Objective To analyze the drug therapeutic regimen and duration of the chemotherapy in the spinal tuberculosis in order to determine the best therapeutic regimen and duration of the spinal anti-tuberculo-sis. Methods The medication plan and duration of 890 hospitalized patients with spinal tuberculosis in our hospital from January 2001 to December 2008 were retrospectively investigated. We collected the plan of the initial treatment,retreatment,recrudescence and drug resistance. We also studied the individuation therapeutic regimen of the patients with tuberculosis at other parts or complications. On the basis of these data,we analyzed the interferential appearance that the antituberculosis drug acted on different onsets,manifestations and operations. Results In 890 patients of spinal tuberculosis,596 cases ( 67% ) were initial treatment,294 ( 33% ) were retreatment,recur cases were 110 ( 12. 3% ) ,drug resistance cases were 74( 8. 3% ) ,and those compli-cated with tuberculosis in other parts were 273 ( 30. 7% ) . The main chemotherapeutic regimen was the usual tetragenous protocol ( H12 /R12 /E9 /Z5 /S3) . The retreatment cases were given second-line drugs such as levo-floxacin. The drug resistance cases were also given second-line drugs and intravenous infusion. According to the anti-tubercle bacillus spectrum,individuation treatments were adopted,and the plan was changed in time. The medication duration continued for 9-36 months. All the 890 patients were cured. Conclusion For the treatment of spinal tuberculosis,it is significance to make a efficient and standardized anti-tuberculosis chemotherapy.

Objective To observe the mitochondrial damage associated with protein-energy wasting of skeletal muscle in diabetic kidney disease (DKD) model of Goto-Kakizaki(GK) rats and evaluate the effects of low-protein diet supplemented with α-keto acids on muscle wasting.Methods Forty-five male 24-week-age GK rats were randomly divided into three groups,normal protein diet group (NPD),low-protein diet group (LPD) and LPD +or-keto group (Keto).Fifteen gender and age matched Wistar rats were served as control group (CTL).The living condition of GK rats was observed and the weight was measured once a week.Urine albumin,serum glucose,creatinine and urea nitrogen were measured at 24,32,40,48 week age.Soleus muscle was observed to calculate the muscle size and the percentage of Ⅰ and Ⅱ type muscle fiber with software after SDH and NADH staining at 48-week-age.Tissue ultrastructure was observed under the transmission electron microscopy.The activity of citrate synthase was detected by spectrophotometer.Expression of mitochondrial DNA was examined by Q-PCR.Results Compared with the CTL group,NPD,LPD and Keto groups had lower body weight,higher urine albumin,higher serum creatinine and urea nitrogen (P ＜ 0.05).The crosssectional area of muscle fibers was larger in CTL group.Compared with CTL group,the muscle fiber was partly broken,the mitochondrial morphology was obviously changed,the percentage of type Ⅱmuscle fiber was increased significantly (P ＜ 0.05),and the activity of citrate synthase and the number of mitochondrial DNA were decreased significantly in NPD,LPD and Keto groups (P ＜ 0.05).In Keto group,muscle wasting was improved compared with NPD and LPD group (P ＜ 0.05),the crosssectional area of soleus muscle increased and the percentage of type Ⅱ muscle fiber decreased,levels of urine albumin,semm creatinine and urea nitrogen decreased (P ＜ 0.05).Under transmission electron microscopy,the muscle fiber of keto group was intact and mitochondiral morphology was close to that of CTL group.The activity of citrate synthase and number of mitochondiral DNA were higher as compared to CTL group (P ＜ 0.05).There were no significant differences between NPD and LPD group.Conclusions In DKD condition,protein degradation in the skeletal muscle is accelerated,mitochondrion is swelling,the number of mitochondrial DNA is decreased and mitochondrial function is impaired.Low-protein diet supplemented with α-keto acids can improve mitochondrial damage and muscle wasting induced by DKD.

