Objective:To construct mammalian cell expression vector of human leukocyte antigen C gene and express it in JAR cell and study its functions.Methods:Total cell RNA was extracted from peripheral blood mononuclear cells and the cDNA was amplified by RT PCR;the cDNA fragments were directly inserted into Pbluescript KS(+/-) and the recombinant PBS HLA C was identified by restriction endonuclease digestion and sequencing;then the plasmid of pcDNA HLA C was transfected into JAR cells.Lactate Dehydrogenase(LDH) release assay was employed to detect the cytotoxicities of NK cells proliferated in vitro against HLA C transfectants JAR cells.Results:The recombinant mammalian cell expression vectors of pcDNA3 HLA C was constructed,the sequences of the insert was identical to the published sequences encoding HLA Cw *0602 gene.Several transfected JAR clones were obtained,which expressed HLA Cw *0602 in high level and constitutively.The specific lysis was detected for NK cells stimulated by IL 2 against JAR cells and HLA C transfectants JAR cells.But both cells were resistant to freshly isolated PBL cells.Conclusion:The recombinant mammalian cell expression vectors of pcDNA3 HLA Cw *0602 was successfully constructed,JAR cells resistance to NK lysis could involve an HLA C independent mechanism.

Objective:To explore the influence of chaperone tapasin on HLA E expression on cell surface.Methods:The infection method was used to transfer the recombinant retrovirus expression vector which was constructed with HLA E molecules on target cells was detected with FACS technique.Results:Exogenous HLA E expression on T2 cell surface was detected with 74.13% and no HLA E expression was detected on control cells and that was transfected with empty vector.Conclusion:It is possible that the expression of HLA E molecules on cell surface involves in TAP independent mechanism.

Objective:To investigate the mechanism of association between Pregnancy induced Hypertension and HLA G.Methods:The distribution and frequencies of HLA G alleles were analyzed in 54 PIH patients and 146 random healthy controls with nonradioactive digoxigenin labeled PCR SSO method.Results:The results showed that the same nine HLA G alleles were detected and the G*01011 was the commonest allele with the frequency of 31.38% and 37.33% respectively in both PIH and controls groups,the next was G*01013,25.00% and 20.21% separately.Statistic analysis showed that the frequencies of HLA G alleles were similar to in the two groups and there was no significant difference between PIH patients and controls.Conclusion:These data suggest that there is no significant detectable relation between the HLA G polymorphism and PIH.

Objective:To investigate the effects of recombinant human interluekin-1 receptor antogonist (rhIL-1Ra) on the cartilage repair in rat temporomandibular joint(TMJ) with osteoarthritis(OA).Methods:Collagenase-Ⅱ was injected into bilateral TMJs of 24 adult rats for the induction of bilateral TMJOA,1 week after injection,5μg rhIL-1Ra(diluted in 0.05 ml normal saline) was injected into each right TMJ and the left joint received the same amount of normal saline injection as the control.12 animals were sacrificed at 2and 4 weeks after the first injection respectively.HE staining,immunnohistochemical method and RT-PCR examination were conducted.Mankins scere was used to evaluate the TMJOA degree.1 adult SD rat was used as healthy control,and sacrificed at 2 weeks of the experiment.Results:The TMJs of both sides showed typical OA-related cartilage degradation 2 week after IL-1Ra treatment,the Mankin~ score of the IL-1Ra treated and control joints was 1.33±0.52 and 2.00±6.63 (P＞0.05),4 week after treatment that was 3.00± 0.63 and 6.50 ± 0.84 (P＜0.05),respectively.Lower expression of ADAMTS-4 and ADAMTS-5 was observed in the treated joints than in the controls (P＜0.05).Conclusion:Intra-articular injection of IL-1Ra into TMJ can alleviate the cartilage lesion,the mechanism may lie in the inhibition of the expression of ADAMTS-4 and ADAMTS-5.

Objective To investigate serum nesfatin-1 levels in type 2 diabetes (T2DM)patients with or without nonalcohol-ic fatty liver(NAFLD).Methods The hight,weight,serum GLU,lipid profiles,INS,HbA1c,ALT,nesfatin-1 levels were measured in 21 1 T2DM patients[98 without NAFLD(TD2M without NAFLD group)and 1 13 with NAFLD(TD2M with NAFLD group)] and 75 normal subjects(NGT group).BMI、HOMA-IR were also calculated.Results The serum nesfatin-1 level was significantly higher in T2DM group than those in NGT group(P <0.05),and T2DM with NAFLD group had higher nesfatin-1 level than T2DM without NAFLD group (P < 0.05 ).There was a close relation between nefastin-1 of T2DM group and course of disease (r=-0.447,P <0.05).NAFLD existed a significant correlation with BMI、TG and nesfatin-1 (P <0.05).Conclusion nesfatin-1 may partially contribute to the pathogenesis of T2DM and NAFLD.

