Objective To investigate the mid-term outcome of high tibia osteotomy (HTO) combined with tibial tuberosity limited rotation in the treatment of disorder of patellofemoral joint with knee varus.Methods HTO was performed in all 42 patients.The lower limb alignment,lateral femoral tibial angle,medial proximal tibial angle,Q angle and TF-TG were measured before and after operation.All the patients with disorder of patellofemoral joint were divided into three types based on preoperative patellar trajectory and TF-TG.According to different types we chose the limided rotation of distal tibia through adjustment of tibial tubercle location to improve patellar track.Type Ⅰ in 10 cases and type Ⅱ in 12 cases only accepted HTO,type Ⅲ in 20 cases accepted HTO and tibial tubercle rotation.The rotation angle was determined according to preoperative imaging results and foot forward angle.Before and after operation patellar tilt angle,patellofemoral angle and lateral patellar angle were measured.Results 42 patients were followed up for more than 5 years,an average of 72.20± 12.60 months (60-88 months).No nerve damage,infection and bone nonunion occurred.The patellar tilt angle was improved from 12.45°±3.76° to 6.98°± 1.78° 5 years after operation.The patellofemoral angle was improved from 12.51°±4.71° to-4.70°±2.57° 5 years after operation,and the lateral patellar angle was improved from 1.50°±4.90° to 7.80°±3.10°.Preoperative lateral femoral tibial angle was improved from 182.45°±2.20° to postoperative 174.60° ±3.50°,medial proximal tibial angle was improved from preoperative 78.75°± 3.50° to postoperative 93.25°±1.95°.The Lysholm score was improved from preoperative 53.10±5.60 to 92.70±5.50.The preoperative IKDC subjective score was improved from 47.50±6.40 to 91.30±6.90.The Kujala score was 62.40±8.70 before operation and 87.30±4.10 5 years after operation.Visual analogue score (VAS) was 7.50±2.45 before operation and 1.50±0.90 5 years after operation.The short and mid-term follow-up,knee patellar trajectory and lower limb alignment was good,postoperative symptoms was satisfactory and acceptable.Conclusion HTO combined with limited rotation of the tibial tubercle could improve patellar trajectory and lower limb alignment to relieve pain,and has satisfactory mid-term outcome in the disorder of patellofemoral joint with knee yarus patients.

Objective To analyze and compare the clinical effects of comprehensive treatment and liver transplantation for hepatocellular carcinomas (HCC)exceeding Milan criteria.Methods A retrospective analysis was performed on the clinical data of 157 patients with HCC (exceeding Milan criteria)who received comprehensive treatment (99 patients,including 48 cases meeting Shanghai criteria)and liv-er transplantation (58 patients,including 26 cases meeting Shanghai criteria)in our hospital from January 2006 to January 2010.The medi-an survival times and 1 -,2-,and 3-year survival rates were calculated by the Kaplan-Meier method,and the log-rank test was used for survival difference analysis.Results For all patients,the median survival times of the comprehensive treatment group and liver trans-plantation group were 18.00 ±1.15 and 23.40 ±4.44 months,respectively,and their 3 -year survival rates were(18.2 ±3.9)% and (39.7 ±6.4)%,respectively;the survival was significantly improved by the treatment of liver transplantation (P=0.009).For patients meeting Shanghai criteria,the median survival times of the comprehensive treatment group and liver transplantation group were 20.00 ±1.17 and 36.00 ±0.00 months,respectively,and their 3-year survival rates were (25.0 ±6.3)% and (57.7 ±9.7)%,respectively;there was significant survival difference between the two groups (P=0.008).For patients exceeding Shanghai criteria,the median survival times of the comprehensive treatment group and liver transplantation group were 16.00 ±1.78 and 16.00 ±1.69 months,respectively,and their 3-year survival rates were (11.8 ±4.5)% and (25.0 ±7.7)%,respectively;there was no significant survival difference between the two groups (P=0.221).Conclusion Compared with comprehensive treatment,liver transplantation leads to a significant higher long-term survival rate in HCC patients exceeding Milan criteria but meeting Shanghai criteria.However,the two therapies cause no significant survival difference in HCC patients exceeding Shanghai criteria.

