BACKGROUND: Several studies have confirmed that activation of intervertebral disc enzymes is closely related to matrix degradation. Matrix metalloproteinase and tissue inhibitor of metalloproteinase have been shown to exert important roles in the process of extracellular matrix degeneration in intervertebrel disk. Besides these two enzyme systems, whether other proteeses that exhibit degrading effects on extracellular matrix are involved in the intervertebral disk degeneration remains poorly understood.OBJECTIVE: To detect the cathepsin K expression in normal and degenerated human intervertebrel disc cells and investigate the correlation between cathepsin K and intervertebral disc degeneration.METHODS: Cathepsin K expression was detected in intervertebral disc tissue from 30 patients with lumbar intervertebral disc protrusion using immunohistochemistry SP method and ELISA. At the same time, the intervertebral disc tissue from 15 healthy adult cadavers and/or spine fracture patients was taken as control, Cathepsin K protein expression in normal and degenerated human intervertebral disc tissues were compared.RESULTS AND CONCLUSION: Cathepsin K expression was observed in normal and degenerated intervertebral disc tissues.The expression level was significantly higher in degenerated tissue than normal tissue (P ＜ 0.05). These findings demonstrate that Cathepsin K possibly participates in the intervertebral disc degeneration.

Objective:To investigate memory disorder in hippocampal infarction patient. Methods: A hippocampal infarction patient was examined by the MMSE and the WMS-RC. Results: Patient's performance on MMSE was within the normal range, but the short-term memory was impaired. Conclusion: Hippocampal plays an important role in encoding and storing new information.

Objective To discuss causation and clinical treatment of complication in anterior decompression and fixation technique (ADFT) for thoracolumbar fractures with associated paraplegia. Methods A total of 204 cases that had thoracolumbar fractures with paraplegia were operated with ADFT from 1998 to 2004. Of all, 51 times (34 cases) of complication took place. Change of Cob angle was measured according to X-film of thoracolumbar spine before and after operation. Meanwhile, sensation and motion of patients was evaluated. Results There was incision infection leading to septic cerebo-meningtis in one case, CSF leakage in seven, pneumothorax in eight, rib nerve-root injure in five, lumbar nerve-root injure in two, genitofemoral nerve injure in three, lateral femoral cutaneous nerve injure in two, kyphosis in three, deep venous thrombosis in three and eight times of skin-temperature change. X-film that was taken after operation for all patients showed scolisis in five cases. A follow up for 3-6 years (average 2.5 years) was performed in 84 cases, of which seven cases had chronic low-back pain, six scolisis and five kyphosis. No patient showed complications such as screw breakage, plate breakage, fixation device loosening and pseudo-articulation in thoracolumbar spine. Conclusions Many complications can be avoided if we master well anterior decompression and fixation technique of thoracolumbar fracture, reduce surgical trauma and give appropriate and rational guide for rehabilitation.

Objective To analyze the nutrient constituents of the gonad from Hemicentrotus pulcherrimus.The results could provide a theoretical basis for the development of the H. pulcherrimus.Methods The contents of the water,ash and protein were determined by the national standard methods,total sugar by phenol-sulfuric acid method,rude fat by soxhlet's method,fatty acids by gas chromatography-mass chromatography,and inorganic elements by atomic absorption spectroscopy method.Results The contents of water,ash,rude fat, protein and total sugar were 64.20%,12.70%,2.34%,12.25%and 5.59%,respectively. The contents of arachidonic acid and EPA were higher in fatty acids.Inorganic elements, such as Ca,Mg,Fe were also in higher level.Conclusion The gonad of H.pulcherrimus was useful in the exploitation of nutrient food.

