Anesthetics, Intravenous ; administration & dosage ; pharmacology ; Animals ; Animals, Newborn ; Cerebral Cortex ; cytology ; metabolism ; Drug Synergism ; Female ; Fentanyl ; administration & dosage ; pharmacology ; Male ; Midazolam ; administration & dosage ; pharmacology ; Neurons ; metabolism ; Patch-Clamp Techniques ; Primary Cell Culture ; Rats ; Rats, Sprague-Dawley ; Voltage-Gated Sodium Channels ; drug effects

Methyl parathion hydrolase (MPH, E.C.3.1.8.1) coding gene mph from Pseudomonas sp. WBC-3, isolated and identified by our lab, was successfully expressed in E. coli AD494 (DE3)/ pET32a(+) system as soluble fusion form at high level. The recombinant MPH showed nearly 4～5 fold higher specific activity to parathion than the enzyme from Pseudomonas sp. WBC-3. In addition, the thermal stability of the recombinant enzyme was improved comparing with the wild type enzyme.

Objective: To explore the intervention effect of Yishenbaozhen Recipe (YSBZR) on the renal anemia in rats with chronic renal failure and the related mechanism. Methods: Totally 43 rat models of chronic renal failure were produced by two-step 5/6 nephrectomy. The study was randomly divided into 5 groups: normal control group (n=9), model group (n=8), YSBZR high-dose group (n=9, 46. 5 g/[kg • d]), YSBZR low-dose group (n=9, 15. 6 g/[kg • d]) and Niaoduqing group (n=8, 2.5 g/[kg • d]). YSBZR was given intragastrically on a daily bases for 6 weeks, and then the serum erythropoietin (EPO), serum creatinine (SCr), blood urea nitrogen (BUN), hemoglobin (Hb), red blood cell count (RBC), and mean corpuscular volume (MCV) were examined. Results: Compared with the model group and Niaoduqing group, YSBZR high-dose group and low-dose group had significantly higher Hb, RBC and EPO levels (P<0.05), and had similar MCV levels. YSBZR high-dose and low-dose groups had significantly decreased SCr and BUN levels compared with the modll group (P<0.01), and had similar levels when compared with Niaoduqing group. Conclusion: YSBZR has interventional effect on renal anemia, which might be related to decreasing nitrogen retention, reducing the toxin-induced inhibition of hematopoietic system, and increasing EPO level.

Objective To analyze the bacterial diversity in stem rot and healthy plants of Paris polyphylla var. chinensis. Methods Bacterial strains were isolated from rhizomes, stems and leaves of the diseased and healthy plants of P. polyphylla var. chinensis using beef extract-peptone medium. Using 16 S rRNA universal primers 27F/1492R for PCR amplification, combined with DNA sequencing technology to preliminary identification the bacterial strains. Results The bacteria could be divided into 23 microbial species, belonging to 11 genera, isolated from healthy and diseased plants of P. polyphylla var. chinensis. Eleven endophytic bacteria, belonging to 4 genera were contained from healthy plants, in which there were 9, 10, and 5 species isolated from rhizomes, stems and leaves, respectively. Fourteen endophytic bacteria, belonging to 10 genera were contained from diseased plant, in which there were 11, 8, and 3 species isolated from rhizomes, stems, and leaves, respectively. The content of endophytic bacteria in rhizome of healthy plant was the highest, reached up to 2.999 × 105 cfu/g, while that in leaf was the lowest with 7.32 × 104 cfu/g. The quantities of Bacillus species in rhizome (73.3%), stem (67.1%), and leaf (81.8%) of healthy plant were the highest groups, Pseudomonas species in rhizome (35.6%), stem (50.3%) and leaf (60.5%) of diseased plants were the highest groups. Shannon-Wiener index and evenness index of healthy plant of P. polyphylla var. chinensis were higher than that of diseased plants. Conclusion The dominant group in the healthy plant of P. polyphylla var. chinensis was Bacillus, while the dominant groups in the diseased plant of P. polyphylla var. chinensis was Pseudomonas. The population diversity of the cultivable bacteria in healthy plant of P. polyphylla var. chinensis was more abundant than that of diseased plant.

