Objective: To construct and express the eukaryotic expression vector of human pSecTag2B-CD226(PTA1).Methods: The gene fragment encoding extracellular region of human CD226 was cloned into the eukaryotic expression vector pSecTag2B.After sequencing,the vector was transfected into COS-7 cells,and the expressed molecule was purified by affinity chromatography.Finally,the product was characterized by ELISA.Results: hCD226-6His was successfully expressed.After purification,the concentration of hCD226-6His was 50?g/ml.Conclusion: Human CD226-6His fusion protein involving the extracellular region of CD226cDNA has been successfully expressed and purified,which helps prepare the ground for further functional studies of this molecule.

Objective To investigate the expressions of interleukin-1β(IL-1β), interleukin-6(IL-6) and tumor necrosis factor alpha(TNF-α) in cartilage of children with Kashin-Beck disease(KBD) in order to provide a possible mechanism of the disease. Methods Articular cartilage tissues of 5 KBD children(KBD group) were selected from KBD children autopsy samples keeping in Institute of Endemic Diseases, Medical School of Xi’an Jiaotong University; articular cartilage tissues of 5 normal children ( control group ) were selected from non-KBD areas of Shaanxi Province, three cases were from accident death children, two cases were the samples of congential malformation of six finger. Expressions of IL-1β, IL-6 and TNF-α in the cartilage were detected using immunohistochemistry; the cells of articular cartilage were divided into three areas (superficial zone, middle zone and deep zone) to analyze the expressions of IL-1β, IL-6 and TNF-α. Results The expressions of IL-1β in superficial zone , middle zone and deep zone of articular cartilage of KBD group (63.50 ± 7.19, 54.75 ± 5.50, 66.20 ± 9.91) were significantly higher than those of control group(5.75 ± 1.26, 0.00 ± 0.00, 0.00 ± 0.00, all P<0.05). The expression of IL-6 in superficial zone of articular cartilage in KBD group(55.25 ± 6.24) was significantly higher than that of control group(0.00 ± 0.00, P<0.05). The expressions of TNF-αin all zone of articular cartilage of KBD group(33.25 ± 6.50, 3.75 ± 0.96, 29.80 ± 1.92) were significantly higher than those of control group (3.74 ± 0.82, 0.00 ± 0.00, 0.00 ± 0.00, all P < 0.05). Conclusion The levels of IL-1β, IL-6 and TNF-α are up-regulated in articular cartilage of KBD children, suggesting that cytokines may play an important role in matrix degradation in KBD children cartilage.

Objective To investigate the role of soluble CD305(sCD305)in renal allograft rejection.Methods Concentration of serum sCD305 was detected on 20 healthy volunteers and 153 cases of recipients after kidney transplantation by using double monoclonal antibody sandwich enzyme linked immunosorbent assay.Results In the healthy volunteers and 98 recipients with normal renal funetion,sCD305 was detected at low levels of(4.3±2.3)μg/L and(6.3±3.7)μg/L.In 20 cases of acute rejection and 5 cases of graft loss,serum sCD305 levels were(36.3±14.7)μg/L and(28.8±9.4)μg/L,and significantly higher than those in the healthy volunteers and recipients with normal renal function.Meanwhile,in the 30 cases of chronic rejection and 6 cases under dialysis treatment,the levels of sCD205were(13.1±5.5)ttg/L and(11.2±4.6)μg/L and significantly higher than those in the healthy volunteers and recipients with normal renal function.Conclusions CD305 was presented at high level in the recipients with renal acute or chronic rejection,and it might be a potential marker for monitoring graft rejection after transplantation.

Objective Based on detection of the soluble LAIR-1 (sLAIR-1) and sIL-2R in the bile from recipient after liver transplant, the role of sLAIR-1 and sIL-2R in graft acute rejection were analyzed. Methods Bile sLAIR-1 level and sIL-2R were determined by double mAb sandwich enzyme linked immunosorbent assay in 55 cases of liver transplantation. Results In 22 recipients with normal graft function, sLAIR-1 and sIL-2R were detected at low level in the bile. In the 29 cases of liver acute rejection (AR), significant increase of bile sIL-2R level was detected on the lst and 2nd d before final diagnosis. With the effective methylprednisolone pulse therapy, sIL-2R level was decreased significantly on the 3rd d. On the other hand, remarkable increase of bile sLAIR-1 was found on the lst,2nd and 3rd d before final diagnosis. After of methylprednisolone pulse therapy for 3 d, bile sLAIR-1resturned to the control level. Conclusion Both bile sIL-2R and sLAIR-1 are detected at high level in the recipients suffering from liver acute rejection. The level of bile sLAIR-1 changes dramatically and responsively according to liver acute rejection. Therefore, detecting these two markers synergistically may be a promising monitor for rejection after liver transplantation.

