Objective:To construct the eukaryotic vector of human DC-SIGN and EGFP fusion protein,and to identify the protein in cell line COS7.Methods:RT-PCR,T-vector and pEGFP-C1 vector were used to construct the recombinant expressing plasmid encoding for the fusion protein of DC-SIGN and EGFP.COS7 cells were transfected with the plasmid.Real-time PCR and laser scanning confocal microscope were used to quantificate expression of the fusion protein and cell function of uptaking BCG.Results:DC-SIGN cDNA was successfully cloned into the eukaryotic vector pEGFP-C1.The recombinant vector was transfected into COS7,real-time PCR test showed the amount of mRNA encoding for the fusion protein was 4.52?1011 copies/ml and laser scanning confocal microscope confirmed that the cells could uptake BCG.Conclusion:We have constructed a recombinant vector expressing DC-SIGN and EGFP fusion protein.

Objective To study the preoperative treatment and prognosis observation? in sensitized recipients of kidney transplantation.Method Forty-one recipients positive for preoperative PRA accepted renal allograft transplantation from January 2007 to July 2012.All recipients were given immunosuppressant or immune induction by anti-CD25rnAb in advance,and plasma exchange,immunoadsorption and intravenous high-dose immune globulin were administered.Meanwhile,donor HLA antigens had to avoid all stored HLA antibodies of the recipient,and lymphocyte cytotoxicity cross test (CDC) had to be negative.Anti-human lymphocyte globulin (ATG) was used to strengthen the immune induction,and tacrolimus + mycophenolate mofetil (MMF) + corticosteroids triple immunosuppressive regimen was adopted after transplantation.Then intravenous micafungin would be given after transplanted kidney function was normal,and ganciclovir and sulfamethoxazole were taken orally to prevent infection.Result In 41 recipients positive for preoperative PRA,13 cases were positive for only HLA class Ⅰ antibodies,15 cases for only HLA class Ⅱ antibodies,and there existed 13 cases of both HLA class Ⅰ and class Ⅱ antibodies also with PRA≥50％.Fifteen patients achieved normal serum creatinine in one week,and no hyperacute rejection and accelerated rejection occurred.Fourteen recipients experienced an episode of acute rejection (34.15％,14/41): 12 recovered by steroids bolus therapy,and the other two reversed in 3-5 days by cyclophosphamide or ATG treatment.One case died of mycotic pneumonia in 4 months later.One-year recipient/kidney survival rate was 97.6％ (40/41).Conclusion The recipients positive for preoperative PRA only can accept renal allograft transplant while the donor's HLA antigens had to avoid all stored HLA antibodies of recipients themselves and CDC test was negative.After that the combination of desensitization therapy,immune induction therapy and postoperative potent immunosuppressant can prevent acute rejection effectively and increase postoperative recipient/kidney survival rate.

Objective To investigate the correlation between human leukocyte antigens-A,-B,-DRB1 (HLA-A,-B,-DRB1) high resolution alleles and chronic renal failure (CRF) caused by immunoglobulin-a nephropathy (IgAN).Method The polymerase chain reaction-sequence-based typing (PCR-SBT) method was used to investigate the genotypes of HLA-A,-B and-DRB1 high-resolution alleles in 191 cases of CRF caused by IgAN (experimental group) and 503 healthy blood donors (control group).The alleles frequencies between two groups were compared and the association between CRF caused by IgAN and the polyrnorphism of HLA was analyzed.Result (1) There were 25 alleles at A locus,48 alleles at B locus and 32 alleles at DRB1 locus in experimental group.(2) The genetic frequency of HLAA * 2901 [Pc =0.033,OR =10.738,95％ CI (1.193,96.691)],HLA DRB1 * 1106 [Pc =0.0001,OR =0.969,95％ CI (0.944,0.994)],HLA-DRB1 * 1202[Pc =0.002,OR =1.859,95％ CI (1.259,2.745)],HLA-DRB1 * 1401 [Pc =0.021,OR =0.984,95％ CI (0.967,0.998)],HLA-DRB1 * 1602[Pc=0.015,OR=1.915,95％ CI (1.157,3.17)] in experimental group was higher than in control group (P＜0.05).Conclusion There is susceptibility association of HLA-A * 2901,HLA-DRB1 * 1106,HLA-DRE * 1202,HLA-DRB1 * 1401,HLA-DRB1 * 1602 with CRF caused by IgAN.It is concluded that there is a close genetic and immunological correlation between HLA alleles and the pathogenesis of CRF caused by IgAN.

