Objective:To construct the recombinant adenovirus of AT2R by using the method of homogenous recombination in bacteria. Methods:AT2R gene was get from the vector of PUHD-AT2R by PCR ,and subcloned into shuttle vector of pAdTrack-CMV, forming transfer vector of pAdTrack-CMV/AT2R.Then it was linearized with PmeⅠ and transformed into Adeasier-1 cell. The DNA of identified recombinant plasmid was digested with PacⅠ and transfected to 293cells to package adenovirus. The PCR technique was used to detect target gene . The titre of the Ad-AT2R was measured with the aid of GFP expression. Results:PCR test indicated each the recombinant adenovirus contained the insert of AT2R. The titre of purified recombinant adenovirus was 1.5?10 12pfu/ml. Conclusion:The method of homologous recombination in bacteria is more convenient and efficient compared with that in cell .The prepared Ad-AT2R paves a sound foundation for further study.

Objective To evaluate the impact of autoantibodies to angiotensin Ⅱ type 1 receptor AT1-AA on clinic outcomes of delayed graft function (DGF) grafts.Method We reviewed the records of all 139 consecutive adult recipients who received single kidney transplantation and clinical management between Jan.2010 and Dec.2012 in our centre.The serum levels of AT1-AA were measured by a streptavidin-enzyme-linked immunosorbent assay.All patients with DGF were enrolled in this study and divided into two groups:(1) AT+ DGF group (serum AT1-AA positive,11 cases) ;(2) AT-DGF group (serum AT1-AA negative,23 cases).All clinical and laboratory data were recorded in our transplant database system at each visit.Result 139 recipients were enrolled.The overall presence of DGF was 24.5％ (34/139).The incidence of DGF in patients with high binding AT1-AA was significantly higher than that in those with low binding of AT1-AA (11/24 vs.23/115,45.8％ vs.20.0％,P＜0.05).In addition,longer duration of renal replacement therapy (59 ± 32 vs.47 ± 26 months,P＜0.05),higher resistance index (0.80 ± 0.10 vs.0.72 ± 0.10,P＜0.05) of allografts and more severe acute tubular injury (2.7 ± 0.5 vs.1.8 ± 1.1,P＜0.05)/acute tubular necrosis (0.9 ± 0.5 vs.0.5 ± 0.3,P＜0.05) were observed in AT + DGF group than in AT-DGF group.One-year graft survival and death censored graft survival were similar between two groups (90.9％ vs.95.7％,P＞0.05).Conclusion Presence of high binding anti-AT1 receptor had detrimental impacts on initiation and development of DGF.

Objective To evaluate the validity and reliability of the Chinese version of Alcohol Withdrawal Scale (AWS). Methods Totally 175 patients diagnosed as alcohol dependence according to the criteria of ICD-10 were studied. Intraclass correlation coefficient (ICC) analysis was applied for examining interrater consistency and Cronbach' s α for internal consistency. Factor analysis was used to examine the construct validity. Correlation analysis between AWS and CGI,Revised Clinic Institute Alcohol Withdrawal Syndrome Scale(CIWA-Ar) were conducted to evaluate the criterion validity. Based on clinical criteria,ROC curve was calculated so as to test the discriminative ability and establish the cut-off point of the scale. Results ( 1 ) Reliability: ICC value was 0.93,and Cronbach's α was 0.83,which indicated good interrater and internal consistency. (2) Validity:the correlation coefficients of the two subscale with the total scale score were 0.78,0.83 respectively. The correlation coefficients between the subscale were 0. 81 and factor analysis revealed that each item of the scale had relatively high load on the primary factor (0.409 ～0.926). At the time of admission,the total score of the AWS was positively correlated with that of CGI ( r = 0.71, P < 0.05 ). The total score of the AWS also was positively correlated with that of CIWA-Ar ( r = 0. 86, P<0. 05). As treatment went on,total score of the AWS showed a downward trend,at the end of the first week,the total score of the AWS was positively correlated with that of CGI ( r = 0.62, P<0.05). (3)The cut-off point of AWS for mild alcohol withdrawal state was determined as ≥3. With this cut-off point,AWS had both high sensitivity (92.1% ) and specificity (73.5% ) ,and the area under curve (AUC) was 0. 91. The cut-off point of AWS for moderate withdrawal state was determined as ≥7, and the sensitivity and specificity of AWS were 94.3 % and 89.7 % respectively, with the AUC of 0.94. The cut-off point of AWS for severe withdrawal state was determined as ≥ 10. With this cut-off point AWS had both high sensitivity (94. 9% ) and high specificity (92.6% ) .with the AUC of 0.93. Conclusion AWS has good reliability and validity and can reflect the change of the disease and the efficacy of treatment.

