The specific ScFv gene against botulinum neurotoxin A (BoNTa)was cloned into pPIC9k. Positive integrators were screened by increasing the dose of G418 in culture and expressed in Pichia pastoris GS115. As a result, engineered recombinant clone were obtained. 26 kD product of interest was seen easily in SDS-PAGE. Expression of human ScFv got the highest level 15% of total secreted proteins during 72～84 h after 1% methanol inducing. Purification of ScFv was finished by two steps: gel filter and ion exchange. Competing ELISA showed that recombinant ScFv could compete with antiserum to specific bind BoNTa.

Objective To explore the role of monitoring the cellular immune function in preventing and treating the fungal infection in the recipients of liver transplantation. Methods 679 cadaveric liver transplantations (from Jan. 2004 to Jan. 2010) were retrospectively studied. All the cases were divided into 3 groups according to different treatments and preventing regimens. The patients in groups A, B, C were treated based on the clinical experiences (394 cases), T lymphocyte subsets counting (151 cases), and combination of ATP values of CD4+ T cell and T lymphocyte subsets counting (134 cases), respectively. The infection, mortality and acute rejection rate were analyzed. The relationship between fungal infection and cellular immune function was investigated.Results The fungal infection rate in groups A, B, and C was 28. 9 %, 21.2 %, and 19. 4 % (P<0. 05), the morbidity rate was 16. 7 %, 12. 5%, and 3. 8% (P>0. 05), and the acute rejection rate was 28. 4 %, 17. 2 %, and 13. 4 % (P<0. 01), respectively. The CD4+ T lymphocyte counting in all cases of fungal infection was (147±43)×106/L. The morbidity could reach 50. 0 % when the CD4+ T lymphocyte counting < 100 ×106/L, while it was 2. 4 % when the counting was between (100-200) ×106/L (P<0. 01). The CD4+ T lymphocyte counting had no linear relation with the ATP value.The ATP value in fungal infection cases was (117 ± 61)μg/L. Conclusion The cellular immune function test could be quantitatively evaluated according to the T lymphocyte subsets and ATP value of CD4+ T lymphocyte. And individualized immunosuppressive therapy could be adjusted accordingly.Therefore, cellular immune function could be instructive in preventing and treating the fungal infection after liver transplantation.

ObjectiveTo investigate the features of the cortisol ( COR),thyroid hormone and autonomic function of anxiety disorder,and to observe the relationships between clinical subtypes,insomnia,gender factors and the neuroendocrine and autonomic function.Methods60 anxiety disorder patients and 60 healthy controls were randomly selected from a three-grade class A hospital over the same period.Blood at 8:00am to test the plasma COR,T3,T4,TSH level and examined the heart rate variability (HRV) on the same day,the anxiety disorder patients also finish the self-rating anxiety scale (SAS).ResultsThe plasma COR,TSH level and LF/HF index were significantly higher( (531.58 ± 218.59 ) nmol/L,P＜ 0.0l ; (3.50 ± 1.35 ) MIU/L,P＜ 0.05 ;3.87 ± 3.37,P＜0.0l ),while the SDNN,LF,HF index were significantly lower in anxiety disorder than that in the control group (27.34±13.13,P＜0.01;338.72 ±396.34,P＜0.01;135.77 ±149.95,P＜0.01).The plasma COR,TSH level and LF/HF index were significantly higher(P ＜ 0.05,P＜0.01,P＜0.01 ),while the SDNN and HF index were significantly lower in panic disorder than that in the generalized anxiety disorder (P ＜ 0.01 ).The plasma COR,TSH level and LF/HF index were significantly higher,while the SDNN,LF,HF index were significantly lower in panic disorder than that in the control group (P＜0.01 ).The plasma COR was significantly higher,while the SDNN and HF index were significantly lower in generalized anxiety disorder than that of the control group (P＜0.01 ).The plasma COR level and LF/HF index were significantly higher (P＜0.01,P＜0.05),while the SDNN,LF,HF index were significantly lower in anxiety disorder with or without insomnia than that in the control group (P＜ 0.01 ).The plasma TSH level was significantly higher in anxiety disorder with insomnia than that in the control group (P＜ 0.05 ).There were no significant difference of the neuroendocrine and autonomic function between anxiety disorder with or without insomnia (P ＞ 0.05 ).The plasma COR,TSH level were significantly higher,while the SDNN,LF,HF index were significantly lower in the female group than that in the male group (P＜0.01).The total SAS scores were significantly positively correhted with plasma COR levels,and was significantly negatively correlated with SDNN,HF index in the anxiety group.ConclusionThere are neuroendocrine and autonomic nervous system dysfunctions in anxiety disorder,and these dysfunctions are related with clinical subtypes,insomnia,and gender.

