Objective To explore the immunoregulatory and lethal effects of natural killer T lymphocytes(NKTs) in vitro .Meth‐ods The mixed lymphocyte cultured(MLC) system was established ,in which the B16F10‐luc‐G5 cells were set as target cells ,the total lymphocyte cells were set as effector cells .(1)In the experiment on immunoregulatory effects ,NKT lymphocytes or CD4+CD25+ T lymphocytes were set as regulating cells ,there was three groups ,including the NKT group ,CD4+CD25+ T group and pure target cell control group .Otherewise ,the 1640 blank control group was set by only adding RPMI1640 solution .(2)In the ex‐periment on antitumor effects ,the NKT or natural killer(NK) lymphocytes were set as killer cells ,there was three groups ,inclu‐ding the NKT group ,NK group and pure target cell control group .Mixed culturing 24 ,48 and 72 hours ,the bioluminescence of target cells in MCL system was detected by using the in vivo imaging system .Results (1)In the experiment on immunoregulatory effects ,there were statistically significant differences in measured average photon numbers between NKT group ,CD4+ CD25+ T group and the two control groups(P<0 .05) .The statistically significant differences were also found in the NKT group between 24 hours and 72 hours (P<0 .05) .(2)In the experiment on antitumor effects ,there were statistically significant differences in meas‐ured average photon numbers ,when the NKT group and NK group were compared to the pure target cell control group(P<0 .05) . After culturing 24 and 72 hours ,statistically significant differences were found between NKT group and NK group(P<0 .05) .Con‐clusion The NKT cells could inhibit the lethal effects of lymphocyte cells on target cells ,and the inhibitory effects are changed by the length of culturing .Compared with the CD4+CD25+ T lymphocytes ,NKT lymphocytes have strongger regulatory effects .Addi‐tionally ,the NKT cells have lethal effects on target cells ,which might be weaker than that of NK cells .

Objective To investigate the risk factors for delayed graft function (DGF) recovery in the recipients of kidney transplantation after the operation. Methods The data of 209 recipients who received kidney transplantation during January 2005 to March 2007 were analyzed retrospectively. The DGF group comprised of 47 cases. 162 cases without DGF constituted the control group. Chi-square analysis method was employed to look for the risk factors of DGF, including warm ischemia time ≥10min, hypovolaemia in the recipient, acute tubular necrosis (ATN), nephrotoxicity of calmodulin inhibitors, acute rejection (AR), 12h ≤ cold ischemia time ≤ 24h, sex, age ≥ 50, the modus of dialysis, the number of times of transplantation, human leucocyte antigen (HLA) zygosity, urinary fistula or obstruction of ureter during early period after operation, blood-transfusion before transplantation and hepatitis virus infection. Then Logistic regression analysis was used to assess and rank the relative risk of potential variables. Results The incidence of DGF was 22.5%(47/209). All patients recovered within 20.8?10.3 days except 2 cases due to AR and 1 case due to graft phlebothrombosis. The relative risk factors include AR, ATN, nephrotoxicity of calmodulin inhibitors, recipient hypovolaemia and warm ischaemia time ≥ 10min. Conclusion The ranking of risk factors is as following, ATN, nephrotoxicity of calmodulin inhibitors, recipient hypovolaemia, AR and warm ischaemia time ≥ 10min, according to decrement of the coefficient correlation. It is very important to comprehend the risk factors thoroughly in order to prevent them effectively.

Objective To analyze the clinical data of living related donor kidney transplantation,and to evaluate the related problems including the donor selection and safety,the surgical approach of living donor nephrectomy,preoperative intervention,and the relation of type matching with transplantation result.Methods Data of 30 cases of living donor kidney transplantation during 07/2002 to 07/2007 were studied retrospectively.29 cases were primary transplantation and only in one patient the transplantation was repeated,2 patients secured their transplants from the spouse,while in 28 cases transplants were donated by relatives.The blood type of all the patients was identical with an exception in 2 cases,in whom the blood type was compatible.The HLA matching results were:1 case with no-mismatch,3 cases with 4-loci mismatch,13 cases with 2-loci mismatch and 13 cases with 1-locus mismatch.The kidney was obtained with open nephrectomy in 21 cases,and in 9 donors the was harvested laparoscopically.In the 30 cases,one right kidney was donated,and the left kidneys were donated in other 29 cases.Triple-combined immunosuppressive protocols consisted of calcineurin inhibitors(CNI),MMF/Aza and steroid.Results Except in one case the allograft was removed for hyperacute rejection,and in one recipient who was again retransplanted with a cadaveric kidney due to dysfunction of transplanted kidney,28 recipients recovered smoothly in 6.3?8.0 days.In 4 cases infection with cytomegalovirus after the surgery occurred,but recovered after anti-virus treatment with ganciclovir.One patient suffered from urinary leakage,and recovered after adequate drainage,nutritional support and infection prevention.Two patients suffered from acute rejection in the first month after transplantation and reversed after hormone stosstheraphy.All donors recovered.The renal function also recovered.Conclusion Both the strict selection of donor and the overall preoperative evaluation of the donor and recipient are critical for the success of transplantation.Living relative donor kidney transplantation is worth of popalarizing,as it has preferable HLA match,short waiting time and renal ischemia time,and the source of donor kidney is expanded.