Objective To evaluate the clinical features of osteoarticular damage of patients with brucellosis,and to enhance the knowledge of clinical doctors.Methods The demographic characteristics,clinical presentations,accessory examination of 265 patients with brucellosis admitted from January 2011 to April 2014 in Ji'nan Infectious Diseases Hospital were retrospectively analyzed.Patients were classified into two groups:Patients with osteoarticular complications and patients without osteoarticular complications.Results Of the 265 patients,forty-six patients (17.36％) had osteoarticular complications.Spondylitis was the most common complication [50.00％ (23/46)],followed by sacroiliitis [41.30％ (19/46)] and peripheral arthritis [26.09％ (12/46)].Sacroiliitis was more common than other types of osteoarticular damage in females [21.74％ (10/46)],whereas in males spondylitis was more common [36.96％ (17/46)].Peripheral arthritis was more common in individuals under the age of 17 years old [4.35％ (2/46)],and sacroiliitis were more common in the ages of 18-60 years old group [23.91％(11/46)] and spondylitis were more common in over 60-year age group [30.43％ (14/46)].Patients with osteoarticular complications showed longer duration of illness from the onset of the symptoms to its diagnosis (median 45 days),compared to those non-osteoarticular brucellosis (median 30 days,Z =5.74,P ＜ 0.05).Conclusion Due to diversity of osteoarticular damage in patients with brucellosis,in case of a patient with long-term fever and osteoarticular symptoms,medical and epidemiological history should be inquired so as to strengthen early diagnosis and treatment of the disease based on combination with radiological findings.

Objective To investigate the antitumor mechanism of astragaloside combine with cetuximab on the colon cancer cell line RKO of EGFR over expression through cell proliferation and autophagy. Methods The cell proliferation of colon cancer cell line RKO intervened by astragaloside with or without cetuximab was detected by 4- methyl- teerazolium (MTT). The expressive changes for protein of EGFR 、P62 and LC3 in RKO cells was detected by western bolt. Results MTT showed that 100 ug/mL astragaloside combined with 120 ug/mL cetuximab had significantly inhibiting effect on RKO cells of EGFR expression. Western blot showed that astragaloside can affect the expression of P62 and LC3 to suppress the occurrence ofautophagy. Conclusion In the vitro studies showed that astragalosidecan enhance the antitumor cell proliferation of cetuximab through inhibit autophagy in the treatment of colon cancer.

[Abstrct] Objective The deformity of severe varus knee (varus angulation > 15°) can be treated by many ways in primary total knee arthroplasty , but the efficacy and cost are different. The topic of this study provides a new surgery: autograft (wedge grafts) implant and substitute medial tibial bone defects. Methods From Jan. 2006 to Jan. 2014 , a retrospective study was conducted in the First Affiliated Hospital of BaoTou Mendical College. Out of 687 patients, 721 primary knees were operated. Bone defects in proximal tibia (varus angulation >15° ) were encounted in 16 patients in 21 knees in the pattern of non-inclusive slope bone defects in the mean varus degree of 27°. Their IBM was 31 kg/m2. The average age of patients was (63 ± 5) years. The score of clinical knee society score (KSS) before operation was 18 on average. Among of them, the pain score were 12 points, the average range of motion (ROM) were 12 points. the average funtional score were 20 points. During operation,the slope bone defect of the medial tibial plateau was tailored into wedge-shaped ladder bone defect by osteotomy,and then the defect was filled with the resected tibial plateau autograft in which the thickness and shape were matched completely. The high-intensity cortical part of the autograft was placed in the rim to sustain the tibial prosthesis. Three months after operation ,patients started to walk without the help of crutch. Results The average followup period was 5.3 y , We observed that an average postoperative scores of clinical KSS from 42 points to 88 points with the score of functional KSS from 57 points to 92 points. Fusion was observed by trabeculae crossing from the graft to the tibial metaphysis on plain films at least 6 mouths without collapse, stress fracture, untight and bone ununion. Confirmation of vascularization of the graft was observed in bone scans of nuclide. showing increased uptake in the operated area. Inflammatory makers were normal. Conclusion Autograft (wedge grafts) reconstruction of medial tibial bone defects, is of high rate of healing in implant bones, good restoration of the function and low cost.

Genetic Diseases, Inborn ; economics ; prevention & control ; therapy ; Humans ; Metabolic Diseases ; economics ; prevention & control ; therapy

ObjectiveTo determine optimal positive end expiratory pressure (PEEP) in mechanical ventilation in children with severe asthmatoid disease based on the quasistatic pressure-volume (P-V) curve.MethodsA serf-control study was done on 23 children with severe asthmatoid disease in the pediatric intensive care unit( PICU ).Quasistatic lung P-V curve of these patients was analyzed and the lower inflection point (LIP) from P-V curve was determined.Three different PEEP (0 cm H2O,LIP,LIP+2 cm H2O,1 mm H2O =0.098 kPa) were given to the patients.The effects of PEEP at different levels on gas exchange,hemodynamic and airway pressure were observed.ResultsThe quasistatic LIP were (2.70 ±2.00)cm H2O.When PEEP was increased to the level of LIP + 2 cm H2O,PaO2 / FiO2 and lung compliance improved significantly (P ＜ 0.01 ) and dynamic lung compliance was the highest,peak inspiratory pressure was (22.30 ± 3.00) cm H2O and mean airway pressure was( 14.11 ± 1.01 ) cm H2O,without obvious adverse effects on mean arterial blood pressure and heart rate.There was no difference in PaCO2,when compared PEEP =0 cmH2O to PEEP =LIP + 2 cmH2O.ConclusionThe application of PEEP is safe.LIP + 2 cm H2O from quasistatic P-V curve could be set as the optimal PEEP under which mechanical ventilation has the best efficacy and do not aggravate CO2 retention and abnormality of hemodynamics in children with severe asthmatoid disease.