A new nested-polymerase chain reaction (nested-PCR) assay was developed to detect human parvovirus B19 DNA corresponding to the nonstructural protein in clinical specimens in a routine diagnostic laboratory. The sensitivity of this highly specific assay was up to 0.005 fg of B19 DNA. Parvovirus B19 was identified in sera of 20 pregnant women with abnormal pregnant outcome. Among these 20 cases, intrauterine parvovirus infection did exist in 7 pregnant women because parvovirus B19 DNA was detected in the pregnant tissues of them such as placenta tissues, chorionic villi, amniotic fluid, fetal spleen, liver and abdominal fluids.
DNA, Viral/*analysis ; Parvoviridae Infections/*virology ; Parvovirus B19, Human/genetics ; Parvovirus B19, Human/*isolation & purification ; Placenta/virology ; Pregnancy Complications, Infectious/diagnosis ; Pregnancy Complications, Infectious/*virology ; Viral Nonstructural Proteins/*analysis

BACKGROUND: Numerous studies concerning dysferlin focus on muscle diseases (such as muscular dystrophy), but the relationship between membrane repair after exercise-induced muscle damage and dysferlin is little reported. OBJECTIVE: To observe the cell membrane permeability and expression levels of dysferlin and calpain3 in the rat gastrocnemius after acute eccentric exercise, so as to provide an theoretical reference for exploring the molecular mechanism of muscle regeneration and the exercise therapy of muscular diseases. METHODS: Thirty-two Sprague-Dawley rats were randomly divided into four groups including control, 24, 48 and 72 hours post exercise groups. The membrane permeability and expression levels of dysferlin and calpain3 were determined by immunohistochemistry, western blot assay and qRT-PCR. RESULTS AND CONCLUSION: The activity of serum creatine kinase, and expression levels of calpain3 mRNA and dysferlin protein, as well as membrane permeability at 24 hours post exercise were significantly greater than those in the control group (P ＜ 0.05 or 0.01). The expression level of dysferlin mRNA at 24 and 48 hours post exercise was significantly higher than that at 72 hours post exercise (P ＜ 0.05). Therefore, the damage to the skeletal muscle cell membrane was the most severe at 24 hours after eccentric exercise. Due to Ca2+influx, expression of calpain3 mRNA was activated, and then the damaged cell membrane was repaired by increasing the expression of dysferlin.

AIM: To evaluate the efficacy of combined therapy for amblyopia in children by making use of pattern visual evoked potential ( P-VEP) game.
METHODS: This was a prospective case control study. These asthenopic children were divided into two groups. The control group ( 66 eyes of 49 patients ): occlusive therapy with glasses, cover, precision work, red light treatment and so on, later the stereo vision training was added. The experimental group (72 eyes of 52 patients):conventional methods mentioned above with P - VEP games.
RESULTS: The total effective rate and cure rate of experimental group in 6mo were higher than those of control group. The overall effective rate was 94. 4% in the experimental group and 83. 3% in the control group. There was a statistically significant difference between them (P<0. 05).
CONCLUSION: The comprehensive therapy by making use of P-VEP game is an individualized effective new way in treating amblyopia.

