Factor VII Deficiency ; genetics ; Humans

To compare the activity of RGD-TRAIL in different expression systems, RGD-TRAIL in both Escherichia coli (E.coli) and Pichia pastoris was constructed and expressed. In vitro activity of RGD-TRAIL from Pichia pastoris expression system was also analyzed. Genetic engineering techniques were used to construct recombinant plasmid pET30-rgd-trail and pHBM-rgd-trail. The recombinant protein RGD-TRAIL was purified with Ni ion affinity chromatography after induction. MTT assay, ELISA, scratch wound healing, transwell migration assay and Hoechst 33342 staining were performed to detect the effects of RGD-TRAIL on proliferation, binding activity, migration and apoptosis. The expression of apoptosis-associated proteins was detected by Western blotting. Recombinant protein RGD-TRAIL was successfully expressed in a form of inclusion body in E.coli, while expressed secretorily in Pichia pastoris. It possessed more potent cytotoxicity than RGD-TRAIL in E.coli by MTT assay. The RGD-TRAIL expressed by Pichia pastoris showed powerful binding affinity with cancer cells expressing α(v), DR4, DR5 and highly potent cytotoxicity through inducing apoptosis of cancer cells. Nuclear fragmentation was examined by Hoechst 33342 staining. Cleaved PARP and caspase-3 were also detected after incubation with RGD-TRAIL. Additionally, RGD-TRAIL inhibited migration significantly in A549 and HT1080 cells. The results demonstrate that Pichia pastoris expression system is more suitable for the recombinant protein RGD-TRAIL. Its binding affinity and antitumor activity might make RGD-TRAIL a promising candidate for cancer therapy.
Antineoplastic Agents ; pharmacology ; Apoptosis ; Blotting, Western ; Cell Line, Tumor ; Chromatography, Affinity ; Enzyme-Linked Immunosorbent Assay ; Escherichia coli ; Humans ; Oligopeptides ; biosynthesis ; pharmacology ; Pichia ; metabolism ; Plasmids ; Recombinant Fusion Proteins ; biosynthesis ; pharmacology ; TNF-Related Apoptosis-Inducing Ligand ; biosynthesis ; pharmacology

Objective To evaluate the impact of epidural anesthesia with levobupivacaine combined with general anesthesia on colon surgery. Methods Sixty patients undergoing elective radical procedure for colon carcinoma were randomLy divided into four groups: saline group (group S), 0.125% levobupivacaine group (group L1), 0.25% levobupivacaine group (group L2), and 0.5% levobupivacaine group (group L3). Group S received normal saline of 10 mL epidurally and then infusion of 5 mL·h-1 until the procedure was finished; groups L1, L2, and L3 received levobupivacaine instead. Anesthetic induction was performed after epidural puncture. Mean blood pressure and heart rate were recorded at 8 time points including 5 min after entering into the operation room, 1 min after intubation, skin incision, abdominal exploration, 1 h after skin incision, completion of operation, extubation, and leaving PACU; meanwhile blood glucose and cortisol were detected, anesthesia time, time to PACU stay, bleeding, transfusion volume, adverse reaction, and doses of propofol, remifentanil, ephedrine, and fentanyl were noted. Results Time to PACU stay was longer in S group than in other 3 groups. Doses of remifentanil and fentanyl were larger in L1 group than in L2 group and L3 group. Ephedrine dose in L3 group was larger than in other 3 groups. Blood sugar in L1 group was higher than L2 group and L3 group. Cortisol in S group was higher than in other 3 group. Cortisol in L1 group was higher than in L3 group. The number of patients with hypotension was greater in L3 group than other 3 groups. Conclusions Continue epidural infusion of 0.25%levobupivacaine can reduce stress response and opioid uses, shorten PACU stay, whereas it does not increase use of ephedrine.

OBJECTIVESTo evaluate the effects of varicocele (VC) on IL-1 and NO levels in testes of rats with VC.