To investigate the epidemic and evolutionary trends of enterovirus (EV) in the external environment of Yunnan Province, China, molecular typing was performed on 4 EV strains that were isolated from environmental sewage in Yunnan. The VP1 region of isolates was amplified by RT-PCR using universal enterovirus primers, and the amplified VP1 region was sequenced for GenBank BLAST search and genotype analysis. The 4 EV strains were identified as ECHO7. Their nucleotide and amino acid homologies with the VP1 sequences of 68 ECHO7 strains retrieved from GenBank were measured by Mega software analysis. Our findings showed that ECHO7 strains from environmental sewage and population samples were in different evolutionary branches. These strains showed typical geographical and temporal differences; In addition, there were different transmission chains at the same time and in the same area. ECHO7 strains isolated from sewage water and patients with acute flaccid paralysis during the same period in Yunnan belonged to different clusters and evolved at different speeds. Special concerns are needed for this problem. Continuous molecular biological surveillance of human EV in the external environment of Yunnan will provide strong support for early warning of EV diseases.
China ; Databases, Genetic ; Enterovirus ; genetics ; isolation & purification ; Evolution, Molecular ; Humans ; Molecular Epidemiology ; Sequence Analysis ; Sewage ; virology

OBJECTIVETo observe the correlation relationship between acupuncture at Dicang (ST 4), Hegu (LI 4) and Houxi (SI 3) on the affected side of peripheral facial paralysis patients and activated areas in brain functional areas and central regulation mechanism of acupuncture at Hegu (LI 4) treatment.

METHODSEighteen cases with left peripheral facial paralysis were randomly divided into a Hegu group, a Dicang group and a Houxi group, 6 cases in each group. They were treated with electroacupuncture at left Dicang (ST 4), Hegu (LI 4) and Houxi (SI 3), respectively, and were examined with fMRI covering the whole brain at the same time. The fMRI data was analyzed by SPM software.

RESULTSIt was found that the left precentral gyrus area and the left postcentral gyrus area were activated when electroacupuncture at left Hegu (LI 4), and the right precentral gyrus area and the bilateral postcentral gyrus area were activated when electroacupuncture at left Dicang (ST 4), and there was no activated area at precentral gyrus area and post central gyrus area when electroacupuncture at left Houxi (SI 3).

CONCLUSIONThe sensory importation information from Hegu (LI 4) and Dicang (ST 4) can converge and coincide in the brain and may influence each other.

Acupuncture Points ; Acupuncture Therapy ; Adult ; Aged ; Aged, 80 and over ; Brain ; diagnostic imaging ; physiopathology ; Facial Paralysis ; diagnostic imaging ; physiopathology ; therapy ; Female ; Human Body ; Humans ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Radiography ; Young Adult

OBJECTIVEIBS-D rat model was established to assess the effect of Shen warming Pi strengthening method (SWPSM) for intervening diarrhea-predominant irritable bowel syndrome (IBS-D) by observing rats' general state, stool properties, AWR ranking, and histopathological changes.

METHODSTotally 72 rats were randomly divided into 6 groups, i.e. the normal group, the model group, the high, middle, low dose SWPSM groups, and the control group, 12 in each group. The IBS-D rat model was successfully established referring to AL-Chaer ED's modeling method. After modeling high, middle, and low dose SWPS Recipe boil-free granules were given by gastrogavage to rats in corresponding treatment groups. Sishen Pill boil-free granule was given by gastrogavage to those in the control group. Equal volume of normal saline was given by gastrogavage to rats in the model group. The medication lasted for 2 weeks. Rats' general state, stool properties, abdominal withdrawal reflex (AWR) ranking, and histopathological changes were observed.