Objective To study the effects of exogenous phosphocreatine (CP) on brain injury after cardiopulmonary resuscitation (CPR) in rats.Methods A total of 160 male adult SD rats were randomly ( random number) divided into 4 groups:sham-operation control group ( group A),CPR group ( group B),low-dose CP group ( group C),high-dose CP group ( group D),and each group was further divided into 5 subgroups (n =8) as per study at different intervals,0.5,3,6,12 and 24 h after restoration of spontaneous circulation (ROSC) in groups B,C and D or after tracheotomy in group A.Cardiac arrest (CA) was induced by using asphyxia to establish CPR model in group B,C and D.The CP0.5 g/kg used in group C or CP 1.0 g/kg used in group D was injected into femoral vein at beginning of ROSC.Rats in each subgroup were sacrificed and the tissues of frontal lobe of brain of rats were taken at different intervals.The levels of adenosine triphosphate (ATP),adenosine diphosphate (ADP) and adenosine monophosphate (AMP) in cerebral cortex were measured by high performance liquid chromatography (HPLC),and values of total adenine nucleotides ( TAN ) and energy charge ( EC ) were detected.The activities of Na+ -K+ -ATPase and Ca2+ -Mg2+ -ATPase in cerebral cortex were assayed by spectrophotometric method. The pathological changes of cerebral cortex were observed under optical microscope.The experimental data were processed with analysis of variance by using SPSS 16.0 package. Results Compared with group A,the levels of ATP,TAN,EC,Na + -K + -ATPase,Ca2+ -Mg2+ -ATPase were lower ( P ＜ 0.05 or P ＜ 0.01 ) at each interval in groups B and C,and at intervals of 0.5,3,6,12 h in group D,and the levels of AMP were higher (P ＜ 0.01 ) at each interval in group B and at intervals of 0.5 h and 3 h in groups C and D.Compared with group B,the levels of ATP,TAN,EC,Na + -K + -ATPase,Ca2 + -Mg2+ -ATPase were higher ( P ＜ 0.05 or P ＜ 0.01 ),and the levels of AMP were lower ( P ＜ 0.05 or P ＜ 0.01 ) at intervals of 6,12 and 24 h in groups C and D.Compared with group C,the levels of ATP at interval of 24 h and TAN,Na +- K + -ATPase,Ca2 + -Mg2 + -ATPase at intervals of 6,12 and 24 h were higher in group D ( P ＜ 0.05 or P ＜ 0.01 ).There were severe pathological changes in cerebral cortex in group B,and mild changes in groups C and D. Conclusions There was obvious energy metabolism disorder after CPR in rats.Treatment with exogenous CP could increase the levels of ATP and activities of ATPase,alleviate pathological changes,especially in high-dose,and mitigate injury in cerebral cortex after CPR in rats.

Objective To analyze variations of hepatic vein in healthy people with 64-slice spiral CT.Methods Seventy-five healthy subjects underwent multi-slice spiral computed(MSCT)hepatic venography.The anatomy of the junction of the hepatic veins with the inferior vena cava and the intrahepatic drainage territory of the hepatic veins and tributaries were evaluated.The hepatic veins were classified according to three anatomic classification(Nakamura's,Marcos's and Kawasaki's classification)methods respectively.Results There was a common trunk of the middle and left hepatic veins before joining the IVC in 86.7%(65/75)of the cases.In 13.3%(10/75)of the cases,the three main hepatic veins joined the IVC separately.The ratios of Nakamura's classification type A,B,C of hepatic veins were 49.4% (37/75),37.3%(28/75),and 13.3%(10/75)respectively.The ratios of Marcos's classification type A,B,C of hepatic veins were 56.0%(42/75),24.0%(18/75),and 20.0%(15/75)respectively. The ratios of Kawasaki's classification type Ⅰ,Ⅱ of hepatic vein were 40.0%(30/75)and 60.0% (45/75).Conclusion Multi-slice spiral CT hepatic venography can provide visualization of peripheral hepatic venous branches in details.

Objective To evaluate the homogeneity of the proficiency test samples to verify whether it meets the requirements of the comparison in international reference laboratory.Methods According to the Guidance on Evaluating the Homogeneity and Stability of Samples Used for Proficiency Testing (CNAS-GL03),14 biochemical indexes including ALT,AST,ALP,GGT,CK,LDH,TP,T-Bil,Urea,Cr,UA,Glu,TG and TC in the past three years (from 2014 to 2016)were tested by the Roche detection system Modular P800 Biochemical analyzer.The mean ((x)),standard deviation (s)and coefficient of variation (CV)of the samples were calculated.One-way analysis of variance (ANOVA)was performed and the guideline of Ss ≤0.3σ was used to evaluate the between-bottle differences.Results The results showed that the CVs of AST in RELA 2014A and B were higher than 2.0％.The CVs of CK were over 2％ in all tests except for RELA 2016B.The results of ANOVA for RELA samples demonstrated that the F value of CK was over the critical value 4.39,which was statistically significant (P ＜ 0.05).The F values of the ALT and T-Bil in 2015B and the Cr in 2014A were also over 4.39 (P ＜ 0.05) respectively,while the F values of other measurements were less than the critical value of F,indicating there was no statistical significance (P ＞ 0.05).The CK measurement data Ss ＞ 0.3σ in all the samples by the guideline of Ss ≤ 0.3σ,suggesting that there was a between-bottle difference in CK.The other indexes were Ss ≤ 0.3σ,showing no between-bottle difference in those items.Conclusion There were significant differences between the bottles of the CK item in the past three years,and the homogeneity of all the other items in the samples could meet the requirements of Proficiency Testing for the international reference laboratory.

