Objective To investigate the effect of microcolumn gel reagent card,micro-column glass bead reagent card and classical test tube on the antibody titer of O-type pregnant women with ABO blood group.Determination of antibody potency of pregnant women with rabbit by reagent card.Methods Using microcolumn gel reagent card,micro-column glass beads reagent card,and classical test tube parallel detection to detect the IgG blood group antibody titer of O blood type pregnant women.Results There was no significant difference in the positive detection rate between the microcolumn gel reagent card and the microcolumn glass beads reagent card at 1:128 and the test tube method at 1:64 (P＞0.05).Conclusion The clinical reference value of microcolumn gel reagent card and microcolumn glass beads reagent kit for detecting antibody titer of IgG blood group in O-type pregnant women should be set to ≥ 1:128.

Objective To evaluate the perioperative blood loss and blood transfusion in liver transplantation patients.Methods Retrospectively selected and classified 119 patients with liver transplantation medical recordsaccording to the clinical diagnosis of age,gender.Analyzed (disease)patients' blood coagulation index 24 h before surgery,intraoperative and postoperative perioperative bleeding,and blood red blood cell suspension,frozen plasma,cold precipitation,coagulation factor injection lossplateletpheresis.Results There were no significant differences in the amount of blood loss and the amount of blood preparations duringthe perioperative period of liver transplantation according to the age and sex of patients (P＞0.05);According to the clinical diagnosis of different groups,the diagnostic group (primary liver cancer group,hepatitis B cirrhosis group and severe hepatitis group) patients with a single platelet transfusion dosewas similar (P＞0.05)withsuspended red blood cells (U) at 8.4+11.9,16.2+15.7,18.1+13.5,frozen plasma (U) at 8.2+7.1,18.1+15.6 and 18.2+ 17.9 respectively andcold precipitation (U) at 9.5+ 8.2,17.1 + 16.318.5 + 16.4 (P＜ 0.05);The diagnosis group surgery before and after immediate PT (s) were 15.6+3.8,24.6+4.1,APTT (s) were 44.3+5.8,84.9+9.2,TT (s) were 20.4 +4.5,40.1+6.2,and Fib(s) were 2.6+0.8 and 1.3+0.9 (P＜0.05);however,there was no significant difference in the 24 h PT,APTT,TT and Fib between the two groups after diagnosis (P＞0.05).Conclusion Ascientific,safe and reasonable selection of perioperative blood preparation for clinical diagnosis for different liver transplantation patients is critical to the success rate and prognosis of liver transplantation;the evaluation of coagulation indexes at each interval can help guiding the blood transfusion during liver transplantation.

Objective To construct and evaluate a nomogram for predicting the risk of peri-operative complications after radical retro-pubic prostatectomy (RRP) with standard classification criteria.Methods The peri-operative complications and clinicopathological data of 240 patients (50-82 years old) who had undergone RRP for prostate cancer from June 2006 to June 2011 were retrospectively collected.Gleason score:95 cases ＜ 7; 145 patients ≥ 7;Clinical staging:cT1 is 1 case (0.4％),T2a 5 cases (2.1％),T2b 7 cases (2.9％),T2c 162 cases (67.5％),T3a 26 cases (10.8％),T3b 39 cases (16.3％).The peri-operative complications (in 30 days after surgery) were classified by Clavien-Dindo Classification system (the occurrence rates are as follows:rectum injury 1.6％,wound infection 2.0％,deep venous thrombosis 1.2％,urinary leak 5.0％,lymphocele 4.5％,myocardial infarction 5.8％,second look operation 1.6％),and a logistic regression model was used to construct the nomogram.Results BMI,N staging and Blood loss more than 200ml during surgery were independent prognostic factors of RRP morbidity in multivariate logistic regression.The nomogram predicting the risk of peri-operative complications showed relative good concordance index (0.633) and good calibration. Conclusions Based on the clinicopathological factors,a nomogram to predict the probability of peri-operative complications in patients undergone RRP was constructed.This statistical tool may be beneficial in judging operation risk and help consulting with patient before or after surgery.

Objective To evaluate the recurrence patterns and possible prognostic factors in primary penile cancer treated with penectomy and standard lymphadenectomy.Methods The records of 73 patients with penile squamous cell carcinoma who underwent penile amputation and regional lymphadenectomy between 1990 and 2005 were reviewed.The median follow up was 32 ( 16 - 183) months.Molecular markers including p53 and Ki-67 were detected using immunohistochemistry.Recurrence patterns,pathological characteristics,and patient outcomes were analyzed.ResultsTwenty( 27.4％ ) patients developed recurrences at a mean of 11 (6 - 17 ) months postoperatively and 3 cases remained alive at last follow-up.The mean cancer specific survival of the recurrent patients was 26 ( 16 - 83 ) months.Distant metastasis and multiple recurrences were common among patients with high-grade tumor (P =0.017 ).Univariate analysis revealed that higher primary tumor stage,higher pathological grade,lymph node metastasis,extra-nodal involvement and positive p53 protein expression were significantly associated with decreased 3-year disease free survival rates.Cox regression identified that only pathological grade ( P =0.025 ) and lymph node status ( P =0.024 ) were independent predictors of disease free survival.ConclusionsPathological grade and lymph node status were independent predictors of disease free survival in patients with penile squamous cell carcinoma in this cohort.Patients with high-grade disease are likely to recur in a distant or multiple pattern.

