-operative day [ ( 1.00 ±0.53) g/L] than the pre-operative level [ ( 1.47 ±0.76) g/L] in the EN + PN group (P =0.031 ),and were not significantly different on the other time points (P ＞ 0.05 ).In the EN group,the C4 level was significantly lower at the postoperative 18 hours [ (0.24 ±0.08) g/L] than the pre-operative level [ (0.37 ±0.36) g/L] (P =0.030),and were not significantly different at the other time points ( P ＞ 0.05 ).In the EN + PN group,the C4 level was not significantly different during the perioperative period ( P ＞ 0.05 ).There was no significant difference in the infection-related complications and hospital stay between these two groups ( P =0.300,P =0.371 ).ConclusionsThe effects of EN or EN + PN on humoral immunity and outcomes after esophagectomy in patients with esophageal carcinoma are not different.Both these two nutritional support modes can not completely alleviate the harm to the humoral immunity.The EN is more cost-effective.

Objective To observe the effects of transplantation of autologous adipose-derived stem cells (ASCs) on osteoporosis (OP) in a rabbit ovariectomy (OVX) model.Methods A total of fifteen 6-month-old female New Zealand white rabbits were randomly divided into two groups:ovariectomy group (group A,n =12) and sham operation group (group B,n =3).All rabbits were subjected to bilateral ovariectomy in the group A.Six months later,bone mineral density (BMD) of group A and group B were measured by dual energy X-ray absorptiometry (DXA) to check the result of OVX-OP.ASCs harvested from adipose of OP rabbits were cultured to be expanded and differentiated in osteogenic medium in vitro.Osteogenesis was evaluated by alizarin red staining,alkaline phosphatase (ALP) staining and quantitative assays of osteocalcin (OCN).Autologous osteo-induced ASCs were mixed in calcium alginate hydrogel (CAH) and then transplanted in the left distal femurs,while CAH was transplanted in the right distal femurs of OP rabbits.At 12 weeks after implantation,BMD,micro-CT and histomorphological analyses were performed on these rabbits.Results The BMD of femurs in group A rabbits were obviously lower than that of group B rabbits (P ＜ 0.05) at 6 months after OVX.Compared with control group,ASCs cultured in osteo-induction medium had similar proliferation rate as the non-induced cells,but displayed positive ALP and alizarin red staining and OCN contents.At 12 weeks after implantation,the cell-treated femurs displayed higher BMD,bone trabecula number,trabecula thickness and separation than those of control group,while the structure model index and porosity were lower (P ＜ 0.05).Histological examination indicated that the trabecular thickness increased with complete CAH resorption in cell-treated group,while CAH remained in control group.Conclusions Transplantation of autologous ASCs can help strengthen osteoporotic bone in OVX-OP rabbits,providing a novel approach to OP treatment.

Objective To investigate mutation and deletion of genes mecR1 and mecI in clinical methicillin-resistant staphylococci isolates and study the mutation and deletion have effect on gene mecA expression and drug resistance phenotype.Metheods PCR was used to detecte gene mecA and the regulatory genes mecR1 and mecI in staphylococci which were separated from clinical specimen in 2006,then the sequence of gene mecI was determined and compared with the sequence obtains from pre-MRSA strain N315(GI:BA000018).Results Gene mecA was detected in 60 strains of Staphylococcus aureus,58 strains of Staphylococcus epidermidis and 37 strains of Staphylococcus heamolyticus,but gene mecA in 6 strains of Staphylococcus epidermidis and 4 strains of Staphylococcus heamolyticus were only amplified by primer mecA2-F/R and not by primer mecA1-F/R.The percentage of gene mecR1 exist in Staphylococcus aureus was higher than Staphylococcus epidermidis and Staphylococcus heamolyticus,but the percentage of gene mecR1 exist in Staphylococcus epidermidis was not higher than Staphylococcus heamolyticus.The mutation and deletion of gene mecI were often seen,the wild type mecI was only detected in 14 strains,the point mutation of nucleotice 202 was detected in 36 strains.Conclusions Gene mecA expression in Staphylococcus aureus could be chiefly induced by mecR1,but which in coagulase-ngeative staphylococci could be other factors.The mutation and deletion of mecI were universal phenomenon in clinical strains,there could be a mechanism for overcoming the repressing of resistance caused by mecI in staphylococci.