A new nested-polymerase chain reaction (nested-PCR) assay was developed to detect human parvovirus B19 DNA corresponding to the nonstructural protein in clinical specimens in a routine diagnostic laboratory. The sensitivity of this highly specific assay was up to 0. 005 fg of B19 DNA. Parvovirus B19 was identified in sera of 20 pregnant women with abnormal pregnant outcome. Among these 20 cases, intrauterine parvovirus infection did exist in 7 pregnant women because parvovirus B19 DNA was detected in the pregnant tissues of them such as placenta tissues,chorionic villi, amniotic fluid, fetal spleen, liver and abdominal fluids.

OBJECTIVETo evaluate the outcome of allogeneic hematopoietic stem cell transplantation (allo-HSCT) from HLA-matched sibling donor (MSD allo-HSCT) for severe aplastic anemia (SAA).

METHODSThe clinical data of 41 SAA patients received MSD allo-HSCT from May. 2003 to Aug. 2011 were analyzed retrospectively. 24 patients were male, 17 were female. Median age was 23 (5 - 43) years old. 28 patients had SAA-I, 9 had SAA-II, and 4 had post-hepatitis aplastic anemia. 17 patients received allogeneic bone marrow (BM) transplantation (allo-BMT), and 24 received allogeneic peripheral blood stem cell (PBSC) transplantation (allo-PBSCT). The conditioning regimens: 20 patients received cyclophosphamide (CY) + anti-thymocyte globulin (ATG) + fludarabine (Flu), 21 received CY + ATG + Flu+ cytarabine (Ara-C) ± busulfan (Bu)/melphalan (Mel). Prophylaxis for graft-versus-host disease (GVHD): 25 patients received cyclosporine (CSA) plus short-term methotrexate (MTX), 16 received tacrolimus (FK506) plus short-term MTX. The median number of infused CD34(+) cells were 3.48 (2.39 - 4.80)×10(6)/kg in allo-BMT and 2.95 (1.27 - 5.98)×10(6)/kg in allo-PBSCT, respectively.

RESULTSHematopoietic reconstitution was observed in all 41 patients (100%). The median time of neutrophils (ANC) reached to 0.5×10(9)/L and platelets (PLT) reached to 20×10(9)/L were 14 (10 - 23) days and 19 (8 - 38) days, respectively. 12 patients developed acute GVHD (aGVHD), out of which 11 developed grade I-II aGVHD, and one developed grade IV. 2 patients occurred chronic GVHD (cGVHD), out of which one with local cGVHD and the other with extensive. 4 patients occurred graft rejection (GR), all of them recovered haemopoiesis and survived after donor PBSC infusion. 5 patients (12.2%) died, out of which one died of extensive cGVHD, and 4 died of invasive fungal infections (IFI). Median follow-up time was 23 (3 - 79) months. 36 patients survived. 5-year estimated overall survival (OS), disease free survival (DFS), and transplant-related mortality (TRM) was (81.1 ± 9.0)%, (68.4 ± 11.0)%, and (18.9 ± 9.0)%, respectively. Univariate analysis showed that lover OS had significant correlation with receiving PBSCT, occurrence of aGVHD, the number of infused CD34(+) cells no more than 2.5×10(6)/kg, the number of red blood cell (RBC) transfusion before transplant more than 30 U and occurrence of IFI after transplantation (P = 0.034, 0.001, 0.006, 0.000, 0.001, respectively). Occurrence of aGVHD had significant correlation with the disparity between donor and recipient ABO blood groups, the number of PLT transfusion more than 100 U, and the number of RBC transfusion more than 30 U before transplantation, the number of infused CD34(+) cells no more than 2.5× 10(6)/kg (P = 0.019, 0.038, 0.005, 0.005, respectively). The occurrence of GR had significant correlation with the number of PLT transfusion more than 100 U before transplantation (P = 0.038).

CONCLUSIONMSD allo-HSCT is an effective therapy for patients with SAA. Lower number of blood transfusion before transplantation, use of BMT, more number of infused CD34(+) cells can effectively prevent and treat aGVHD and IFI after transplantation, which may improve the efficacy of MSD allo-HSCT for SAA.