METHODSMale adult Wistar rats were divided into two groups randomly, VC group (n = 30) and pseudo-operation group (n = 20), and the levels of IL-1 and NO in the testes were determined and compared.

RESULTSThe levels of IL-1 and No of left tests in VC group were higher than those in pseudo-operation group, respectively(P < 0.01). While the levels of IL-1 and NO of right testes between two groups were not significantly different (P > 0.05). More over, the level of IL-1 correlated significantly with that of NO(r = 0.572, P < 0.01).

CONCLUSIONSThe results revealed that the changes of IL-1 and NO levels in the testes of rats with VC might be the reason which caused testes damage, disturbance of spermatogenesis and even infertility.

Animals ; Disease Models, Animal ; Interleukin-1 ; metabolism ; Male ; Nitric Oxide ; metabolism ; Rats ; Rats, Wistar ; Testis ; metabolism ; ultrastructure ; Varicocele ; metabolism

Cerebral Palsy ; classification ; etiology ; psychology ; Child, Preschool ; Humans ; Infant ; Intelligence ; Intelligence Tests ; Risk Factors

Objective To evaluate the efficacy of anesthesia with different doses of dexmedetomidine combined with propofol and remifentanil in patients undergoing abdominal surgery.Methods Ninety ASA Ⅰ or Ⅱ patients of both sexes,aged 18-64 yr,weighing 45-80 kg,undergoing abdominal surgery,were randomly assigned into 3 groups ( n =30 each):control group (group C ),dexmedetomidine 0.25 μg· kg-1 · h-1 group ( group D1 ) and dexmedetomidine 0.50 μg· kg-1·h-1 group (group D2 ).Dexmedetomidine was infused at a rate of 0.25 μg·kg-1 ·h-1 and 0.50 μg·kg-1 ·h-1 in groups D1 and D2 respectively until extubation after a loading dose of 0.5 μg/kg over 15 min.In group C,normal saline was infused intravenously at a rate of 10 ml/h.Anesthesia was induced with TCI of propofol with the target plasma concentration (Cp) of 1.0 μg/ml,iv injection of cisatracurium 0.2 mg/kg and TCI of remifentanil with Cp of 3 ng/ml.The patients were tracheal intubated and mechanically ventilated.PETCO2 was maintained 35-40 mm Hg and SpO2 was maintained ≥95％.Anesthesia was maintained with TCI of remifentanil with the target Cp of 5 ng/ml,iv infusion of cisatracurium 0.1 mg· kg-1 · h-1 and TCI of propofol.The target Cp of propofol was increased to maintain Narcotrend index of 37-46.The amount of remifentanil,cisatracurium and propofol consumed,extubation time and eye-opening time,complications during operation and during recovery from anesthesia were recorded.Results There was no significant difference in the amount of remifentanil and cisatracurium consumed and extubation time among the three groups ( P ＞ 0.05).Compared with group C,the eye-opening time was significantly prolonged,the incidence of hypertension and tachycardia during operation,and restlessness,vomitting,hypertension,and tachycardia during recovery from anesthesia was significantly decreased in groups D1 and D2,and the amount of propofol consumed was significantly decreased in group D2 (P ＜ 0.05).Compared with group D1,the eye-opening time was significantly prolonged,the incidence of hypertension during operation,and restlessness,hypertension,and tachycardia during recovery from anesthesia was significantly decreased in group D2 ( P ＜ 0.05).Conclusion When combined with propofol and remifentanil,dexmedetomidine infused at a rate of 0.50 μg·kg-1 · h-1 can provide satisfactory efficacy for abdominal surgery.