Objective To investigate the expression of ligands of DNAM-1 and NKG2D in the colonic cancer.Methods The colonic cancer tissue and adjacent normal colonic tissues were collected from 42 colonic cancer patients who were admitted to the Tangdu Hospital of Fourth Military Medical University from June 2010 to January 2011 were retrospectively analyzed.The expressions of CD155,CD112 and MICA/B in the colonic cancer tissues and the normal colonic tissues were detected by immunohistochemistry.The expressions of CD155,CD112 and MICA/B in the colonic cell line SWll6,SW480,SW620 and Colo205 in the Duke's A,B,C and D phases were detected by cell cytometry.The relationship of the expressions of the 3 ligands and the clinicopathological parameters was analyzed using the Mann-Whitney U test,chi-square test and Fisher exact probobility.Results Week expression of CD155 was found in the normal colonic tissues,while the expressions of CD112 and MICA/B were not found.In the colonic cancer tissues,the expressions of CD155,CD112 and MICA/B were 81.0％,52.4％ and 47.6％,which were significantly increased.The expressions of CD155,CD112 and MICA/B were not correlated with the gender,tumor differentiation,lymph node metastasis and Duke's staging (P ＞ 0.05).The overall expression rates of CD155,CD112 and MICA/B in the colonic cancer cell line SWll6,SW480,SW620 and Colo205 were 88.9％,67.4％ and 42.3％,respectively.The overall expression of CD155 was significantly higher than CD112 and MICA/B (F =23.17,P ＜ 0.05).Conclusion CD155,CD112 and MICA/B express in the colonic cancer tissues and colonic cancer cell line SW116,SW480,SW620 and Colo205,and the expression of CD155 is the highest.

AIM: To investigate the effects and mechanisms of irbesartan,one of the angiotensin Ⅱreceptor blockers,on kidney function in diabetic rats. METHODS: Forty adult male Wistar rats were randomly divided into four groups: control group,diabetes group,irbesartan group and captopril group. At the end of 12 weeks,the rats were sacrificed. Urine volume,body weight,kidney weight/body weight,plasma,glucose,glycosylated hemoglobin (HbA_1c),urinary ?_2-microglobulin (?_2-MG) excretion,urinary albumin excretion rate (UAR),creatinine clearance (Ccr) were measured. Nitric oxide (NO) and endothelin-1 (ET-1) levels in plasma,urinary and renal tissues were determined. RESULTS: Urine volume,kidney weight/body weight,plasma glucose,HbA1C,UAR,Ccr,urinary ?_2-MG excretion,NO and ET-1 levels of urinary,blood and renal tissue in diabetic rats were significantly higher than those of normal controls ( P

Objectives:To determine the expressions of decoy receptors (DcR1 and DcR2) of TRAIL in carcinoma of endometrium. Methods:The expressions of DcR1 and DcR2 in endometrium tissues from 13 carcinoma of endometrium and 7 normal endometrium were detected by immunohistochemical staining.Results: The expressions of DcR1 and DcR2 in carcinoma of endometrium were much lower than in normal endometrium. Conclusions:The decreasing of DcR1 and DcR2 in carcinoma of endometrium may be concerned with its pathogenesis, which may be related to the prevention of endometrium from carcinomatous change.