Adolescent ; Adult ; Anemia, Aplastic ; therapy ; Child ; Child, Preschool ; Female ; HLA Antigens ; Hematopoietic Stem Cell Transplantation ; Humans ; Male ; Retrospective Studies ; Siblings ; Tissue Donors ; Treatment Outcome ; Young Adult

BACKGROUNDWeaning difficulties occur in 31% of total intubated patients, and result in prolonged weaning duration. A computer-driven automated weaning system can perform a spontaneous breathing trial (SBT) automatically and display a message when the trial is successfully passed. Such a system might have a beneficial effect on difficult-to-wean patients. The aim of this study was to examine whether the computer-driven automated weaning system can accelerate discontinuation of mechanical ventilation and improve outcomes in difficult-to-wean patients.

METHODSThis randomized controlled study included 39 difficult-to-wean patients who failed their first spontaneous breathing trial. Before initiating weaning, eligible patients were randomly allocated to wean by computer-driven automated weaning system (CDW group, n = 19) or a physician-controlled local protocol (PW group, n = 20). Weaning duration, defined as the time from inclusion until first extubation, was the primary endpoint. Secondary endpoints were total duration of mechanical ventilation, the length of intensive care unit (ICU) stay, the number of reintubations, the mortality rate in the ICU, the number of noninvasive ventilations, the number of complications in the ICU, and the number of ventilator-associated pneumonia cases.

RESULTSThe weaning duration was reduced with the computer-driven weaning as compared with the usual protocol (median 29.0 hours vs. 45.5 hours, P = 0.044). Total duration of mechanical ventilation and duration of the ICU stay did not differ between the CDW and PW groups. There was no difference in the number of reintubations between the CDW and PW groups (3 and 4 patients, P = 0.732). The study groups showed comparable numbers of tracheostomy, self-extubations, ventilator-associated pneumonia, and non-invasive ventilation. Mortality in the ICU was similar in the CDW and the PW groups (21.1% vs. 20.0%, P = 0.935).

CONCLUSIONThe computer-driven automated weaning system can reduce weaning duration in difficult-to-wean patients as compared with a physician-controlled weaning protocol.

Aged ; Female ; Humans ; Intensive Care Units ; Male ; Middle Aged ; Respiration, Artificial ; methods ; Ventilator Weaning ; methods

OBJECTIVETo investigate the influencing factors of delayed gastric emptying (DGE) after pylorus-preserving pancreaticoduodenectomy (PPPD) and its preventing managements.

METHODSForty-two patients who underwent PPPD and 104 patients underwent standard Whipple procedure (SPD) between January 2000 and July 2006 were investigated retrospectively. The factors influencing the development of DGE following PPPD were analyzed statistically.

RESULTSThere was no significant difference in mortality between PPPD and SPD (0/42 vs. 7/104, P = 0.193). Pancreatic fistula occurred much more frequently in SPD than in PPPD (29/104 vs. 1/42, P < 0.05). The occurrence of DGE after PPPD was higher significantly than that after SPD (35.7% vs. 18.3%, P = 0.024). More DGE occurred in patients with a operation time more than 6 hours than in patients whose operation lasted less than 6 hours (76.9% vs. 17.2%, P = 0.008). Meanwhile, DGE occurred in 20% of patients with the antero-colonic route for duodenojejunostomy and in 50% with retro-colonic route (P = 0.043). Multivariate analysis by logistic regression model showed postoperative intra-abdominal complications were not risk factors for DGE. Prophylactic use of somatostatin couldn't prevent DGE effectively.

CONCLUSIONSDGE is the most frequent postoperative complication after PPPD, it can be markedly reduced by shortening operative time and using antero-colonic duodenojejunostomy procedure. There is no medicine which could prevent DGE effectively.

Aged ; Anastomosis, Surgical ; methods ; Duodenum ; surgery ; Female ; Gastric Emptying ; Humans ; Jejunum ; surgery ; Male ; Middle Aged ; Pancreaticoduodenectomy ; adverse effects ; methods ; Postoperative Complications ; etiology ; physiopathology ; Pylorus ; Retrospective Studies ; Risk Factors ; Stomach Diseases ; etiology ; physiopathology