Objective To predict the secondary structure and the B cell epitopes of human heparanase protein, and to identify its immunogenicity. Methods The flexible regions of secondary structure and the B cell epitopes of human heparanase amino acid sequence were predicted by DNAStar and Bcepred software. The multiple antigenic peptides (MAP) of the epitopes were synthesized in 8-branch form. Rabbits were immunized with the 8-branch MAPs mixed with a universal T-helper epitope human IL-1β peptide (VQGEESNDK, amino acid 163-171 ). The immunogenicity of the synthesized peptides was evaluated by ELISA, Western blot and immunohistochemistry. Results Amino acid 1 -15 ( MAP1), 279-293 (MAP2) and 175-189(MAP3) of large-subunit of human heparanase protein was predicted as the most potential epitopes of human heparanase protein. All the three synthesized MAPs induced high titer of antibodies. ELISA, Western blot and immunohistochemistry analysis showed all the three MAPs could produce high titer serum antibodies, antibodies induced by MAP1 and MAP2 had high specific binding activity , and MAP2 antibody showed the strongest binding activity with liver cancer tissues. Conclusion The large-subunit No. 1-15, 279-293 amino acid of human heparanase protein may be the B cell preponderant epitopes and the strongest immunogenicity may be No. 279-293 peptide, which provided a theoretic basis for the antibody and vaccine development of heparanase subunit peptide.

Objective The aim of this study was to investigate the differences on origin distribution and radiofrequency ablation characteristics between premature ventricular contractions (PVC) from left and right ventricular.Methods A total of 354 frequent PVC patients were enrolled.Two hundred and eighty patients were PVCs with right ventricular origin (group RV) and 74 patients with left ventricular origin (group LV).The age,gender,ablation power,temperature and duration,time of target potential before QRS onset,fluoroscopic time,procedure time and success rate were compared between the two groups.Results Majority PVCs (79.10％) were of right ventricular origin,in which the most common site was at right ventricular outflow tract (RVOT) (55.93％);whereas,only 20.90％ were of left ventricular.There were significantly differences between group RV and group LV in age [(44.7 ± 13.6) yrs vs (49.6 ±15.9) yrs,P =0.017],gender [male:35.4％ (99) vs 55.4％ (41),P =0.002],time of target potential before QRS onset [(37.9 ± 16.2) msvs (31.3 ± 11.7) ms,P=0.008],procedure time [(85.1 ± 36.9) minvs (100.8 ± 45.5) min,P=0.017],fluoroscopic time [(12.3 ± 9.2) min vs (15.9± 10.8) min,P =0.028] and success rate (96.4％ vs 89.2％,P =0.012).Conclusions Radiofrequency ablation of PVCs was a safe and effective method.Subjects with RV PVCs are younger and more females than subjects with LV PVCs.The radiofrequency ablation for RV PVCs was easier,and the success rate was higher than that for LV PVCs.

Adult ; Humans ; Male ; Obesity Hypoventilation Syndrome ; diagnosis ; therapy ; Polycythemia ; diagnosis ; therapy

Objective To investigate the clinical, laboratory and genetic features of glycogen storage disease(GSD)IXc. Methods Five patients suspected as liver GSD were included in our study. DNA was extracted from peripheral blood of all the patients and diagnoses were made after target sequencing to nearly 2700 disease causing genes. All detected mutations were confirmed in the probands and their parents. Further analysis was based on clinical features, routine laboratory examinations and treatment. Results All the 5 patients manifested with severe hepatomegaly, hypoglycemia, moderately to severely elevated liver enzyme levels, hypertriglyceridemia and growth retardation. Four cases showed poor exercise tolerance but with normal creatine kinase (CK) levels. None of the patients showed liver cirrhosis. Growth velocity and hepatomegaly was improved after the uncooked corn starch treatment was initiated. In the 5 patients, 6 different pathogenic or likely pathogenic mutations in the PHKG2 gene were identified, including one reported mutation (p.E157K) and five novel mutations (p.E56X, p.R185X, c.79_88delinsTCTGGTCG, c.761delC,p.R279C). The p.E157K was the most frequently mutation identified (6/12, 50%). Conclusions The p.E157K mutation is the hot mutation in our small cohort. Main clinical features of our patients include fasting hypoglycemia, impaired liver function,short statures and poor exercise tolerance, without developing liver cirrhosis.