Objective 4-Hydroxyisoleucine (4-HIL) is the active component isolated from the seeds of Trigonella foenum-graecum. It exerts broad pharmacological effects such as anti-hyperglycemia,blood lipid lowing,improvement of insulin resistance,etc. As the Chinese traditional medicine,it can be used clinically in treating many diseases such as type 2 diabetes mellitus,dyslipidemia,metabolic syndrome,fatty liver and other diseases. Here the current research status and application prospect on 4-Hydroxyisoleucine were summarized.

Objective To investigate the epidemiological features of out-of-hospital cardiac arrest (OHCA) in Zhejiang and to analysis factors associated with outcomes for providing evidence on improving the success rate of cardiopulmonary resuscitation (CPR). Methods Clinical data of 493 patients with OHCA collected from the emergency department (ED) of Zhejiang Provincial People's Hospital, Ningbo Emergency Medical Service Center and Shaoxing Emergency Medical Service Center from January 2012 to August 2016 were analyzed retrospectively. All the data were recorded following the Utstein style included causes of arrest, location of arrest, first arrest rhythm, witnessed by bystanders, bystander CPR, pre-hospital defibrillation, pre-hospital intubation, pre-hospital epinephrine administration, emergency medical services (EMS) response time, return of spontaneous circulation (ROSC) at scene/enroute, ROSC at ED, admission to hospital, survival to hospital discharge and neurological outcomes at discharge. Factors associated with 30 days survival rate of patients with OHCA were analyzed. Results 493 patients were enrolled, of whom 342 were male and 151 were female. The average age was (58.8±21.4) years. The causes of arrests consisted of cardiac etiology (219 cases), trauma (155 cases), respiratory disease (22 cases), drowning (19 cases), electrocution (8 cases) and others (70 cases). Most of the events occurred at home (65.1%) and public places (22.7%). 55.2% patients were witnessed by bystanders while bystander CPR was performed in only 2.6% cases. Asystole was the predominant rhythm (78.7%) observed by the ambulance crew at the arrest site while only 5.5% first arrest rhythms were ventricular fibrillation/ventricular tachycardia (VF/VT). Only 6.9% patients underwent pre-hospital defibrillation. Pre-hospital intubations were attempted in 16.4% patients. Epinephrine was administered in 56.4% patients at scene or in ambulances. EMS response time was (13.6±8.0) minutes. 4.5% patients had ROSC at scene/enroute and 7.7% had ROSC at ED. Only 9.7% patients were admitted to hospital and 1.2% discharged alive. 0.8% patients were still in hospital on 30th day. The 30-day survival rate was 2.0% (10/493) and only 0.8% patients had neurologically favorable survivals [with the cerebral performance category (CPC) score of 1 or 2]. Witnessed by bystanders (3.31% vs. 0.45%), VF/VT as the first arrest rhythm (7.41% vs. 1.72%), bystander CPR (15.38% vs. 1.67%), pre-hospital defibrillation (8.82% vs. 1.53%) and EMS response time < 10 minutes (3.57% vs. 0.74%) could improve 30-day survival rate of OHCA significantly (all P < 0.05). Conclusion Resuscitation survival of OHCA in Zhejiang province was unsatisfactory. Improvements are required in series aspects of OHCA survival chain.

Objective To observe the effect of gastrectomy for gastric cancer on blood glucose in patients complicated with type 2 diabetes mellitus by using different operation strategies.Methods The retrospective analysis was carried out of 68 cases who undergone radical surgery for gastric cancer and complicated with type 2 diabetes mellitus.These patients were divided into 3 groups by different kinds of digestive tract reconstruction,Billroth Ⅰ (n =26),Billroth Ⅱ (n =22) and Roux-en-Y (n =20).The levels of fasting blood glucose (FBG) were detected before and after operation one week,one month and six months in three groups.Results Comparing to the level before operation,the level of FBG after operation showed a descent in group B and group C((10.03 ±5.12) vs.(13.56 ±3.36) mmol/L; (8.44 ±5.21) vs.(13.56 ±3.36) mmol/L;(10.61 ±3.16) vs.(12.84 ±3.36) and(7.68 ±4.88) vs.(12.84 ±3.36) mmol/L) which was better than group A((10.03 ±5.12)vs.(11.12 ±6.32),(8.44 ±5.21) vs.(11.12 ±6.32) mmol/L;(10.61 ±3.16) vs.(11.12 ±6.32) mmol/L),(7.68 ±4.88) vs.(11.12 ±6.32) mmol/L;P＜0.05,but there was no significantly difference between B and group C (P ＞ 0.05).The 3 groups of patients,with B,C groups of patients with diabetes improved obviously,of which nearly 50％ of patients cured,normal blood sugar control,the other part with the exception of a few effective,degradation of diabetes drug treatment,and operation effect was significant.Conclusion It is effective to control the blood glucose of gastric cancer patients complicated with type 2 diabetes with Billroth II or Roux-en-Y gastrectomy.There is a correlation between postoperative blood glucose change and different gastrojejunal anastomosis.