Objective To investigate the value of plasma H-FABP level and Acute Physiology and Chronic Health Evaluation(APACHE Ⅱ) in severity and prognosis estimation for patients with acute pumonary embolism(APE).Methods Totally 160 APE patients were hospitalized from January 2010 to January 2015 and enrolled in this study.According to the severity of the disease,these patients with APE were divided into low-risk group,moderate-risk group and high-risk group.According to clinical prognosis,these patients with APE were divided into survival groups and death groups.Plasma levels of H-FABP were measured by enzyme linked immunosorbent assay(ELISA),and APACHE Ⅱ score were analyzed.The differences of Plasma H-FABP levels and APACHE Ⅱ score were compared and which the relationship with severity and the prognosis of APE were also assessed.Results With the increased severity in patients,the H-FABP and APACHE Ⅱ score were significantly increased (P ＜ 0.05);the H-FABP and APACHE Ⅱ score were significantly higher in death group as compared with survival group(P ＜0.05).The H-FABP levels and APACHE Ⅱ score were positive correlated(r =0.71,P =0.000).ROC curves analysis results showed that the area under curve of H-FABP was 0.854 (95 ％ CI:0.784-0.927),and optimal operating point (OOP)was 13.3 μg/L,which had 81.0％ sensiticity and 79.4％ specificity;ACU of APACHE Ⅱ was 0.861 (95％ CI:0.812-0.932),and OOP was 19.2,which had 77.8％ sensiticity and 80.4％ specificity.The AUC was 0.914 (95％ CI:0.825-0.948),and the sensitivity was 88.9％,specificity was 87.6％ when the two cutoff values were both achieved,which were higher than the single H-FABP and APACHE Ⅱ score.Conclusion The H-FABP and APACHE Ⅱ score can effectively assess severity and prognosis of APE patients,meanwhile,it provide an objective basis for the clinical individual treatment and reducing the mortality rate of APE patients.

20ng/ml.The total three-year survival rate is 87%(33/35).Conclusion The effect and prognosis of ?-knife on prostate cancer treatment can be evaluated by the value of PSA before and after the treatment.

OBJECTIVETo analyze the related factors of the infection after operation of open tibiofibula fractures.

METHODSThe clinical data of 141 patients with open tibiofibula fractures underwent surgical treatment from June 2009 to December 2012 were retrospectively analyzed. All the patients were male and aged from 18 to 61 years old with an average of 39.2 years; all the fractures were unilateral. According to Gustilo typing of fracure, 5 cases were type I, 44 cases were type II, 27 cases were type III a, 56 cases were type III b and 9 cases were type III c. These clinical data included patients' age, gender, body mass index (BMI), underlying diseases, time of operation, fracture site, fracture type, fixation method, postoperative drainage, debridement condition and so on. The postoperative infection conditions were recorded, and the correlation between above factors and infections were analyzed. Finally, the significant related variables were introduced into a Logistic regression model to evaluated their risk.

RESULTSAmong the 141 patients, 22 cases developed with infection (15.6%). The significant relative factors with infection contained fixation method, debridement condition, underlying diseases and postoperative drainage (P < 0.05). The correlativities were stepped up in order, their odds ratios value was 2.451, 3.164, 3.414, 5.117, respectively.

CONCLUSIONActive treatment for underlying diseases before operation, thorough debridement for open wound, suitable fixation method for fracture and thorough draining are effective measures in preventing postoperative infection.

Adolescent ; Adult ; Debridement ; Fibula ; injuries ; Fractures, Open ; surgery ; Humans ; Logistic Models ; Male ; Middle Aged ; Postoperative Complications ; etiology ; Retrospective Studies ; Risk Factors ; Surgical Wound Infection ; etiology ; Tibial Fractures ; surgery

Tangcao pill is commonly applied in adjuvant and even alternative therapy for patients with AIDS. However, the herb contains complex ingredients, but with unknown effect against anti-HIV drug and unknown function. Because CYP450 emzyme is the main metabolic enzymes of the drug, it is of important significance to study the regulation of CYP450 enzymes before and after the combined administration of Tangcao pill and EFV. Proteomics, due to its high throughout and high sensitivity, has been widely applied in CYP450 enzyme study. In this paper, liver microsomes were separated through differential centrifugation. Their proteins were separated through SDS-PAGE. The three protein bands that CYP450 enzymes were located were cut and identified by liquid chromatography tandem mass spectrometry. Totally 16 CYP450 isoenzymes were identified. Furthermore, in order to make a quantitative analysis on the effect of tang herb on CYP450 emzyme, the multiple reaction monitoring (MRM) technology based on MS was adopted. The CYP2C11 was selected based on the results of the mass spectrum identification of proteins. The characteristic polypeptides were obtained through searching Expasy blast database. The m/z of the fragment ions was less than 800. In the paper, the m/z of ion pairs of CYP2C11 were 711.5/232.1, 711.5/319.2, 711.5/466.2 and 711.5/595.3, and the m/z of ESAT-6 (internal standard, IS) were 735.5/215.3, 735.5/389.3, 735.5/460.3 and 735.5/524.3. The relative peak (analyte/IS) area was adopted for the relative quantitative analysis. Compared with the EFV single administration group, the EFV and Tangcao pill combined administration group showed a 1.6-fold increase in CYP2C11. The results of the paper indicated that Tangcao pill may affect drug metabolism by regulating metabolic enzymes such as CYP2C11, but the specific mechanism still unknown.
Animals ; Cytochrome P-450 Enzyme System ; chemistry ; genetics ; metabolism ; Drugs, Chinese Herbal ; administration & dosage ; Electrophoresis, Polyacrylamide Gel ; Male ; Microsomes, Liver ; chemistry ; drug effects ; enzymology ; Proteomics ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo summarize the features and treatment of male infertility induced by autosomal dominant polycystic kidney disease (ADPKD), and compare the outcomes of intracytoplasmic sperm injection (ICSI) for infertile men with ADPKD and those with congenital bilateral absence of vas deferens (CBAVD).