Objective To study the influence of pre-injection of donor apoptotic cells in the survival of islet grafts and the function of T lymphocytes in the peripheral blood. Methods The donor apoptotic cells and necrotic cells were ob-tained respectively by X-irradiation from electron linear accelerator and a heat-shock procedure (water bath box 56℃, 1 h). The diabetic rats for islet transplantation (n=42) were induced by a single intraperitoneal injection of streptozotocin (STZ), then were randomly divided into four groups:rats were injected by physiological saline group (n=9), normal cells group (n=12), apoptotic donor cell group (n=12) and necrotic donor cell group (n=9). On the seventh day, each group received islet transplantation under the renal capsule. The blood glucose level was detected to reflect the survival of the islets. The periph-eral blood samples of three rats in each group were obtained at different observation times. The proliferative activity of T lym-phocytes was determined by MTT method. The levels of cytokines interferon (IFN)-γ, interleukin (IL)-10 in peripheral blood were measured by Luminex 100 Integrated System, and transforming growth factor (TGF)-β1 by ELISA respectively at 0 d, 1 week, 2 weeks and after rejection. Results The survival time of islets was significantly prolonged by the pre-intervention of apoptotic cells, and the proliferative activity of T lymphocytes stimulated by ConA was inhibited. Meanwhile, the extent of the increased level of IFN-γwas inhibited significantly at 1 week and 2 weeks after transplantation (P<0.05), the levels of IL-10 and TGF-β1 were significantly increased before transplantation, 1 week and 2 weeks after transplantation (P<0.05). Conclusion Our results demonstrated that the pre-treatment of donor apoptotic cells can regulate the recipient’s immune reactive state by inhibiting the proliferative activity of T lymphocytes and changing the levels of cytokines from different sub-sets of T lymphocytes, and finally resulted in the prolonging of the survival of islet grafts.

Objective To identify the genotypes of ESBLs-producing Klebsiella pneumoniae isolates from the First Affiliated Hospital,Shantou University Medical College.Methods The MICs of 10 antibiotics were determined by agar-dilution against the clinical isolates of ESBLs-producing K.pneumoniae.PCR were performed with specific primers for blaTEM,blaSHV, blaCTX-M and blaOXA respectively.PCR products were cloned and sequenced.Results The results of PCR showed that a- mong the 83 strains of ESBLs-producing K.pneumoniae,75 were positive for blaTEM,41 positive for blaSHV,25 poitive for blaCTX-M,9 positive for hlaOXA.Three genotypes were found in 13 strains(15.7%),2 genotypes in 59 strains (71.1%) and single genotype in only 11 strains(13.2%).The genes of CTX-M-3,TEM-1 and SHV were found co-existent in 9 strains. The strains carrying 2 or 3 ESBL genes were more resistant to antibiotics than those carrying only 1 ESBL gene.Conclusions The genotypes of ESBLs-producing Klebsiella pneumoniae in this hospital are blaTEM,blaSHV,blaCTX-M and blaOXA. Most strains carry 2 or 3 ESBL genes.

To discuss the effective method of decreasing the postoperative recurrence rate of recurrent pterygium.
●METHODS:Totally 126 cases (126 eyes) with recurrent pterygium were randomly divided into A group (56 cases) and B group ( 70 cases ). Group A was treated by pterygium conjunctive reverse transplantation combined with amniotic membrane transplantation, group B was treated by amniotic membrane transplantation. The followed-up time after surgery was 6-24mo.
●RESULTS:ln group A, postoperative 5-7d (average 5. 62± 1. 38d), cornea epithelium was repaired. ln group B, postoperative 7- 10d ( average 7. 38 ± 1. 12d), the corneal wound was healed. There was statistical significant difference between two groups (t = 4. 307,P<0. 05). Three cases recurrence were noted in A therapeutic group (56 cases), the recurrent rate was 5. 4%; Twelve cases recurrence were noted in B compared group (70 cases), the recurrent rate was 17. 1%. There was statistical significant difference between two groups(P<0. 05).
●CONCLUSlON: lt is suggested that pterygium conjunctive reverse transplantation combined with amniotic membrane transplantation is effective in the treatment of recurrent pterygium.