Objective: In this work, phylogenetic analysis was used to compare the ITS2 and psbA-trnH sequences of Polygonatum cyrtonema samples from different geographical sources, so as to explore the genetic diversity and genetic relationship of these resources. Methods: PCR method was used to amplify the regions of ITS and psbA-trnH, and the sequences of ITS2 and psbA-trnH were obtained after the amplified fragment sequences were blasted in NCBI database. The neighbor joining (NJ) and maximum parsimony (MP) methods were used to construct phylogenetic trees and Kimura two-parameter (K2-P) model was used to calculate the genetic distance of different samples. Mega and DNAman softwares were applied for mutiple alignment of ITS2 and psbA-trnH sequences of 25 samples of P. cyrtonema. Results: The lengths of ITS2 and psbA-trnH sequences of Anhui Qingyang and Fujian Taining samples of P. cyrtonema were 224 bp and 620 bp, respectively. The lengths of ITS2 and psbA-trnH of the remaining 24 samples were 225 bp and 621 bp, respectively. ITS2 and psbA-trnH had seven and four mutation points, respectively. These 25 samples were clustered into two groups based on ITS2 sequences. Five samples in Hunan and Guizhou were clustered into one group, while the other 20 samples were clustered into another group. The genetic distance showed that the samples from Huaxi and Jianhe in Guizhou Province and Jianyang in Fujian Province had the largest genetic distance. Phylogenetic tree constructed by psbA-trnH sequences were unable to distinguish 25 samples from different geographical sources. Conclusion: Phylogenetic and mutation analysis will provide the theoretic foundation to utilize the resources of P. cyrtonema, investigate their evolution, and evaluate their genuineness. The results of mutation point will also be used in the identification of related P. cyrtonema resources.

Objective To compare and analyze the effect of prophylactic postoperative radiotherapy for esophageal carcinoma.Methods 102 such patients were treated with prophylactic radiotherapy after radical resection,to a total dose of 50-60 Gy.The extensive portal included supraclavicular region on both sides,entire mediastinum,the site of anastomosis and left gastric lymph node region in 43 patients.The re- gional portal range was different according to the different location of primary lesion in 59 patients.Results The 1-,3- and 5-year survival rate was 76%,51% and 43% respectively,with a median survival of 30 months.The 1-,3- and 5-year survival rate was 77%,52% and 41% in the extensive portal and 76%, 49% and 45% in the regional portal,respectively(P=0.884).According to multivariate analysis,N stage, number of metastatic lymph nodes and tumor length were independent prognostic factors.Conclusions Regional portal does not lower the survival rate when prophylactic postoperative radiotherapy is used in e- sophageal carcinoma.

Objective:To investigate the effect of dexmedetomidine on stress response and cellular immune function in patients undergoing radical resection of colon cancer during perioperative period.Methods: 96 cases of colon cancer undergoing radical resection were randomly divided into dexmedetomidine group(group D)and control group(group C).The group D was intubated with in-travenous infusion of dexmedetomidine 0.6 μg/kg,followed by intravenous infusion of 0.3 μg/(kg·h) at the end of the surgery,and the group C was given 0.9% saline at equal volume and rate.The venous blood of patients was collected before anesthesia induction,10 min(T0),immediate postoperative(T1),postoperative 24 h(T2)and postoperative 72 h(T3).Flow cytometry were used to detect T lym-phocyte subsets(CD3+,CD4+,CD8+,CD4+/CD8+)and the percentage of NK cells,and determination of norepinephrine(NE), epinephrine(E),cortisol(Cor),IL-6,IL-10 and TNF-α levels.Records of patients with T0,immediate intubation(Ta),T1,immediate extubation(Tb)of SBP,DBP,MAP,and postoperative adverse reactions were recorded.Results:The SBP,DBP and MAP of group C at Ta,T1,Tbwere significantly higher than that of T0and group D(P<0.05).Compared to those at T0,the levels of CD3+,CD4+,CD8+and CD4+/CD8 at T1and T2were significantly lower in both groups(P<0.05),and the levels of group C at T1,T2,T3were significantly lower than those in group D(P<0.05).The NK cells of group C at T1,T2were significantly lower than that of T0and group D(P<0.05).Compared to those at T0,the levels of IL-6,IL-10 and TNF-α at T1and T2were significantly higher in both groups(P<0.05), and the levels in group C were significantly higher than those in group D(P<0.05).The IL-6 and IL-10 of group C at T3were significantly higher than that of T0and group D(P<0.05).Compared to those at T0,the levels of NE,E and Cor at T1and T2were sig-nificantly higher in both groups(P<0.05),and the levels in group C were significantly higher than those in group D(P<0.05).The NE,E and Cor of group C at T3were significantly higher than that of T0and group D(P<0.05).The proportion of adverse reactions in group D was significantly lower than that in group C(χ2= 4.800,P= 0.028).Conclusion: Dexmedetomidine can inhibit the perioperative stress response and reduce the impact on immune function in patients undergoing radical resection of colon cancer.