Objective To investigate the causes and treatment strategies of spinal multidrug-resistance tuberculosis.Methods Data of 16 patients with spinal multidrug-resistance tuberculosis from Jane 2007 to September 2012 were retrospectively analyzed.There were 12 males and 4 females,with an average age of 26.6 years (range,10-49 years).The 16 patients involved 44 vertebrae,with an average of 2.75 vertebrae.The involved segments included:9 thoracic segments,1 thoracic-lumbar segment,2 lumbar segments and 3 lumbar-sacral segments.1 patient involved jumping segments including T8.9,T12L1.Among them,5 suffered from pulmonary tuberculosis,4 tuberculous pleurisy,3 tuberculous empyema,1 tuberculosis of cervical lymph nodes,1 tuberculosis of sternum,1 tuberculosis of chest wall and 1 nephrotic syndrome.We analyzed the reasons of multidrug-resistance.All patients received individualized chemotherapy based on drug sensitivity test.The operation process and time were also collected.The treatment effects were determined by long-term follow-up.Results Among all the 16 patients,6 received 1 operation; 7 received 2 operations; 2 received 3 operations and the last operation was one-stage posterior instrumentation and anterior debridement,bone grafting which conducted in our hospital; 1 received 4 operations and the last of which was excision of sinus in our hospital.All patients were followed up for 10 to 60 months (average,28.4 months).The time of chemotherapy which accorded to the drug sensitivity test was 24 months.2 cases recurred after 22 months and 46 months of the 1st surgery and received operation again.At the last follow-up,all patients were in a stable state of tuberculosis.In 16 patients,2 were initial drug resistance and 14 were acquired drug resistance.The causes of acquired drug resistance were multiple organs tuberculosis caused by failure chemotherapy,times of failed surgeries without adjusted schedules,suspension of the anti-tuberculosis chemotherapy due to serious adverse drug reactions and so on.Conclusion It is very important to carry through the culture of tubercle bacillus and acquire the results of drug sensitive test earlier.The key to prevent and cure multidrug-resistant tuberculosis of spine are formulating individualized anti-tuberculosis chemotherapy program,monitoring closely the adverse drug reactions and selecting the appropriate time for surgery.

Objective To investigate the incidence and associated factors of pre-eclampsia in women with abnormal glucose metabolism in pregnancy. Methods A retrospective study was conducted on 1202 pregnant women with abnormal glucose metabolism who delivered their babies in our hospital between 1981-2003. All women were divided into 2 groups: group Ⅰ included 151 women with pre-eclampsia; group Ⅱ consisted of 1050 women without pre-eclampsia. The risk factors of pre-eclampsia were analyzed. Results (1)The incidence of pre-eclampsia was 12.6% as a whole and was 34.8%(39/112), 11.8%(89/753) and 6.8% (23/337) in diabetes mellitus(DM), gestational diabetes mellitus(GDM) and gestational impaired glucose tolerance(GIGT) groups, respectively ( P