BACKGROUND: To avoid acute rejection,it is necessary to use imunosuppressive drug regimen for long term to control immune state.However,imunosuppressive drug regimen of allogenic organ transplantation increases infection incidence of recipients,and induction of allograft immunological tolerance might be an ideal method for solving these problems.The long-term immunologic tolerance has been able to be induced in the experimental rodent models.Among these protocols,donor bone marrow cell (DBMC) infusion exerts an important role in the induction of allograft immunological tolerance.OBJECTIVE: To investigate effects of combination of cyclosporine A (CsA) with DBMC infusion on heterotopic rat cardiac allograft survival time.DESIDN: A randomized controlled animal experiment.SETTING: Renal Disease Center,First Affiliated Hospital of Zhejing University School of Medicine.MATERIALS: This study was performed at the Laboratory Animal Center,Zhejiang University School of Medicine between March 2002 and December 2005.Inbred male Lewis rats (n=40,serving as donors) and male BN rats (n=60,serving as recipients) of SPF grade were used in this study.The protocol was approved by the Hospital's Ethic's Committee.METHODS: Forty rats prepared for heterotopic rat cardiac allograft were randomly divided into 4 groups,with 10 rats in each: control group,in which,rats received no treatment,CsA group,in which,rats received CsA infusion for 7 days successively; CsA +DBMC group,in which,rats received DBMCs during and 6 days after the surgery and additional 7 successive days of CsA infusion,and a DBMC group,in which,rats received DBMCs infusion during and 6 days after the surgery.In addition,BN rats that received beterotopic rat cardiac allograft served BN controls.The survival time of heteroropic rat cardiac allograft was investigated.Serum interleukin-2 level and tumor necrosis factor-α mRNA expression level in the transplanted cardiac allograft were measured. The percentage of antigen presenting cells (APC) from donor,CD3+CD25+ cells,CD4+CD25+ cells,CD86+ cells,and the ratio for CD4+CD45RC+ and CD4+CD45RC- in the recipient peripheral blood karyocytes were measured by flow cytometry 6,12 and 18 days after surgery.MAIN OUTCOME MEASURES: The survival time of beteruropic rat cardiac allograft,serum interleukin-2 (IL-2)level,tumor necrosis factor- α (TNF- α ) rnRNA expression level, rejection grading,the percentage of DBMCs in the recipient peripheral blood karyocytes,CD3+CD25+ cells,and CD4+CD25+ cells,as well as CD86 expression,and the ratio for CD4+CD45RC+ and CD4+CD45RC.RESULTS: Forty Lewis male rats and sixty male BN rats were all included in the final analysis. The heterotopic rat cardiac allograft survival time was longer in the CsA +DBMC group than in the control group and DBMC group (P ＜ 0.05). Serum IL-2 level and TNF- α mRNA expression were respectively lower in the CsA +DBMC group than in the control group and DBMC group ( P ＜ 0.05).The rejection was milder in the CsA +DBMC group than in the remaining 3 transplantation groups.In the CsA +DBMC group,CD 86 expression in the recipient peripheral blood karyocytes was markedly inhibited,and 6 and 12 days after surgery,the ratio for CD4+CD45RC+ and CD4+CD45RC- and the percentage of CD3+CD25+ were respectively lower compared to control group and DBMC group.DBMCs in the recipient peripheral blood karyocytes were more in rats that received DBMC infusion compared to rats that received no BDMC infusion.CONCLUSION: Short-term CsA treatment combined with DBMC infusion can lower acute rejection of heterotopic rat cardiac allograft and prolongssurvival time of cardiac allograft.

Through protein-protein BLAST of homologous sequences in different species in NCBI database and preliminary simulating molecular docking and molecular dynamics by computer software discovery studio 3.1, three amino acids R25K26K27 of natural human parathyroid hormone (1-34) with Q25E26L27 were mutated and the biological activity of the mutant peptide was evaluated. Result showed that: root mean superposition deviation RMSD value between PTH (1-34)-(RKK-QEL) and PTH (1-34) peptide main chain was 2.509 3, indicating that the differences between the two main chain structural conformation was relatively small; the interaction energy between PTH (1-34)-(RKK-QEL) and its receptor protein PTH1R had been enhanced by 7.5% compared to nature PTH (1-34), from -554.083 kcal x mol(-1) to -599.253 kcal x mol(-1); the number of hydrogen bonds was increased from 32 to 38; PTH (1-34)-(RKK-QEL) can significantly stimulate the RANKL gene expression (P < 0.01) while inhibiting the OPG gene expression (P < 0.01) in UAMS-32P cells; in the co-culture system of UAMS-32P cells and mouse primary femur bone marrow cells, PTH (1-34)-(RKK-QEL) stimulated the formation of osteoclasts (P < 0.01) and had a higher biological activity than PTH (1-34) standard reagents.