BACKGROUND:Cytotoxic T lymphocyte-associated antigen 4 is a newly discovered costimulatory molecule. It has been studied more in tumor and autoimmune diseases, less in the field of kidney transplantation.
OBJECTIVE:To explore the role of cytotoxic T lymphocyte-associated antigen 4 in acute rejection after renal transplantation.
METHODS:Fifty patients undergoing renal transplantation were divided into acute rejection group (20 cases) and stable graft function group (30 cases). Another 30 healthy persons served as control group. Blood samples were extracted from the peripheral blood. Cytotoxic T lymphocyte-associated antigen 4 was detected by enzyme linked immunosorbent assay and flow cytometry.
RESULTS AND CONCLUSION:The expression of cytotoxic T lymphocyte-associated antigen 4 in the serum showed significant differences in the acute rejection group, stable graft function group and healthy control group (F=70.008 1, P=0.000 0), but showed no difference in peripheral blood lymphocytes of three groups (F=1.865 6, P=0.161 7). Compared with the healthy control group, the expression levels of cytotoxic T lymphocyte-associated antigen 4 in peripheral blood lymphocytes of acute rejection group and stable graft function group were significantly decreased (P=0.000 0). In addition, the acute rejection group had a lower cytotoxic T lymphocyte-associated antigen 4 expression than the stable graft function group (P=0.000 0). In renal transplant rejection, the expression of cytotoxic T lymphocyte-associated antigen 4 in serum was reduced, showing some correlation with acute rejection after renal transplnatation. Cytotoxic T lymphocyte-associated antigen 4 might be involved in the rejection.

Objective:To observe the value of tracer methodology combined case-based study (CBS) model in the standardized residency training of medical record writing.Methods:A total of 91 residents who were newly recruited from The Sixth Affiliated Hospital of Sun Yat-sen University in July 2019 were selected as the subjects and randomly divided into test group ( n=47) and control group ( n=44). The tracer methodology combined CBS model was used for test group while the traditional model was used for control group to compare the knowledge of medical record writing, performance of medical record writing and feedback to teaching in two groups. SPSS 21.0 was used for t test and chi-square test. Results:The correct response rate in knowledge of medical record writing, the normative scores and integrity scores after training were significantly higher than those before training ( P<0.05). The normative and integrity scores were significantly increased after training, and those in the test group were significantly higher than those in the control group [(90.3±3.5) and (91.3±3.2) vs. (88.6±3.5) and (89.8±3.0), P<0.05, respectively]. The test group was significantly superior to the control group in the autonomous learning ability [97.8% (44/45) vs. 82.9% (34/41)], communication ability [95.6% (43/45) vs. 78.0% (32/41)] and acceptance of teaching form [97.8% (44/45) vs. 82.9% (34/41)] ( P<0.05). Conclusion:The tracer methodology combined CBS model is a good way in the pre-job training of medical record writing for residents, which can mobilize the subjective initiative of the trainees and teachers, complete the teaching tasks and obtain better teaching value.

OBJECTIVETo prepare purified and concentrated coltivirus high titer antigen in order to further detect antibodies against coltivirus in serum sample of patients.

METHODSThe coltivirus in C6/36 cells was cultured and harvested at different time, and the titer was titrated. The virus was purified and concentrated by polyethylene glycol (PEG), and stored at -20 degrees and 4 degrees, with and without glycerol, respectively, then the titer of coltivirus antigen was tested by indirect ELISA. By using the antigen, coltivirus antibodies in serum samples from both suspected Japanese encephalitis (JE) and viral encephalitis (VE) patients were detected.

RESULTSThe highest titer of coltivirus was found at 3-4 weeks of culturing. The antigen titer could be maintained at least for 6 months, especially antigen with glycerol either at 4 degrees or at -30 degrees even for two years. Totally 1141 serum samples from patients diagnosed clinically as JE and VE were tested. The results showed that 130 samples were coltivirus IgM antibody positive and the average positive rate was 11.4% (130/1141). Among 41 samples of paired-serum from patients in Guangzhou Children's Hospital, 9 samples were positive, the positive rate was 22.0% (9/41) in which 5 samples were diagnosed clinically as VE.

CONCLUSIONSStable and purified coltivirus antigen was obtained in order to test coltivirus antibodies as well as development of kits. Coltivirus probably can cause summer-autumn encephalitis in China.

Antibodies, Viral ; blood ; Antigens, Viral ; isolation & purification ; Cell Line ; Coltivirus ; immunology ; Cryopreservation ; methods ; Enzyme-Linked Immunosorbent Assay ; Humans ; Reoviridae Infections ; blood