ObjectiveTo analyze the homology of Citrobacter freundii strains and detect multidrug resistant genes.MethodsThe minimum inhibition concentration was tested with standard agar dilution method and the the homology of C.freundii strains was measured with pulsed field gel electrophoresis (PFGE) and the PCR was used to perform the drug resistant genes such as β-lactamase,disappearing of outer membrane channel protein.ResultsThe four strains ( No.1,2,4,5 ) showed nineteen banding patterns with PFGE.PCR experiment showed that there were 2 strains (No.1,4) with blaCTX-M,3 strains (No.2,3,5 ) with blaDHA,1 strains(No.3) with blaACT/MIR,and 4 strains(No.1,2,4,5) with blaKPC-2.PCR analysis comfirmed that No.2 and No.4 disappearing of OmpF and No.3 disappearing both of OmpC and OmpF.The expression levels of the chromosomal ampC gene of No.3 isolates was 106.7 times higher than the negative isolates.ConclusionThere is high homology in carbapenem-resistant isolates and the mechanism of drug resistance is complex including blaKPC,AmpC,ESBLs,disappearing of outer membrane channel protein.The spreading of drug resistance result from above genes.

Objective To investigate the clinical distribution and the drug resistance of carbapenemresistant Proteus mirabilis strains isolated from 2009 to 2012 ; and to study carbapenemase genotypes of the isolates.Methods A total of 15 non-repetitive carbapenem-resistant Proteus mirabilis strains were selected from 422 Proteus mirabilis strains isolated from Ningbo First Hospital during January 2009 to December 2012.The minimal inhibitory concentrations (MIC) of 6 antibacterial agents,including imipenem (IPM),meropenem (MEM),ertapenem (ETP),ciprofloxacin (CIP),amikacin (AK) and minocycline (MIN),against 15 isolates were determined by E-test.The modified Hodge test (MHT) was used to detect the carbapenemase production in isolates.The PCR assay was performed to detect drug resistance genes of blaKPC,blaNDM-1,blaGES,blaSME,blaIMI-1/NmcA and blaSHV-38.Plasmids were extracted from the blaKPC-positive strains and then transformed into Escherichia coli J53 strains by electroporation.The transformed and untransformed Escherichia coli J53 strains were tested for MIC values and blaKPC gene by E-test and PCR respectively.Results The resistance rates of the 15 carbapenem-resistant Proteus mirabilis strains to IPM,MEM,ETP,CIP,AK and MIN were 100％,100％,100％,86.7％ (13/15),33.3％ (5/15) and 80％ (12/15),respectively.7 out of 15 strains were Hodge test positive,and 11 strains were blaKPC-2 positive.Results of PCR amplification showed that the transformed Escherichia coli J53 strains,whose MIC values to IPM,MEM,and ETP were increased by 2 to 64 times,were blaKPC-2 gene positive.Conclusion The carbapenem-resistant Proteus mirabilis strains in this study were resistant to many commonly used antibiotics,however,the resistance rates to AK were relatively low.The dominant carbapenemase genotype was blaKPC-2 carried by the plasmid.Attention should be paid to its easily transmissible feature among the strains in clinic.

BACKGROUND:Several studies have attempted to apply mouse nerve growth factor to local lesions of peripheral nerve and found that local injection of mouse nerve growth factor can promote nerve recovery, which is superior to systematic application. OBJECTIVE:To evaluate the clinical effects of gelatin sponge infiltrated by mouse nerve growth factor in the repair of peripheral nerve injury. METHODS:Thirty-six patients with single peripheral nerve injury, including 16 males and 20 females, aged 18-48 years, were randomly divided into two groups: 18 patients in case group underwent debridement and neuroanastomosis, and then the injured nerve was wrapped by gelatin sponge which was infiltrated by mouse nerve growth factor and folowed by plaster fixation, anti-inflammatory therapy, neurotrophy and circulation improvement therapy; the other 18 patients in control group were treated only with debridement and neuroanastomosis and other conventional therapies. At 4 weeks after treatment, electrophysiological examination was performed. In addition, sensory and motor function of the distal end of injured nerve was evaluated at 6 months after treatment. RESULTS AND CONCLUSION: Sensory evoked potential and motor evoked potential showed that the recovery rate was 78% (n=14) and 83% (n=15) respectively in the case group, while 57% (n=10) and 66% (n=12) in the control group. There was significant difference between the two groups (P < 0.05). The total effective rate was 94.4% (n=17) in the case group and 83.3% (n=15) in the control group, which were statisticaly better in the case group than the control group (P < 0.05). These findings indicate that it is significantly effective to treat peripheral nerve injury by gelatin sponge infiltrated by mouse nerve growth factor that has good biocompatibility.