Objective To discuss the characteristics of in-patients with Beh(c)et's disease in Xinjiang Uygur Autonomous region.Methods Clinical data of 146 cases with Behcet's disease admitted to Xinjiang Uygur Autonomous region people's hospital were analyzed.The demographic data and clinical manifestations were analyzed retrospectively.The categorical data was compared using x2 test or Fisher's exact test.Results ① Of the 146 patients with BD,99 cases were Han,47 cases were uyghurs.Oral ulcer were observed in 146 cases (100％).Genital ulcer occurred in 115 cases (78.8％),of which 76 were Han (76.8％),39 (82.9％) were uyghurs,the difference was not significant (P=0.391).The initial presentation was oral ulcer in 83 cases (56.8％),of which 60 cases (60.6％) were Han,23 cases (48.9％) were the uyghurs.The second most common presentation was oral ulcer and genital ulcer,which occurred in 42 cases (28.8％),of which 23 cases (23.2％) were Han,19 cases (40.4％) were the uyghurs.More Han patients initially presented with both oral and genital ulcers than uyghur (P=0.032).② Skin lesions were recorded in 66 cases (45.2％),of which 45 cases (45.4％) were Han,21 cases (44.7％) were the uyghurs,the difference was not significant (P=0.930).③ Joint damage occurred in 40 cases (27.4％),of which 22 cases (22.2％) were Han,18 cases (38.3％) were the uyghurs.Joint damage wasmore frequently observed in Han than in uyghur (P=0.042).④ Eye lesion happened in 21 cases (14.4％),ofwhich 9 cases (9.1％) were Han,12 cases (25.5％) were the uyghurs,ie,eye lesion of Uyghur was higher than Han (P=0.008).⑤ Vascular lesions were detected in 8 cases (5.5％),of which the 5 cases were Han,3 cases were uyghurs,the difference was not significant (P=0.712).⑥ Nervous lesions were observed in 2 cases (1.37％),1 was Han and one was uyghur,the difference was not significant (P=0.542).Conclusion The most common clinical presentations of BD in Xinjiang region are oral and genitalulcers.Skin,joints,eye,vascular,nervous system can also be affected.Joint damage in Han is higher than Uyghur,but eye lesion of Uyghur is more common than Han.

Objective To investigate the effects of different doses of gabapentin on streptozotocin (STZ)-induced diabetic neuropathic pain in rats.Methods Male SD rats aged 6 weeks weighing 180-200 g were used in this study. Diabetes ntellitus ( DM) was induced by intraperitoneal STZ 60 mg/kg and confirmed one week later by blood glucose =16.7 mmol/L before breakfast. The DM rats were randomly divided into 4 groups ( n = 6 each) : gabapentin groups received intraperitoneal gabapentin 30, 60 and 120 mg/kg twice a day (at 9:00 am and 3:00 pm) for 3 weeks respectively and control group received intraperitoneal normal saline 0.6 ml instead of gabapentin. The paw withdrawal threshold to von Frey filament stimulation was measured before and at 30, 60, 120, 180, 240 min after first gabapentin injection and once a week for 3 weeks. Results After gabapentin 60 and 120 mg/kg, the paw withdrawal threshold to mechanical stimuli was significantly increased and lasted for about 4 h. The analgesic effect peaked at 60 min after IP gabapentin injection. Normal saline and gabapentin 30 mg/kg had no significant analgesic effect. The degree of analgesia was significantly decreased at day 14 and 21 of treatment with gabapentin 60 and 120 mg/kg as compared with that at 60 min after gabapentin injection. Conclusion The hyperalgesia and allodynia in rats with diabetes mellitus can be effectively reversed by gabapentin 60 and 120 mg/kg,while long-term use of gabapentin can induce drug tolerance.