Objective To observe the protective effect of gradual ischemic postconditioning (IP) capable of improving reperfusion on reperfusion injury in patients with ST-segment elevation acute myocardial infarction (STEAMl) undergoing primary percutaneous coronary intervention (PPCI).Methods 102 in-patients with STEAMI undergoing PPCI in the Department of Cardiology in the First People's Hospital of Kunshan City Affiliated to Jiangsu University from February 2011 to August 2014 were enrolled in this study. They were divided into three groups by a random number table: IP group (32 cases), gradual IP group (30 cases) and routine reperfusion group (40 cases). In IP group, after the opening of the infarction related blood vessel, ischemic postconditioning within the first minute of arterial reperfusion was made through three episodes of 1 minute inflation and 1 minute pressure withdrawn of an angioplasty balloon, and then persistent reperfusion was carried out. In the gradual IP group, the patients received three times of gradual angioplasty balloon inflation and denation, 1 minute/1minute, 30 seconds/30 seconds and 15 seconds/15 seconds respectively, presenting the gradual change of IP time. In the routine reperfusion group, after the opening of blocked blood vessel, the patients underwent routine PCI to persistently recover the coronary artery blood supply. The changes of related lead ST segment regression (Sum-STR), incidence of reperfusion arrhythmia, corrected thrombolysis in myocardial infarction (TIMI) franle count (CTFC), peaks of MB isoenzyme of creatine kinase (CK-MB), left ventricular ejection fraction (LVEF) and frequency of adverse events in follow-up period were compared among the three groups.Results The baseline characteristics were comparable in three groups. The incidence of ventricular premature beats was significantly lower in gradual IP group than that in routine reperfusion group [30.0% (9/30) vs. 55.0% (22/40),P < 0.05], and although theincidence of ventricular premature beats was lower in IP group than that in routine reperfusion group [34.4% (11/32) vs. 55.0% (22/40)], no statistically significant difference was found (P > 0.05). The incidence of ventricular tachycardia was significantly lower in IP and gradual IP groups than that in routine reperfusion group [15.6% (5/32), 13.3% (4/30) vs. 40.0% (16/40), bothP < 0.05]. The incidences of ventricular fibrillation, bradyarrhythmia and sinus arrest were lower in IP group and gradual IP group than those in routine reperfusion group, but no statistically significant differences were found (allP > 0.05). In IP group and gradual IP group, the Sum-STR incidence, CTFC, CK-MB peaks were lower than those of routine reperfusion group [Sum-STR: (56.7±18.3)%, (57.3±21.5)% vs. (44.6±21.6)%; CTFC: 25.47±5.37, 24.46±6.41 vs. 31.62±7.56; CK-MB peaks (U/L): 126.3±78.5, 121.6±82.5 vs. 147.4±72.5; allP < 0.05], the effect of gradual IP group being the best among the three groups. The levels of LVEF were slightly higher in IP and gradual IP groups than the level in routine reperfusion group (0.507±0.042, 0.511±0.062 vs. 0.497±0.062), but no statistically significant difference was found (bothP > 0.05). In routine reperfusion group, one patient died because the ventricular fibrillation could not be corrected and another one died of no-reflow during operation. Each group had 1 patient died during the 4 weeks of follow-up after operation, in the routine reperfusion group, one died of refractory heart failure, and the cause of death of other two patients, one in IP group and another in gradual IP group, was considered due to subacute thrombosis in stent. Major bleeding events were not found in each group.Conclusion Gradual IP can ameliorate myocardial reperfusion injury more significantly in patients with STEAMI undergoing PPCI.

Objective To evaluate the safety,feasibility and efficacy of emergency percutaneous coronary intervention(PCI)on the spot in the patients with acute myocardial infarction(AMI)in country hospitals by interventional cardiologists from higher-level hospitals(converse transport).Methods A total of 81 AMI patients received emergency PCI on the spot by interventional cardiologists from other higher-level hospitals (transported doctors)from Mar 2004 to Sep 2008 in our hospital.The mean age of patients was 68.6 ± 3.6 years (36.0-83.0 years).Forty-six patients were male and 35 were female.There were 56 cases with anterior myocardial infarction and 25 with inferior myocardial infarction(including 11 cases combined with right ventricular infarction).The average time from symptom onset to admission was 6.2 ± 1.8 hours(2.0-12.0hours).Results Three cases were transported to higher-level hospitals for CABG because of severe conditions.The other 78 cases received emergency PCI on the spot,among whom 66 cases received primary PCI.Another 12 cases received rescue PCI.Eight one stents were implanted in total into the infarcted arteries.One operation was failed because the balloon could not go through the lesion.The success.rate was 98.7％.Four patients occurred peri-operative cardiac adverse events and 2 cases died.Four cases died during the 32-86 months follow-up,of whom I was cardiac death and 3 was non-cardiac deaths.No fatal cardiovascular events occurred in the remained cases.Conclusion Emergency PCI on the spot by interventional cardiologists from other cities(converse transport PCI)in AMI is safe,feasible and effective.But it needs to be confirmed in a large-scale study in the future.

Objective:To construct the Escherichia coli (E. coli)expression system for preparation of the bone morphogenetic protein-2 (BMP2)with collagen-binding domain (CBD),and to study the methods and conditions for expression, purification and renaturation of CBD-BMP2.Methods:CBD sequence was cloned into the N-terminal of BMP2 sequence, the recombinant vector pet21b/CBD-BMP2 was constructed and transformed into E.coli BL21.The expression of recombinant protein was induced using isopropylβ-D-thiogalactopyranoside (IPTG) at 37 ℃.Ni-NTA chelate chromatography was used to purify CBD-BMP-2.Denaturing CBD-BMP2 was refolded by dilution method using ultrapure water.The refolding CBD-BMP2 was filtered through a 0.22μm microfiltration membrane for degermation.The recovery rate was calculated by the ratio of the protein concentration before and after degermation. The expression, purification, and renaturation of recombinant protein were detected by SDS-PAGE method.The concentration of CBD-BMP2 was detected by BCA assay.Results:The recombinant vector pet21b/CBD-BMP2 was successfully transformed into E.coli BL21,and the recombinant protein was expressed as inclusion bodies in E.coli.The SDS-PAGE results showed denaturing protein was dissolved in supernatant of lysis buffer with 8 mol·L-1 urea and the purified recombinant protein existed in elution buffer B with relative molecular mass about 14 000.Two bands (14 000 and 28 000)were seen in the SDS-PAGE picture,which indicated that the monomer was successfully refolded into dimer by dilution method.The concentrations of recombinant protein before and after degermation were 110 and 80 mg · L-1 , respectively, and the recovery rate was about 73%. Conclusion:The recombinant vector pet21b/CBD-BMP2 is transformed into E.coli BL21 successfully,and the recombinant CBD-BMP2 is expressed and refolded efficiently. The methods of prokaryotic expression system for preparing recombinant CBD-BMP2 protein are established.