Objective To evaluate the efficacy and safety of HLA identical and haploidentical related allogeneic hematopoietic stem cell trans?plantation in the treatment of acute myeloid leukemia(AML)and myelodysplastic syndrome(MDS). Methods A total of 21 patients with AML and 8 patients with MDS who underwent allogeneic hematopoietic stem cell transplantation in our hospital from April 2011 to April 2016 were ana?lyzed retrospectively,including 16 cases of HLA?identical allogeneic HSCT,10 cases of haploidentical allogeneic HSCT,and 3 cases of syngeneic HSCT. BUCY2 or TBI plus CY ± chemotherapeutic agents was the regular conditioning regimen. No graft versus host disease(GVHD)prophylax?is was required for syngeneic HSCT,but cyclosporine in combination with methotraxate was essential for allogeneic HSCT,cyclosporine,methotrax?ate,antithymocyte globulin,mycophenolate mofetil and glucosteroids for haploidentical HSCT. Results All patients achieved fully donor?originat?ed hematopoiesis. Two patients died of severe acute GVHD within 100 days post HSCT. Acute GVHD with gradeⅡ?Ⅳoccurred in 23.1%(6/26) patients,chronic GVHD in 50%patients,therapy and relapse?relevant mortality was 4/29(13.8%)and 6/29(20.7%)cases within a median follow?up of 23(1?60)months,respectively. Two?year overall survival and leukemia free survival rates are 68.09%( 95%CI:45.77%?82.78%)and 60.22%(95%CI:38.19%?76.55%),respectively. High risk AML is still the main challenge to long?term leukemia free survival. Conclusion HLA identical and haploidentical allogeneic HSCT for AML and MDS is safe ,effective and feasible. Minimal residual disease monitoring and pre?ventative as well as preemptive intervention is necessary for improving prognosis of high risk AML.

Objective To analyze the clinical manifestation of diffusive alveolar hemorrhage in acute leukemia induction therapy.Methods Clinical data of two diagnosed cases of diffusive alveolar hemorrhage secondary to acute leukemia were collected.Clinical data of eight cases of diffusive alveolar hemorrhage secondary to acute leukemia which were published were also collected by searching in Medline database.The clinical manifestation,diagnosis,strategy of differential diagnosis and treatment of diffusive alveolar hemorrhage secondary to acute leukemia were analyzed.Results Diffusive alveolar hemorrhage was a rare but fatal complication of acute leukemia.The common clinical manifestations included hemoptysis,progressive dyspnea and progressive decrease in concentration of hemoglobin.The analysis of blood gas showed type Ⅰ respiratory failure.The manifestations of chest computed tomography included diffusive ground glass opacity and infiltration of parenchyma.The bronchoalveolar lavage fluid was bloody.And lung biopsy showed congestion of alveoli and capillaritis.The detection for pathogens,vasculitis related antibodies,brain natrium peptide were negative.The mortality of those cases was 40 ％ (4/10).Corticosteroids therapy was effective.The mortality of patients received corticosteroids therapy was 25 ％ (2/8).Conclusion Diffusive alveolar hemorrhage is a rare but fatal complication of acute leukemia.The mortality is high.The key points of therapy are early diagnosis and corticosteroids therapy.