Objective To detect the mutations of ATP2C1 gene in patients with Hailey-Hailey dis- ease (HHD).Methods PCR and direct sequencing were performed in 17 patients and 120 healthy controls to screen the mutations in the exons of ATP2C1 gene.Results Eight mutations were identified in nine probands, including three deletion mutations (nt1464-1487 del/nt1462-1485del,1523delAT,2375delTTGT),three splice site mutations (360—2A→G,1415—2A→T,2243+2T→C) and two missence mutations (C920T and G1942T).None of the above mutations was found in the controls.Conclusion Eight specific novel mutations were identified in nine probands of HHD,which could be causative factors of the disease.

Objective To investigate the therapeutic method of metabolic acidosis in long-term sur- vival patients undergoing simultaneous pancreas and kidney transplantation.Methods A 45-year-old fe- male patient,who had undergone simultaneous pancreas and kidney transplantation(due to diabetic ne- phropathy and uremia)with bladder drainage 2 years before,developed severe metabolic acidosis,and thus underwent surgical conversion from bladder to ileal drainage.The procedure was as follows.The stoma of duo- denocystostomy was isolated and resected.The site of cystostomy was closed in two layers.The graft duode- num was then anastomosed to a loop of the recipient's ileum,which was proximal 40 cm from the ileocecum in a side-to-side manner.Results The metabolic acidosis resolved postoperatively.The patient received conventional immunosuppressants.The hospital stay was 30d.Follow-up of 4 years showed normal pancreas and kidney functions.Conclusions Conversion from bladder to ileal drainage is safe and effective for metabolic acidosis related to the exocrine secretions of bladder drained pancreas graft in simultaneous pancre- as and kidney transplant recipients.

Objective To define the maximum-tolerated dnse(MTD)and observe the side effect of escalating cisplatin with 5-fluorouracil in concurrent chemoradiotherapy for esophageal carcinoma in Chinese,with toxicity studied.Methods Previously untreated fifteen Chinese patients suffering from esophageal carcinoma received conventional fractionafiun radiotherapy,with 5 daily fractions of 2.0 Gy per week.The total radiation dose was 60 Gy.Concurrent chemotherapy dose escalation was given by the relatively safe and kidney-sparing modified Fibonacci sequence.The starting dose was cisplatin 37.5 mg/m~2 D1 and 5-fluorouracil 500 mg/m~2 D1-5, respectively.This regimen was repeated 4 times every 28 days.Escalation dose was eisplatin 7.5mg/m~2 and 5- fluorouracil 100mg/m~2.Every cohort contained at least 3 patients.If no dose-limiting toxicity(DLT)was observed, the next dose level was opened for entry.These courses were repeated until DLT appeared.MTD was declared as one dose level below which DLT appeared.Results DLT was defined as grade 3 radiation-induced esophngitis at the level of cisplatin 60 mg/m~2,5-fluorouracil 700 mg/m~2.MTD was defined as eisplafin 52.5 mg,/m~,5- fluorouracil 700 mg/m~2.The major side effect were radiation-induced esophagitis,leucopenia,nausea,vomiting and anorexia.Conclusion Maximun tolerated dose of cisplatin with 5-fluorouracil in concurrent chemoradiotherapy in the Chinese people with esophageal carcinoma were eisplatin 52.5 mg/m~2 D1,5-fluorouracil 700 mg/m~2 D1-5,repeated 4 times every 28 days.