Objective To observe the muscle wasting in diabetic kidney disease (DKD) model of type 2 and non-obese diabetes mellitus in Goto-Kakizaki (GK) rats,and to evaluate the effect of lowprotein diet supplemented with α-keto acids on muscle wasting.Methods Forty-five male 24-weekage GK rats were randomly divided into three groups:normal protein diet group (22％ casein diet,NPD),low protein diet group (6％ casein diet,LPD) and LPD + α-keto group (5％ casein + 1％ α-keto,Keto).Fifteen gender-and age-matched Wistar rats were served as the control group (CTL).The living condition of GK rats was observed and body weight was measured once a week.Urine albumin,serum glucose,lipids,albumin,creatinine and urea nitrogen were measured at the age of 24,32,40,48 weeks.Soleus muscle at the age of 48-week was observed to calculate the muscle size with software.Expressions of atrogin-1,MuRF-1 and MyoD,myogenin were examined by Q-PCR and Western blotting.Results Compared with the CTL group,NPD,LPD,Keto groups had lower body weight [(317.90± 13.81),(330.38±11.96),(390.44±12.25) g vs (429.43± 16.85) g,all P ＜ 0.05],higher urine albumin [(14.36±5.52),(8.12±4.61),(5.58±3.50) mg/24 h vs (0.61±0.16) mg/24 h,all P ＜ 0.05],higher serum creatinine [(81.50±7.88),(66.32±8.36),(63.44±8.21) μmol/L vs (24.43±6.15) μmol/L,all P ＜0.05] and urea nitrogen [(7.53±1.05),(5.63±1.40),(5.54±0.97) mmol/L vs (2.98±0.62) mmol/L,all P ＜0.05].The cross-sectional area of soleus muscle fibers was larger in CTL group.Compared with CTL group,the expression levels of atrogin-1 and MuRF-1 increased significantly (all P ＜ 0.05),and of MyoD and myogenin decreased significantly in NPD,LPD,Keto groups (all P ＜ 0.05).In Keto group after 40 weeks,muscle wasting was improved compared with NPD and LPD group [body weight (381.62± 15.82) g vs (331.50±17.58),(326.60± 13.43) g,all P ＜ 0.05],cross-sectional area of soleus muscle increased,levels of urine albumin,serum creatinine and urea nitrogen decreased (all P ＜ 0.05),the protein expressions of atrogin-1 and MuRF-1 decreased,and myogenin and MyoD were higher as compared to CTL group (all P ＜ 0.05).There were no significant differences between NPD and LPD group.Conclusions In DKD condition,protein degradation in the skeletal muscle is accelerated,the genes which control muscle atrophy are activated,and proliferation and differentiation of the muscle satellite cells are impaired.Low-protein diet supplemented with α-keto acids can improve muscle wasting induced by DKD.

ObjectiveTo study the clinical features of the bilateral frontal brain contusion with cerebral hernia center and its treatment strategies. MethodsThe clinical data of 76 patients with cerebral central hernia were restropectively analyzed. ResultsIn 76 patients,there were 53 cases survive,23 cases died.The life and survival quality of these patients were evaluated according to the Karnofsky scale systerm :46 patients underwent surgery,including 35 cases with good recovery,8 cases with long-term coma or unable to look after themselves,3 cases with death;10 cases underwent a expectant treatment,of which,6 cases with good recovery,4 cases with long-term coma or unable to look after themselves.The other 20 cases died of central brain stem failure,with a central hernia when admissioned. ConclusionPatients with bilateral frontal brain contusion were extremely complicated with central hernia,and had a suddenly deteriored condition.Close observation of changes were critical importance.The surgery should be carried out before "diencephalon period",for most recovery well after surgery.Patients with diffuse brain swelling should go under the depressioning surgery as earlier as possible.

Objective To generate recombinant Pichia pastoris for high-copy expression of human tissue factor pathway inhibitor (TFPI). Methods The cDNA encoding human TFPI was inserted into the expression vector pPIC9K and the constructed expression vector rhTFPI-pPIC9K was confirmed by restriction endonuclease analysis and DNA sequencing. The recombinant plasmids were subsequently transformed into Pichia pastoris GS115 cells, and the transformants were confirmed by PCR amplification of the genomic DNA.The recombinant Pichia pastoris with high copies of TFPI cDNA was screened by G418 selection. Western blot and TFPI ELISA Kit were employed to analyzing. The temperature, time and concentration of methanol for the induction of recombinant protein were optimized. Results PCR analysis and DNA sequencing confirmed the successful construction of the expression vector rhTFPI-pPIC9K. Real time quantitative PCR and Western blot analysis demonstrated the positive correlation between TFPI expression level and the copy number of TFPI cDNA in Pichia pastoris cells. Optimization of the induction condition significantly elevated the expression level and activity of TFPI (9.95±0.78 mg/L and 3.91±1.37 U/mL). Conclusion The Pichia pastoris strain with high copy of TFPI expression was successfully constructed, which lays a solid foundation for the further investigation on the function of TFPI and its application in the prevention and therapy of diseases.