Objective To develop a novel anti-Enterovirus 71 (EV71) vaccine as recombinant virus-like particles.Methods By utilizing the foreign antigen presentation and virus-like particles forming features of Norovirus casipid VP1 P domain (NoVP), two pET-28a (+)-based recombinant expression plasmids containing either NoVP alone or NoVP with three specific epitopes SP55, SP70 and VP2-28 of EV71 capsid proteins tandemly inserted at the surface loop site were constructed and transferred to Escherichia coli.The recombinant fusion proteins of NoVP + EV71-SP55-SP70-VP2-28 and NoVP were induced expression and confirmed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), Western blot and transmission electron microscopy (TEM) observation.BALB/C mice were randomly divided into three groups:group A immunized with the recombinant fusion protein, group B immunized with NoVP and group C injected with 10 mmol/L Tris plus 20 mmol/L NaCl (pH 9.0).Enzymelinked immunosorbent assay (ELISA) was used to test specific antibodies in the serum of the mice, besides, the serums were mixed with the EV71 H3-TY strain and Vero cells, then specific antibody titer was examined by microneutralization test.One way ANOVA and Bonferroni test were used to analyze data.Results Both recombinant fusion protein and NoVP were expressed in Escherichia coli in inclusion bodies form.SDS-PAGE demonstrated that the relative molecular weights of recombinant fusion protein and NoVP protein were approximately 43 × 103 and 36 × 103, respectively;positive protein band of about 43 × 103 (relative molecular mass) was detected in recombinant fusion protein by Western Blot.Virus-like particles derived from the recombinant fusion proteins were observed under TEM.ELISA showed that absorbance 490 (A490) of mice serum added in SP70 peptide was significantly higher than those of group B and C (F =13.860,P ＜0.05).And microneutralization test demonstrated that the serum from group A was able to neutralize EV71 at a geometric mean titer above 1:38.Conclusion A novel virus-like particles vaccine against EV71 with good antigenicity and specificity has been prepared, which is able to induce high titer of neutralizing antibody against EV71 in mice.

BACKGROUND: In recent years, nano-carriers have been regarded as the most promising technologies for breakthrough the bottleneck of gene transfer. Polyamidoamine dendrimer (PAMAM) is a kind of new nanometer material. PAMAM can transfer target gene to the cell with high efficiency and lower toxic both in vivo and in vitro.OBJECTIVE: To evaluate the antitumour effects of survivin antisense oligonucleotide (Survivin-asODN) carried by PAMAM on colorectal cancer transplanted subcutaneously in nude mice.DESIGN, TIME AND SETTING: An in vivo experiment regarding tumor gene therapy was performed from February to August in 2008 at the Laboratory of Bionanometer Engineering, Research Institute of Micro/nanometer Science & Technology of Shanghai Jiao Tong University and Central Laboratory of Zhujiang Hospital of Southern Medical University.MATERIALS: Human colorectal cancer cells SW620 were from Shanghai Cell Institute of Chinese Academy of Sciences.PAMAM dendrimer was offered by the Bionanometer Engineering Laboratory, Research Institute of Micro/nanometer Science & Technology, Shanghai Jiao Tong University. Lipofectamine ~(TM)2000 was purchased from Invitrogen, USA. Survivin-asODN was synthesized by Shanghai Bioengineering Company.METHODS: The PAMAM and cation liposome were respectively mixed with Survivin-asODN to generate the transfection complex carrying antisense gene. The shape of the complex was observed by transmission electron microscope, the particle size was determined by laser particle size analysator and the zeta potential was measured by an analytical tool. The encapsulating efficiency and release progress in vitro were determined by ultraviolet spectrophotometer in centrifuging method. Human colorectal cancer cells SW620 at logarithmic phase were inoculated into the abdominal region of 18 Blab/C nude mice subcutaneously to produce transplanted tumor models in colorectal cancer nude mice, which were randomly divided into 3 groups: liposome, PAMAM and blank control groups. They were injected respectively with Hposome-survivin-asODN complex,PAMAM-survivin-as ODN transfection complex and seroculture liquid. The volumes of tumor were surveyed in the 2 groups.Western blotting method was used to determine the survivin gene expression in the transplanted tumor tissue.MAIN OUTCOME MEASURES: Particle size, zeta potential, gene loading level, encapsulation efficiency, release rate of cationic liposome-survivin-asODN complex and PAMAM-survivin-asODN complex, as well as survivin expression rate and apoptosis rate after transfection, inhibition rate of the transplanted tumor growth, Survivin protein expression and activity in the transplanted tumor cells.RESULTS: The particle size of PAMAM-survivin-asODN complex was smaller (P < 0.01), but the zeta potential was greater (P < 0.05), compared with liposome-survivin-asODN. There was no significant difference between PAMAM and liposome groups in terms of gene loading rate and transfection efficiency. DNA release lasted for 14 days for PAMAM, but only 5 days for liposome.After colorectal cancer cell transfection, survivin protein expression was lower, but apoptosis rate was higher, in the PAMAM-survivin-asODN complex than in the liposome-survivin-asODN complex (P < 0.05).CONCLUSION: PAMAM facilitates delivery of Survivin-asODN into transplanted colorectal cancer cells SW620. As a result,survivin protein expression was decreased, and apoptosis rate was increased in vivo which inhibited transplanied tumour growth.