RESULTSAfter treatment, the general state of all rats got im- provement to various degrees. The improvement in the high and middle dose SWPS Recipe groups were superior to that in the low dose SWPS Recipe group and the control group (P < 0.05). There was no statistical difference in the growth rate between after and before treatment in each group (P > 0.05). Compared with the model group and the low dose SWPS Recipe group, the defecation amount within 4 h was less in the high and middle dose SWPS Recipe groups and the control group (P < 0.05). The Bristol ranking score, average ranking of loose stool, ratio of dry stool and wet stool were lower in the high and middle dose SWPS Recipe groups than in the control group and the low dose SWPS Recipe group (P < 0.05). The AWR ranking score was lower in the high and middle dose SWPS Recipe groups than in the control group when the volume of balloon dilation was 1.5 mL. There was no organic change of histological or morphological observation.

CONCLUSIONSHigh sensitive IBS-D model was proved to be reliable. SWPSM could reduce the quantity of stools, lower Bristol ranking score, average ranking of loose stools as well as ratios of dry stool and wet stool, contributing to reducing the high sensitivity of rats' visceral organs to some extent.

Animals ; Diarrhea ; drug therapy ; Disease Models, Animal ; Drugs, Chinese Herbal ; therapeutic use ; Irritable Bowel Syndrome ; drug therapy ; Male ; Phytotherapy ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo study the influence of the reference values for semen analysis proposed in the 5th edition of the WHO Laboratory Manual for the Examination and Processing of Human Semen on the indication spectrum for intrauterine insemination (IUI).

METHODSWe retrospectively analyzed the clinical data of 111 cycles of IUI by the reference values for semen analysis in the 4th edition of the WHO Laboratory Manual (group A) and 84 cycles by the 5th edition (group B). We recorded and compared the percentages of various indications for IUI between the two groups.

RESULTSThe complications for IUI in groups A and B were as follows: asthenospermia (87.4% [97/111] vs 55.9% [47/84], P < 0.05), oligospermia (0 vs 0), teratospermia (51.4% [57/111] vs 35.7% [30/84]) , abnormal liquefaction (0.9% [1/111] vs O) , sexual dysfunction and genital malformation (0 vs 3.6% [3/84] , immune infertility (0.9% [ 1/111] vs O), and unexplained infertility (3.6% [4/111] vs 2. 4% [2/84 ] ). There were no significant differences between the two groups in the percentages of all the indications except that of asthenospermia.

CONCLUSIONThe reference values for semen analysis proposed in the 5th edition of the WHO Laboratory Manual for the Examination and Processing of Human Semen have an evident influence on the indication spectrum for IUI by largely reducing the cases of IUI for male factors, prolonging the cycles of some patients, causing excessive diagnosis and treatment for females, and increasing their mental and economic burdens.

Adult ; Contraindications ; Female ; Humans ; Insemination, Artificial ; Male ; Pregnancy ; Reference Values ; Retrospective Studies ; Semen ; Semen Analysis ; World Health Organization

OBJECTIVEPulmonary vascular structural remodeling induced by high pulmonary blood flow is an important pathologic basis of pulmonary hypertension with congenital heart disease of left-to-right shunt. However, the mechanism is still not clear. The present study aimed to examine the alteration of endogenous nitric oxide (NO) pathway in high pulmonary blood flow-induced pulmonary vascular structural remodeling, so as to explore the role of NO pathway in pulmonary hypertension induced by high pulmonary blood flow.

METHODSSixteen male SD rats were randomly divided into control group (n = 8) and shunting group (n = 8). Aortocaval shunting was produced for 11 weeks in shunt rats. Pulmonary artery mean pressure (mPAP) of each rat was evaluated using right cardiac catheterization. The ratio of right ventricular mass to left ventricular plus septal mass [RV/(LV + S)] was detected. Pulmonary vascular micro-and ultra-structure was examined by using a light microscope and a transmitted electronic microscope. Meanwhile, the concentration of plasma NO was measured by spectrophotometry. The expressions of endothelial NO synthase (eNOS) mRNA and protein by pulmonary arteries were detected by in situ hybridization and immunohistochemistry, respectively.