BACKGROUND:Currently, there are large numbers of studies related to the association between colagen type I alpha1 (COL1A1) Sp1 polymorphism and bone mineral density and fracture risk, but the results are inconsistent. OBJECTIVE:To evaluate the impact of the COL1A1 Sp1 polymorphism on bone mineral density and fracture by using the Meta-analysis. METHODS:We comprehensively searched the eligible studies for the present meta-analysis through MEDLINE, PubMed, EMBASE databases. Pooled odds ratios and 95% confidence intervals of Sp1 polymorphisms for bone mineral density and fracture risk were obtained, with attention to study quality and publication bias. RESULTS AND CONCLUSION:A total of 32 studies met the inclusion criteria, among which, 22 studies evaluated the Sp1 polymorphism and fracture risk. Significant associations were found in five genetic models. In the stratified analysis by region, the same results were found in the Europeans but not Americans and Asians. Thirteen studies evaluated the Sp1 polymorphism and low bone mineral density risk. A similar result was obtained. However, the analysis of bone mineral density data showed an increased relation between Sp1 polymorphism and low bone mineral density in Europeans and Americans but not in Asians. Overal, the current meta-analysis concludes that the COL1A1 Sp1 polymorphism is associated with low bone mineral density and fracture risk, especialy in Europeans. However, susceptibility to them varies markedly among populations from different regions.

Objective To investigate the value of procalcitonin(PCT),hypersensitive C-reactive protein (Hs-CRP),D-Dimer(DD)in the diagnosis and prognosis judgment of pulmonary embolism(PE). Methods A total of 201 patients were enrolled in the study since the PE diagnosis were made in hospital from JAN. 2013 to DEC. 2016,in which they were divided into high-risk group(n=52),intermediate-risk group(n=88),low-risk group (n = 61),cure-patient group (n = 67) and 36 normal individuals were selected respectively as a blank control group. Results The average of the PCT,Hs-CRP and DD level in the high,intermediate and low-risk group were significant difference comparing with the cure-patient and control group(P<0.05). Conclusions Teh concertration of PCT and Hs-CRP were associated with different risk stratification of PE. Combined with DD , the level of PCT and Hs-CRP can be used to evaluate the risk stratification of PE and help to judge prognosis and guide treatment.

Objective To study the relationship of soluble LAIR (sCD305 and CD3060) expression in recipient serum with cytomegalovirus (CMV) pneumonitis after renal transplantation. Methods Nineteen serum specimens from recipients were divided into CMV pneumonitis group (n=10) and control group (n=9). Then the concentrations of sCD305 and CD3060 were quantitated with sandwich ELISA. The data were analyzed by using student t test. Results sCD305 was skewness distributed in both 2 groups, was 0.000-3.039 μg/L in CMV pneumonitis group and 0.000-8.375 μg/L in con-trol group. CD3060 was skewness distributed in CMV pneumonitis group and the concentration was 0.000-0.017μg/L. CD3060 was mormally distributed in control group and the concentration was 0.046±0.035 μg/L. There was significant difference of CD3060 (P=0.000) concentrations and no sig-nificant difference of sCD305(P=0.316) concentrations in 2 groups, respectively. Conclusions The concentration of CD3060 is low in CMV pneumonitis patients. The combination of CMV PP65 antigen detection and CD3060 detection is helpful for the early and precise diagnosis of CMV pneumonitis in renal transplantation patients.

OBJECTIVE To study characteristics of changing T lymphocytes in epidemic hemorrhagic fever(EHF) patients during acute phase and find out the pathogenesis,in order to elevate the level of early diagnosis.METHODS The anticoagulant blood from 30 cases of EHF patients and 50 normal healthy blood donors was collected.T lymphocyte subsets were detected by flow cytometry.RESULTS Compared with those of normal persons,CD4+ T cell counts of EHF patients decreased,CD8+T cell and double CD4+CD8+ cell(double positive cells,DP cell) counts of EHF patients increased obviously,and 25 cases of EHF in recovery stage returned to normal.And in comparison with HIV,CMV and EBV patients,DP cell counts of EHF patients increased obviously.CONCLUSIONS T lymphocytes of EHF decrease obviously but could be resumed,detection of amounts of lymphocyte subsets and CD4+CD8+ cells can provide an early diagnosis method to EHF.