METHODSWe retrospectively analyzed 21 cases of ADPKD-induced infertility, 15 treated by ICSI (group A), and another 164 cases of strictly matched CBAVD-induced infertility (group B). We compared the two groups in the couples' age, the number of ICSI oocytes, and the rates of fertilization, transferrable embryos, good embryos, embryos implanted, clinical pregnancy, biochemical pregnancy, early abortion, singleton and twins in the first cycle.

RESULTSAfter 28 cycles of ICSI, 10 of the 15 ADPKD-induced infertility patients achieved clinical pregnancy, including 7 cases of live birth, 1 case of spontaneous abortion, and 2 cases of pregnancy maintenance. No significant differences were observed between groups A and B in the couples' age, the wives' BMI, or the numbers of ICSI oocytes and embryos transplanted (P >0.05), nor in the rates of ICSI fertilization (72.64% vs 76.17%), transferrable embryos (51.28% vs 63.24%), quality embryos (38.46% vs 49.83%), embryo implantation (17.64% vs 38.50%), abortion (0 vs 9.23%), singleton (50% vs 81.54%) and twins (50% vs 18.46%). However, the rates of clinical pregnancy (13.33% vs 42.68%, P = 0.023 <0.05) and biochemical pregnancy (13.33% vs 39.63%, P = 0.032 <0.05) were significantly lower in group A than in B.

CONCLUSIONICSI is effective in the treatment of male infertility induced by either ADPKD or CBAVD, but the ADPKD cases have a lower success rate than the CBAVD cases in an individual cycle. The affected couples should be informed of the necessity of prenatal genetic diagnosis before embryo implantation and the inevitable vertical transmission of genetic problems to the offspring.

Abortion, Spontaneous ; Embryo Implantation ; Embryo Transfer ; Female ; Humans ; Infertility, Male ; therapy ; Male ; Male Urogenital Diseases ; therapy ; Oocytes ; Polycystic Kidney, Autosomal Dominant ; complications ; Pregnancy ; Retrospective Studies ; Sperm Injections, Intracytoplasmic ; Vas Deferens ; abnormalities

OBJECTIVETo study the combination of Mandibular distraction and orthognathic techniques for the reconstruction of adult hemifacial microsomia.

METHODSThe three-dimensional CT reconstruction data was used with Mimics for preoperation design. The osteotomy location, distraction vector, distraction distance were decided before operation with a surgical guider. At the first stage, internal distractor was implanted after ostetomy through an extra-oral approach. The distraction begun 5-7 days after operation with a frequency of 1 mm/day. After distraction, the distractor was maintained for 3-6 months. At the second stage, the distractor was removed. Le Fort I osteotomy was performed in order to correct the cross-bite and improve the facial contour. Usually, bone graft was inserted into the gap after Le Fort I osteotomy. The genioplasty was also performed if necessary.

RESULTS9 cases of adult hemifacial microsomia with severe mandibular deviation were treated. The facial asymmetry were improved greatly. 1 patient suffered an wound infection in the maxillary region after Le Fort I osteotomy and healed uneventfully with wound irrigation.

CONCLUSIONSMandibular distraction combined with orthognathic surgery is an effective procedure for adult hemifacial microsomia with complicated mandibular hypoplasia.

Adult ; Aged ; Bone Transplantation ; Facial Asymmetry ; surgery ; Goldenhar Syndrome ; surgery ; Humans ; Mandible ; surgery ; Osteogenesis, Distraction ; methods ; Osteotomy, Le Fort ; methods

The COI gene as DNA barcode was used to identify the Manis pentadactyla and its adulterants in order to provide a scientific basis for the molecular identification of M. pentadactyla. Genomic DNA was extracted from experimental samples using the DNA extraction kit. The COI genes were amplified using polymerase chain reaction (PCR) and sequenced bi-directionally. Obtained sequences were assembled using the CodonCode Aligner. The neighbor-joining (NJ) tree was constructed by MEGA 6.0. The results indicated that COI sequences were successfully amplified and NJ trees results indicated that M. pentadactyla and its adulterants can be easily identification. Therefore, the COI gene is an efficient barcode for identification of M. pentadactyla and its adulterants,which will provide a new technique for the market supervision.
Animals ; Cattle ; DNA Barcoding, Taxonomic ; methods ; Drug Contamination ; prevention & control ; Electron Transport Complex IV ; genetics ; Mammals ; classification ; genetics ; Medicine, Chinese Traditional ; Molecular Sequence Data ; Phylogeny ; Quality Control ; Sheep ; Swine