RESULTSAfter 11-week aortocaval shunting, mPAP was significantly increased [(22.5 +/- 2.6) mmHg vs. (15.8 +/- 2.8) mmHg, 1 mmHg = 0.133 kPa, t = 4.97, P < 0.01], and RV/(LV + S) was also markedly increased (0.267 +/- 0.022 vs. 0.221 +/- 0.016, t = 4.85, P < 0.01). The percentage of muscularized arteries was obviously increased in shunt rats compared with controls [(23.2 +/- 2.4)% vs. (13.5 +/- 2.1)%, t = 7.82, P < 0.01], and relative medial thickness of pulmonary arteries was obviously increased in shunt rats [median pulmonary artery: (7.76 +/- 0.56)% vs. (4.82 +/- 1.03)%, t = 6.23, P < 0.01; small pulmonary artery: (11.94 +/- 0.66)% vs. (6.91 +/- 0.53)%, t = 14.96, P < 0.01]. Ultrastructural changes, such as hyperplasia and degeneration of endothelial cells, irregularity of internal elastic laminar and hypertrophy and the increased number of synthetic phenotype of smooth muscle cells, were found in intrapulmonary arteries of shunt rats. Meanwhile, plasma NO concentration was increased [(30.2 +/- 7.9) micromol/L vs (19.7 +/- 5.7) micromol/L, t = 3.05, P < 0.01) and eNOS mRNA and protein expressions by pulmonary arteries were significantly augmented in rats of shunting group.

CONCLUSIONThe upregulation of eNOS/NO might be an adaptive response of pulmonary circulation to an increased blood flow in the development of pulmonary hypertension and pulmonary vascular structural remodeling.

Animals ; Blood Flow Velocity ; Hypertension, Pulmonary ; physiopathology ; Immunohistochemistry ; In Situ Hybridization ; Male ; Nitric Oxide ; blood ; Nitric Oxide Synthase ; blood ; genetics ; Nitric Oxide Synthase Type III ; Pulmonary Artery ; physiopathology ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley

Objective To investigate the CT findings of Madelung's disease in the head and neck region,and to evaluate the value of CT in demonstrating the Madelung's disease in the head and neck region.Methods CT findings of Madelung's disease in the head and neck region in 7 cases were analyzed retrospectively.All were males,with the age from 36 to 60 years,mean 51 years.All patients were underwent CT native scan,and enhanced CT scan was performed on 3 of them.Results CT images in the neck of all patients showed accumulation of nonencapsulated fat within the subcutaneous tissue and(or) deep to the platysma,and(or)within the spaces between the muscles.The fat deposits were ill-defined and symmetrical.In most cases the fat deposits involved the anterior part of the neck(infrahyoid and suprahyoid),submandibular region,the subcutaneous tissue of the nape and deep to the stenomastoid muscles.Conclusions Madelung's disease in the head and neck region have characteristic CT findings,and CT has great value in qualitative and quantitative diagnosis in Madelung's disease.

OBJECTIVETo observe the variation and significance of natural killer T (NKT) cells in patients with severe multiple injuries.

METHODSPeripheral blood was drawn from 30 patients with severe multiple injuries and 20 healthy individuals. NKT cells and the subsets of NKT cells were stained and analyzed on fluorescence activated cell sorter (FACS) using Cellquest software. The level of IL-4 and IFN-gamma in blood serum was detected by ELISA.

RESULTSThe proportion of NKT cells was significantly increased. CD4+ NKT cells was increased (t equal to -3.11, P less than 0.01) and CD4+CD8+NKT (double negative NKT, DN NKT) cells decreased in patients with severe multiple injuries compared with healthy controls (t equal to 2.99, P less than 0.01). There was a positive correlation between the proportion of NKT cells and injury severity score (ISS) by Spearman correlation analysis (r equal to 0.70, P less than 0. 01). The level of IFN-gamma was significantly decreased and the level of IL-4 significantly increased in patients with severe multiple injuries.

CONCLUSIONSWe demonstrate that human NKT cells are increased in trauma patients. Most significantly, there is an association between ISS and NKT cells. The increased CD4+NKT cells may contribute to the reduction of Th1 cytokine production and the growth of Th2 cytokine production, leading to the suppression of immunity after injury.

Cytokines ; blood ; Humans ; Multiple Trauma ; immunology ; Natural Killer T-Cells ; immunology ; T-Lymphocyte Subsets ; immunology ; Th1 Cells ; immunology ; Th2 Cells ; immunology ; Trauma Severity Indices