Objective To study the effect of positive lymph node number on the survival of patients with esophageal carcinoma. Methods From July 1995 to July 2005, a total of 11,447 resected lymph nodes were obtained from 1140 patients who underwent curative resection of the primary tumor with systematic lymphadenectomy at Shanxi cancer hospital. The survivals were analysed by life tables and Kaplan-Meier methods, the related factors of lymph node metastasis were assessed by Chi-square test. Results The number of positive lymph nodes was negatively related to survival rates of esophageal carcinoma. According to the number of lymph nodes resected (≥8 nodes versus <8 nodes), there was significant difference in metastatic lymph node ratio. Conclusion The number of positive lymph node can reflect the prognosis of patients better. The authors suggest that the modification of the tumor-lymphnode-metastasis (TNM) staging classification (TNM) to include the number of positive lymph nodes in the N1 category.

Objective To explore the expression, correlation with clinicopathologic parameters, and clinical significance of MIS18 binding protein 1 (MIS18BP1) in bladder cancer. Methods TCGA and GEO databases were used to analyze the mRNA expression of MIS18BP1 in tumors and controls, and the results were verified via qRT-PCR. UALCAN online database was utilized in the analysis of the expression of MIS18BP1 and its correlation with clinicopathological parameters and the degree of immune cell infiltration. Immunohistochemistry was employed to analyze the expression of MIS18BP1 in bladder cancer and its relationship with clinicopathological features. The ROC curve was applied to evaluate the diagnostic value of MIS18BP1 mRNA in bladder cancer. Results Bioinformatics analysis and qRT-PCR results revealed the increased expression of MIS18BP1 mRNA in bladder cancer compared with that in the control group (P<0.05). Immunohistochemistry unveiled the significantly high positive rate of MIS18BP1 protein in bladder cancer (P<0.05) and its correlation with the clinical stage of tumors, depth of invasion, and lymph node metastasis (P<0.05). The immune infiltration analysis showed the association of MIS18BP1 with immune cell infiltration in bladder cancer. Conclusion The increased expression level of MIS18BP1 gene and protein in bladder cancer may regulate the development of bladder cancer by influencing immune cell infiltration.

cry1Ah1, one of holo-type cry genes, cloned in this laboratory from Bacillus thuringiensis strain has been patented in China, and it encoded a protein with strong insecticidal activity against certain lepidopteran insect pests, such as Chilo suppressalis. cry1Ah1 gene is exhibiting good application prospects. In order to improve the expression level of cry1Ah1 gene in rice, and investigate the effect of codon usage preference of gene expression, we designed five different optimized schemes for cry1Ah1 insecticidal critical fragment in accordance with bias of rice codon, to improve G+C content, removed the shear signal and unstable factors. Optimized cry1Ah1 genes were transformed into Escherichia coli Rosetta (DE3) respectively, and 65 kDa polypeptides was expressed normally in inclusion body separately. All of these expressed polypeptides showed insecticidal activity against 2nd-instar larvae of Plutella xylostella and neonate of Chilo suppressalis. After transformation with modified cry1Ah1 genes into Var nippobare, the transgenic rice seedlings were detected by PCR, the positive rate containing target gene was more than 87%. Afterwards, the results of real-time RT-PCR and ELISA assay indicated that the highest expression level of five modified cry1Ah1 genes was that using the highest frequent codons. Average expression amount of Cry1Ah1 polypeptides was 0.104% of total soluble proteins from the positive transgenic rice.
Animals ; Bacillus thuringiensis ; genetics ; metabolism ; Bacterial Proteins ; biosynthesis ; genetics ; Cloning, Molecular ; Codon ; genetics ; Endotoxins ; biosynthesis ; genetics ; Hemolysin Proteins ; biosynthesis ; genetics ; Insecticides ; Lepidoptera ; Oryza ; genetics ; Pest Control, Biological ; methods ; Plants, Genetically Modified ; genetics ; Recombinant Proteins ; biosynthesis ; genetics