Objective To investigate the analgesic effect of salmon calcitonin(sCT)and its effect on expression of calcitonin receptor(CT-R)in periaqueductal gray(PAG). Methods Rat models of neuropathic pain were prepared by chronic constriction injury(CCI). Thermal withdrawal latency(TWL)and mechanical nociceptive threshold(MNT)were measured using hot plate test and yon Frey monofilaments test. The distribution of CT-R in PAG was detected by immunohistochemical method. CT-R protein was quantitatively determined by western blotting. Fourty male SD rats were randomized into 5 groups: normal group, sham-CCI group, CCI group, CCI plussubcutaneous sCT group, and CCI plus microinjection of sCT into PAG group. Results TWL, MNT, andexpression of CT-R in PAG showed no difference between normal group and sham-CCI group(P＞0. 05). TWL and MNT in CCI group were significantly lower than those in normal group(P＜0.05), and expression of CT-R in CCI group was significantly higher than that in normal group(P＜0.05). TWL, MNT and expression of CT-R in CCI rats increased significantly after sCT therapy(P＜0. 05), and the effect was more marked in PAG injection group than subcutaneous injection group(P＜0.05). Conclusions sCT raises the pain threshold and increase the expression of CT-R in PAG of CCI rats, while PAG injection showed more marked effect than subcutaneous injection.

Objective To evaluate the MR perfusion imaging in measuring the hemodynamics of liver.MethodsCT and MR perfusion imaging were performed in livers of 13 pigs.The hepatic artery perfusion(HAP),portal vein perfusion(PVP),total hepatic blood flow(THBF),portal vein perfusion index(PVI),distribution volume(DV)and mean transit time(MTT)were calculated and compared respectively.Hemodynamics of the liver from two kinds of imaging was compared by paked t-test,and the relativity was analyzed by Pearson correlation analysis.ResultsIn CT and MR perfusion imaging of the 13 pigs:the hepatic artery perfusion(HAP)were(37.7±7.38)and(35.80±7.31)ml-min-1·100ml-1.the portal vein perfusion(PVP)were(123.16 35.89)and(121.40±36.81)ml·min-1·100 ml-1.the total hepatic blood flow(THBF)were(160.88±37.05)and(157.21±38.71)ml·min-1·100 ml-1.the portal vein perfusion index(PVI)were(75.95±5.21)%and(76.63±5.24)%,the distilbution volume(DV)were(35.10±11.17)%and(41.03±10.06)%。The mean transit time(MTT)were(14.08±1.40)and(14.94±1.32)s.There were no statistical differences for HAP,PVP,THBF and PVI between CT and MR perfusion imaging(t=1.263,0.926,1.225,-1.115,P＞0.05).There were no statistical differences for DV and MTT(t=-3.345,-3.200,P＜0.01).Analysis of these two kinds of imaging showed the correlation coefficient of PVP,THBF and PVI were hishly related(r＞0.85,P＜0.01).Correlation coefficient of HAP,DV and M1T were moderately related(r＞0.70,P＜0.01).ConclusionsMR perfusion Call quantify the blood flow parameters of liver accurately.It is close to that of the CT perfusion imaging.

Objective To explore the effects of movement on hippocampal β-amyloid protein ( Aβ ) and amyloid precursor protein (APP) in senescence-accelerated and senescence-prone (SAMP8) mice, and the mechanism by which movement improves learning and memory in mice with a model of Alzheimer's disease. Methods Forty 3-month-old SAMP8 mice were divided randomly into a movement group and a control group. The movement group was trained with a running wheel 10 min daily, 5 days a week in the first month, and 20 min daily in the second month. Morphological changes in the hippocampus were observed under the microscope after HE staining. The expression of Aβ in the hippocampus was detected by immumohistochemical methods and APP mRNA expression was detected by RT-PCR two months later. Results HE staining showed neuron degeneration and death, chromatin condensation and vacuolar degeneration in the hippocampus of the 5-mouth-old SAMP8 mice of the control group. The movement group showed less neuron degeneration and death, and the morphology of most cells was normal The expression of Aβ in the hippocampus of the 5-month-old SAMP8 mice in the movement group was significantly lower than that in the control group. APP mRNA expression levels in the movement group were also significantly lower.Conclusions Movement can delay neuron degeneration and down-regulate Aβ and APP mRNA expression levels in the hippocampus of SAMP8 mice. It may be an important mechanism by which movement improves learning and memory in mice with a model of Alzheimer's disease.