Objective To explore the intervention effect of active fraction of Angelica Sinensis Radix in mice under high altitude hypoxia condition. Methods Totally 72 healthy SPF mice were randomly divided into control group (K), model group (M), Rhodiola rosea group, and active fraction of Angelica Sinensis Radix groups (B, C, X). The mice were administerted corresponding treatment by gavage for 21 days. Control mice were given normal saline in same volume. From the 8th day, all mice excepted control mice were exposed to high altitude hypoxia cabin after 0.5 hour gavage treament. On the 22nd day, after got out of the cabin and their body weight were measured, mice were put to death through eyeball blood sampling to prepare splenic lymphocyte suspension. The proliferation and transformation capacities of lymphocyte cell and killing activity of NK cells were detected by MTT. The content of IL-2 in the serum of mice in each group were detected by ELISA. Results Compared with the control group, the body weight of mice, the proliferation and transformation capacities of lymphocyte cell, the killing activity of NK cells, and the content of IL-2 were all significantly decreased (P<0.05, P<0.01). Experiment tests showed that the proliferation and transformation abilities of lymphocyte cell and the killing activity of NK cells were all increased in the mice of group B, C, and X compared with those of the model group (P<0.05, P<0.01). The stimulate index of lymphocyte cell was raised after X intervention compared with those of the model group (P<0.05). The content of IL-2 in the serum was enhanced after intervention of active fraction C and X of Angelica Sinensis Radix compared with those of the model group (P<0.05, P<0.01). Conclusion Active fraction of Angelica Sinensis Radix shows increasing immunological function of mice exposed to hypoxia.

Objective To explore the feasibility of coelocentesis in prenatal diagnosis.Methods Coelocentesis were applied on 58 women scheduled for at 6～12 weeks of gestation,and fetal heart rate before the procedure and 1,5,30 minutes afterwards and hemorrhage of amnionic cyst was observed.Y gene of sex-determining region was detected by polymerase chain reaction,and compared with chorionic villi sample.Results The total achievement ratio of coelocentesis was 96.6% of all cases in five minutes.The coelomic fluid was successfully aspirated at first attempt in 93.2% at 7～10 week's gestation,25.0% at 6～7weeks and 50.0% at 10 ~ 12 weeks.There was no significant difference in FHR between before coelocentesis and afterwards (P＞0.05).The detection accurate rate of SRY was 91.1%,all female fetus were matched with those observation by chorionic villi sampling,and only 5 male fetus were not matched,the accurate rate was 80.8%.Conclusion Coclocentesis has the advantage of simple procedure,safe and high feasibility.The optimal time of coclocentesis is in the 7th to 10th week of gestation.

Objective To investigate the production path of major histocompatibility complex class Ⅰ chain-related gene A(MICA) antibodies and the impact on the therapeutic efficacy after acute rejection in renal transplantation recipients.Methods Luminex flow cytometry was used to detect antiMICA antibodies and the antibody specificity in 157 pre-transplant kidney transplant recipients randomly selected.The clinical data were collected,anti-MICA antibody production pathway and immunoglobulin types were analyzed,and the impact of IgM anti-MICA antibody and IgM&IgG complex anti-MICA antibodies on acute rejection (AR) incidence and therapeutic efficacy after renal transplantation.Results Of the total 157 recipients,19 recipients were positive for anti-MICA antibodies before renal transplantation in 68 recipients who had history of blood transfusion,pregnancy and transplant sensitized experience (27.9％ ); In 89 recipients having no sensitized experience,MICA antibodies were positive in 26 recipients (29.2％ ) (P＞0.05).In 45 anti-MICA antibody-positive recipients,the anti-MICA antibodies type was IgM in 26 cases having no sensitized experience; and that was IgG and IgM complex in 19 cases having sensitized experience.In 38 antiMICA antibody-positive recipients undergoing kidney transplantation,7 out of 22 IgM anti-MICA antibodies recipients had AR (31.8％) that was reversed by methylprednisolone pulse therapy,and 7out of 16 IgM&IgG complex anti-MICA antibodies recipients had AR (43.8％) and treated with methylprednisolone pulse therapy:reversion in 3 recipients (42.9％),and the graft function loss in 4 recipients.The AR incidence was not associated with the two immunoglobulin types of MICA antibodies(P＞0.05),but there was significant difference in the reversal rate of AR (P＜0.05).Conclusion For non-allergenic history recipients,there exists the classic “natural antibodies” pathway in the production of the anti-MICA antibodies whose immunoglobulin type was IgM.In addition,the reversal effect of AR in recipients with IgM anti-MICA antibodies was much better.We need to attach importance to IgM&IgG complex anti-MICA antibodies for the pre-transplant anti-MICA antibodies in renal transplant recipients,because their AR treatment outcome is poor.