Objective To investigate the immunosuppressive regimen of cyclosporine A(CsA) reduced or withdrawn in the HBV-DNA positive kidney transplanted patients. Methods The program of 64 kidney transplanted patients with HBV-DNA positive from Jan,2004 to Dec,2007 were analyzed, the patients were divided into 3 groups ①CsA + MMF group(A group) ;②FK506 + MMF group(B group) ;③low dose of CsA + SRL group(C group). All the patients received entecavir to resist HBV replication and were followed up for acute rejection incidence,liverfunc- tion and HBV-DNA test for 6 months. Results There was no significant difference in 3 groups about acute rejection incidence rate. Liver dysfunction took place in 12 patients of A group (80%) ,8 patients(53%) in A group HBV-DNA became negative; 5 patients (20%) in B group appeared the liver dysfunction, HBV-DNA became negative in 18 patients(75%). 4 patients in C group(16%) appeared liver dysfunction sHBV-DNA was negative in 18 patients (72%) of C group. Conclusion It was safe and efficient for the immunosuppressive regimen of cyclosporin A re-duced or withdrawn in the HBV-DNA positive kidney transplanted patients,not increasing the incidence of acute re-jection and aggratating the liver injury.

Objective To investigate the effects of donor-specific regulatory T cells (Treg) transfusion on islet allograft survival. Methods Allogeneic fresh islets from Balb/c mice were transplanted to streptozotocin-induced diabetic C57 mice. The survival of islet allografts was observed. The experiment was divided into 3 groups: control group, nothing had been done to the recipients; simple islet transplantation group, the recipients received the islet transplantation only; experimental group, the recipients were given 1 ×106 Treg, then received islet transplantation. Results Blood glucose (BG) was above 16. 7 mmol/L after islet transplantation in control group; In simple islet transplantation group,BG level returned to normal level 1 to 2 days after transplantation, and hyperglycemia appeared 7 to 11 days after transplantation and maintained as the same as that before transplantation; In experimental group, BG level returned to normal level 2 days after transplantation and maintained at a low level,and at the 21st day after transplantation BG level was over 16. 7mmol/L in some recipients. Islet allograft survival in experimental group was significantly prolonged as compared with simple islet transplantation group. Conclusion Donor-specific Treg transfusion could prolong the islet allograft survival,and maybe have positive effect on tolerance induction of islet transplantation.

Objective:To investigate RNA interference and apoptosis induced by LPS in kidney epithelial cells through silencing C5aR gene with small hairpin RNA (shRNA).Methods:We construct the eukaryotic expression vector of small hairpin RNA targeting rat C5aR gene,and transfected RK3E cell by electroporation,after G418 selection,so we got the stable cell line expressing C5aR shRNA.The experiment was designed into 3 groups:①normal control group,RK3E cells without transfection;②negative control group,RK3E cells transfected with blank vector;③ experimental group,RK3E cells transfected with C5aR shRNA.After incubation with LPS for 12 h,the ratio of apoptosis was tested by flow cytometry,the level of mRNA was tested by RT-PCR,and binding of ~(125)I-rrC5a to RK3E cells stimulated with LPS were performed to examine the expression of C5aR in RK3E cells.Results:Compared with the normal control group and negative control group,in the experimental group the ratio of apoptosis was significantly decreased(P＜0.01),and the expression of C5aR mRNA was significantly inhibited(P＜0.01),and binding of ~(125)I-rrC5a to RK3E cells was significantly decreased also.Conclusion:Hairpin shRNA targeting C5aR gene can lead to obvious gene silence in vitro and inhibit the cell apoptosis induced by LPS in kidney epithelial cell.