Objective To investigate the effect of songling xuemaikang(SL-xmk)pretreatment on the expression of matrix metalloproteinase-9(MMP-9)after focal cerebral ischemia-reperfusion in rats.Methods A total of 45 male Sprague-Dawley rats were randomly allocated into SL-xmk pretreatment,sham operation,and normal saline control group.Preventive gavage was per-formed for 8 weeks in rats using SL-xmk(937.50 mg/kg)suspension in the SLxmk pretreatment group(n = 15);the preventive gavage was performed in rats using the equal volume of normal saline in the sham operation(n = 15)and normal saline control(n = 15)groups.At the end of the pretreatment process,a model of middle cerebral artery occlusion(MCAO)in rats was induced by suture method for 2 hours and reperfusion for 24 hours.The effects of SL-xmk pretreatment on the neurologic deficit scores after transient MCAO,brain water content,and infarct volume in rats were observed.Immunohistochemical staining was used to detect the MMP-9 immunoreactive positive cells in ischemic brain tissue.Results The neurologjc deficit scores(1.21 ± 0.25 vs.2.37 ± 0.35,P = 0.000),the brain water content (76.24% ± 7.09% vs.88.78% ± 6.57%,P = 0.000),the percentage of infarct volume (22.62% ±2.17% vs.27.84% ±3.43%,P =0.000),and the numbers of MMP-9 positive cells(16.20 ± 2.17/mm vs.20.60 ± 2.71/mm,P = 0.000)were all significantly lower than those in the control group.Conclusions SL-xmk pretreatment may significantly inhibit the expression of MMP-9 in the brain tissue of focal cerebral ischemia-reperfusion rats and reduce brain water content and infarct volume.

Objective To investigate the institution of extracorporeal membrane oxygenation (ECMO) for primary graft dysfunction (PGD) after lung transplantation (LT) and analyze its clinical outcome.Method A retrospective analysis was performed on 22 patients with grade 3 PGD in early stage after LT from September 2002 to December 2013.There were 7 patients with single LT and 15 patients with bilateral LTx.Ventilatory support was used at early stage for 6 cases,and at later stage,ECMO assistant circulation was used for 16 cases.Result Of 6 patients treated by adjusting low volume,high frequency and high positive end expiratory pressure ventilation (PEEP) mode,2 cases reversed,and 4 cases died of respiratory failure.In 16 cases accepting ECMO support,10 cases were given venous-venous mode and 6 cases venous-artery mode.The average flow time was 5.5 days.ECMO was successfully withdrawn in 10 cases and 6 cases died of multiple organ failure,infection and cardiac arrest.Conclusion The high incidence of PGD causes high mortality peri-operatively after LT.Preventing ECMO can improve the survival rate of the lung transplant patients.Once PGD happens,appropriate treatment should be given as soon as possible.ECMO can effectively promote the transplanted lung function recovery,and reduce the perioperative mortality.If the indications of ECMO use was reached,the institution of ECMO should be used as soon as possible.

Objective To study the phylogenetic evolution and genetic variations of gag gene among the prevalent human immunodeficiency virus (HIV )‐1 strains in Guangxi Zhuang Autonomous Region . Methods Plasma samples of 158 HIV‐1 infected patients in Guangxi area were collected during October 2011 to March 2012 .The gag gene fragments of HIV‐1 were amplified by reverse transcription/nested‐polymerase chain reaction and then sequenced .MEGA 5 .03 was utilized to construct phylogenetic tree and to calculate the genetic distances and selection pressures (globle ω) of gag gene and its coding regions . The comparisons between two groups were tested by Student′s t test ,and the comparisons of multiple groups were tested by one‐way ANOVA .Results A total of 140 amplification products of gag gene were obtained from 158 samples .Four subtypes of HIV‐1 were found ,including CRF01_AE (80 ,57 .1% ) , CRF08_BC (46 ,32 .9% ) ,CRF07_BC (10 ,7 .1% ) ,and subtype B (B′) (4 ,2 .9% ) .The genetic distances of gag gene of the above subtypes were 0 .036 ± 0 .001 ,0 .031 ± 0 .002 ,0 .043 ± 0 .003 and 0 .102 ± 0 .006 ,respectively ,with statistical significance (F=220 .62 ,P<0 .01) .The p17 and p24 coding regions suffered negative selection pressure (globleω<1) .Neither the globle ω in p17 region nor that in p24 region had significant differences among different subtypes (F=0 .761 ,P=0 .469 and F=0 .037 ,P=0 .964 , respectively ) . Conclusion CRF01_AE is the major subtypes of HIV‐1 in Guangxi Zhuang Autonomous Region .The coding regions of gag gene are relatively conserved during evolution .Changes of HIV‐1 prevalence ,however ,may affect the genetic variation of gag gene ,which should be continuously monitored .