Objective To explore the effects of perinatal infection on retinopathy of prematurity (ROP).Methods A retrospective cohort study was performed to analyzed the clinical data of 238 preterm infants at gestational age ≤32 weeks who were delivered in Guangdong Women and Children Hospital from November 2014 to October 2015 and ROP screening examinations.Observation was not terminated until they were 45 weeks of corrected gestational age.Mild ROP was defined as having stage 1 or stage 2 ROP in zone Ⅱ or Ⅲ without additional disease,and severe ROP was defined as stage 3 or higher,any ROP in zone Ⅰ,prethreshold/threshold,with additional disease,and aggressive posterior retinopathy of prematurity (AP-ROP).Medical records of eligible preterm infants were retrospectively reviewed and analyzed.Occurrences of ROP,severe ROP,and clinically significant ROP requiring surgical treatment were assessed.Results The mean gestational age of the cohort was (30.10 ± 1.34) weeks (25.29-32.00 weeks) and the mean birth weight was (1 373 ± 272) g(720 ～2 330 g).ROP was diagnosed in 76 of 238 infants (31.9％),including 39 cases with mild ROP (16.4％) and 37 cases with severe ROP (15.5％).Surgical treatment was performed on 22 infants (9.2％).In the patients with ROP,the time to develop ROP from birth was (35.16 ± 14.26) d and the mean time of its most serious stage was (44.62 ± 18.99) d.In 22 patients with ROP who required surgical treatment,the time of surgical treatment was (50.27 ± 17.24) d.In univariate analysis,maternal perinatal infection disease was found to be associated with ROP occurrence (x2 =7.891,P =0.005) and ROP progression requiring surgical treatment (x2 =4.494,P =0.034).Small gestational age,low birth weight and long-term oxygen therapy were found to be asso ciated with ROP occurrence and severe ROP (gestational age:t =-5.803,P ＜ 0.001;t =-5.290,P ＜ 0.001;t =-4.150,P ＜ 0.001;birth weight:t =-4.942,P ＜ 0.001;t =-4.058,P ＜ 0.001;t =-3.126,P =0.002;the duration of oxygen therapy:t =2.351,P =0.020;t =2.473,P =0.018).Apgar scores ≤ 7 at 1 min and 5 min were found to be associated with severe ROP (x2 =4.803,P =0.028).Neonatal sepsis and neonatal fungal infection were found to be associated with ROP occurrence (x2 =6.071,P =0.014;x2 =4.070,P =0.044).Neonatal fungal infection was also found to be associated with severe ROP (x2 =5.479,P =0.019).Multivariate regression analysis indicated that maternal perinatal infection disease was associated with an increased risk of ROP and ROP progression requiring surgical treatment (OR =2.837,P =0.023;OR =4.087,P =0.012).Maternal preeclampsia was also associated with an increased risk of ROP (OR =2.506,P =0.040).Gestational age was an important risk factor for the development of ROP.The smaller the gestational age was,the higher the rate of occurring ROP and severe ROP (OR =0.518,0.508,0.520,all P ＜ 0.001).Conclusions Both fetal and neonatal exposure to infection appear to contribute to the increase of ROP risk in the preterm infants at gestational age ≤ 32 weeks.Maternal perinatal infection disease and maternal preeclampsia were independently associated with ROP occurrence and ROP progression in the preterm infants at gestational age ≤32 weeks.