AIM: To investigate the expression of vitamin D3 up-regulated protein 1 (VDUP1) in peripheral eosinophils of asthma patients and its relation with eosinophil activation.METHODS: 10 normal volunteers and 31 mild to moderate asthma patients were selected. Symptom severity, pulmonary function index, induced sputum eosinophil counts were recorded. Then, gene and protein expressions of VDUP1 and ?-actin were evaluated by RT-PCR and Western blotting, respectively. In addition, eosinophils were incubated with IL-5, both VDUP1 and ?-actin were amplified by RT-PCR. The eosinophil cationic protein (ECP) of supernatant and serum were also detected by ELISA assay. RESULTS: There was a significant decrease in expression of VDUP1 in asthma attack patients without treatment compared with normal volunteers and patients in remission. In contrast, no significant difference between the patients in remission and normal volunteers was observed. In patients with asthma attacks, a negative relationship between expression intensity of VDUP1 and EOS% in induced sputum and serum ECP concentration was also observed. The expression of VDUP1 in eosinophils was decreased by IL-5 stimulation, simultaneously, the ECP in supernatant was increased. CONCLUSION: The expression of VDUP1 in eosinophils decreases in asthma patients, and is negatively associated with serum ECP and induced sputum EOS%. EOS activation by IL-5 may be related to VDUP1 pathway.

BACKGROUND: Mesenchymal stem cells are the stem cells that possess the capability for self-renewal and multi-directional differentiation. Umbilical cord is the tissue outside the embryos and would be fallen off after parturition. In addition, it has wide source and no ethical restriction, so it is promising to be the first choice for mesenchymal stem cells. OBJECTIVE: To detect the surface markers CD29, CD44, CD49e, CD73, CD90, CD34, CD45, and CD271 of human umbilical cord-derived mesenchymal stem cells (hUCMSCs) prior to and after cryopreservation and resuscitation. METHODS: After isolation and culture, morphology of the primary, P4 and P8 hUCMSCs was observed prior to cryopreservation and after resuscitation. Surface markers CD29, CD44, CD49e, CD73, CD90, CD34, CD45, and CD271 of primary, P4, and P8 hUCMSCs were detected through the use of flow cytometry prior to cryopreservation and after resuscitation RESULTS AND CONCLUSION: hUCMSCs prior to cryopreservation and hUCMSCs of different passages after resuscitation present the same phenotype, i.e., positive for CD29, CD44, CD49e, CD73, and CD90, and negative for CD34, CD45, CD271. These findings suggest that primary hUCMSCs do not present changes in surface markers after cryopreservation and resuscitation.

The traditional fermentation engineering experiment requires a reform on the experimental con-tents and teaching pattern. According to the production process of industrial enzymes, we set up a two-week comprehensive experiment. The students designed and prepared the experiment by themselves. Moreover, the pattern of self-management was used in the process and the experiment scores included the self-assess-ment and objective assessment. It was proved that the new teaching pattern increased the study interesting of students, inspired their initiative and trained their spirits of team cooperation. The teaching effect was im-proved markedly and good ideas are also put forward to solve the possible problem.