Objective:To predict and identify HLA-A * 0201 restricted CD8+ CTL epitopes in Mycobacterium tuberculosis (Mtb) antigen Ag85C, so as to provide evidence for epitope-based study for tuberculosis (TB) vaccine. Methods: The online database SYFPEITHI was applied to predict the potential HLA-A * 0201 restricted epitopes from Ag85C, an antigen of Mycobacterium tuberculosis. T2 cell line was used to assay the affinity between the predicted peptides and HLA-A * 0201 molecules. The specific CTL lines were induced from peripheral blood mononuclear cells (PBMCs) of HLA-A * 0201 positive TB patients and PPD+ healthy donors by peptides with high binding affinity to HLA-A * 0201 molecules. IFN-?production, in vitro proliferation and cytotoxicity of peptide-induced CTL were determined to screen HLA-A * 0201 restricted CD8+ CTL epitopes from those candidates. Results: Fourteen potential epitopes were identified from the SYFPEITHI database. After binding affinity assay, 3 of the 14 peptides (170-178 aa, 317-325 aa, and 144-153 aa) were found to have high binding affinity to HLA-A* 0201 molecules. However, only one peptide (144-153 aa) stimulated its specific CTL to release IFN-y, proliferate in vitro and produce specific cytotoxicity. Conclusion: We have successfully identified a HLA-A * 0201 restricted CD8+ CTL epitope of Mtb Ag85C-FLTREMPAWL( 144-153 aa) , which might be a candidate epitope for TB vaccine designing. Our findings provides a basis for developing novel and effective anti-TB vaccine.

ObjectiveThe procedure of peroral endoscopic myotomy (POEM) was practiced in porcine esophagus-stomach model and the efficacy and safety of POEM for patients with achalasia were evaluated.MethodsThe ex-vivo esophagus-stomach was obtained and the pylorus was closed by hemostatic forceps.The entrance of the esophagus was fixed to foamed plastics.To perform POEM,a submucosal tunnel was created,after which the circular muscle layer was dissected and the mucosal entry was finally closed with clips.After successful animal experiment,we performed POEM on 4 patients with achalasia.Results POEM was completed in 5 porcine training models,in which 2 procedures were successful without any complication,2 were complicated with perforation in muscularis propria layer and 1 with rupture in mucosal layer.POEM was successfully performed in 4 patients with achalasia,with a mean operation time of 110min.The mean length of the submucosal tunnel was 10.5cm ( ranging 8-11 cm) and the mean length of myotomy of circular muscle layer was 7 cm (ranging 5-8 cm).The resting pressure of lower esophageal sphincter (LES) decreased from 52.4 mm Hg before POEM to 19.9 mm Hg.Massive bleeding occurred at the gastroesophageal junction in the first case and perforation occurred in mucosal layer during endoscopic hemostasis.All patients were followed up for 1-4 months,and the symptom of dysphasia was relieved significantly.ConclusionThe ex-vivo esophagus-stomach model can be used as training model for procedure of POEM,enabling endoscopists with enough experience for its use in patients.POEM is an effective therapy for achalasia,while the long-term efficacy and managements for complications are still to be elucidated.

Objective To investigate the polymorphism of human cytomegalovius UL143 gene of low passage clinical isolates in Guangzhou,China.Method PCR was performed to amplify the entire HCMV ULl43 gene region of 3 clinical isolates,which had been proven by multiplex PCR.The amplification products were cloned into pMD18-T-Vector and subjected to sequencing.The result of DNA sequences were analyzed together with the one of published homologous sequences in GenBank from 14 clinical isolates.Result There were several stop codons in UL143 gene due to a base deletion in open reading frame (ORF) of D3 isolate,which could lead to produce non-functional protein.UL143 ORF of Toledo isolate consisted of 279 nueleotides,encoding a protein with 92 amino acids.UL143 ORFs of other isolates consisted of 252 nueleotides,encoding a protein with 83 amino acids.The DNA sequences were quite conserved and all the variations were base substitution.The amino acid sequences of different isolates were highly conserved.with variation of 1.2%-2.4%.There were no additional or deleted sites of post translational modification of UL143 protein in all clinical isolates except Toledo isolate.There were some differences in the secondary structure among different isolates.The isoelectric point of UL143 protein of all clinical isolates except Toledo isolate was 8.75.Conclusion All DNA and deduced amino acid sequences of UL143 gene shared great similarity among HCMV clinical strains regardless of their polymorphism.