Objective:To investigate the clinical value of Octreotide plus Ulinastatin in the treatment of severe acute pancreatitis(SAP)in elderly patients.Methods:From May 2016 to February 2019, 124 elderly SAP patients admitted to the gastroenterology department of our hospital were enrolled and divided into the combination therapy group and the control group, with 62 patients in each group.The combination therapy group was treated with Octreotide and Ulinastatin, while the control group was treated with Octreotide alone.Serum leukocyte count, C-reactive protein(CRP), interleukin(IL)-6, tumor necrosis factor(TNF)-α and amylase levels were monitored before and 1, 3, 5, 7 and 14 days after treatment by automated biochemical analysis and enzyme-linked immunosorbent assays.The pain grade scale, APACHE-Ⅱ score and efficacy evaluation were analyzed for the two groups 7 days after treatment.The time to oral refeeding and length of hospitalization were compared between the two groups, and related complications during the treatment were recorded.Complications and the recurrence of pancreatitis within 1 year were followed up in both groups.Results:There was no statistically significant difference in serum white blood cell count, CRP, amylase, IL-6 and TNF-α levels between the two groups before treatment(all P>0.05). Serum white blood cell count, CRP and TNF-α levels had significant differences( t=3.735, 2.851 and -2.147, P=0.036, 0.029 and 0.043)and serum amylase and IL-6 levels had no significant difference( P>0.05)between the two groups 3 days after treatment.All the above parameters had significant differences between the two groups 7 days after treatment( t=3.624, 2.918, -2.166, 2.684 and -2.593, P=0.023, 0.011, <0.001, 0.015 and <0.001). Serum amylase, IL-6 and TNF-α levels had significant differences( t=-3.515, 4.627 and -3.189, all P<0.001)and serum white blood cell count and CRP had no significant difference(all P>0.05)between the two groups 14 days after treatment.There were significant differences in visual analogue scale(VAS)and APACHE-Ⅱ score between the two groups 7 days after treatment( t=-2.346 and -3.245, P=0.021 and 0.002). On the 7th day after treatment, the effectiveness rate was 79.0%(49/62)in the combination therapy group and 61.3%(38/62)in the control group, with a significant difference between the two groups( χ2=4.661, P=0.031). Compared with the control group, time to oral refeeding and hospitalization length were shorter in the combination therapy group than in the control group(6.72±1.87 d vs.7.65±1.69 d, 11.23±2.98 d vs.13.85±3.42 d, t=-2.868 and -4.565, both P<0.05). There were significant differences in the incidences of infectious pancreatic necrosis, gastrointestinal adverse reactions and organ failure between the combination therapy group and the control group(11.3% or 7/62 vs.25.8% or 16/62, 43.5% or 27/62 vs.21.0% or 13/62, 1.6% or 1/62 vs.11.3% or 7/62, χ2=4.324, 7.233 and 4.810, P=0.038, 0.007 and 0.028). There were significant differences in mean length of time without complications and recurrence between the combined group and the control group(10.25±3.26 months vs.8.72±3.73 months, 10.69±2.51 months vs.9.62±2.92 months, Log Rank χ2=7.463 and 4.589, P=0.006 and 0.032). Conclusions:Octreotide combined with Ulinastatin can effectively alleviate local symptoms, slow clinical progression, reduce the risk of complications, decrease the recurrence rate and promote early recovery in elderly SAP patients.

AIM: To construct a recombinant lentivirus RNAi vector carrying cytochrome C oxidase gene to obtain the titer of the lentiviral stock for investigation of the expression in the eukaryotic cell and the affection of the COX gene silencing in the eukaryotic cells. METHODS: According to the DNA of the cytochrome C oxidase gene, we designed and synthesized complementary single-strand DNA oligos, annealed the single-stranded oligos to generate a ds oligo, cloned the ds oligo into pENTR/U6 to obtain an entry clone; An LR recombination reaction was performed between the pENTR/U6 entry construct and pLenti6/BLOCK-iT-Dest to generate expression construct, the 293FT cell line was cotransfected with pLenti6/BLOCK-iT expression construct, and the viral packaging mix, viral supernatant was harvested to determine the titer. RESULTS: The DNA sequence of interest clone to the vector was constructd to generate an entry clone and an expression clone successfully, which were proved by sequence determination. A vector producing cell line 293FT was established, and the titer for transfection was obtained. Western blotting analysis demonstrated that COX shRNA expression construction could suppress the expression of MTCOX-I. CONCLUSION: A lentivirus RNAi vector containing cytochrome C oxidase gene was successfully constructed.

Objective To analysis the malignant performance characteristics of tumor stem cell-like side popula-tion cells in patients with cervical cancer. Methods The cervical cancer cells were obtained from surgical resection tumor tissue. The tumor stem cell-like side population cells were isolated by flow cytometry. The cell growth curve was drawn by MTT assay. The invasion ability of tumor cells was compared by transwell assay. The clonogenic capacity was detected by clone formation in soft agar. The expression level of ABCG2 protein, a drug-resistant gene, was detected by immunofluores-cence method. Finally, these cells were transplated into the subcutaneous of de thymus mice. The rate of tumor formation was compared between groups. Results The results from flow cytometry assay showed the percentage of cervical cancer stem cell-like side population cells was 1.39%. Compared with the non-side population cells, the side population cells grow quickly, showed the enhanced invasion ability and colony forming ability. There was more high expression level in ABCG 2 protein of side population cells. The tumor form rate was 100%(10/10) in the side population cells and the non-side popula-tion cells was 20%(2/10). Conclusion The cervical cancer stem cell-like side population cells have more malignant perfor-mance characteristics than that of non-side population cells, which maybe a core target for cancer gene therapy in the future.