Animals ; Birds ; Disease Outbreaks ; prevention & control ; Hong Kong ; epidemiology ; Humans ; Influenza in Birds ; epidemiology ; prevention & control ; Poultry ; Public Health ; Public Health Administration

Objective To observe the effect of community rehabilitation on the motor function and activities of daily living(ADL)of stroke patients.Methods 155 stroke patients were randomly divided into the exercises group(n=76)and control group(n=79)and evaluated with simple Fugl-Meyer Assessment(FMA),Modified Barthel Index(MBI)and Functional Comprehensive Assessment(FCA)before treatment and 1 and 3 months after treatment.Results The evaluations of FMA,MBI and FCA were not different between two groups before treatment.One month later,the scores of MBI and FCA of the exercises group improved significantly(P<0.01),but simple FMA not improved even to 3 months after treatment(P>0.05).In the control group,the simple FMA score not improved(P>0.05),the scores of MBI and FCA had a declined tendency with disease course prolong.Conclusion The effect of community rehabilitation on the motor function of stroke patients during convalescence is not significant,but community rehabilitation may improve ADL and the quality of life.

Objective To construct the prokaryotic expression vector of mouse CD25 extracellular domain and to express it in E coli.Methods Total RNA was isolated from splenocytes of Balb/c mice.The CD25 extracellular domain gene was amplified by RT-PCR and cloned into the PET-32a vector.A positive recombinant,PET-32a-CD25e,was identified by enzyme cleaving and sequencing before its expression in E.coli,and transferred into E.coli BL21(DE3) plysS for its expression.After purification with Ni+ resin and renaturation in vitro,a relative molecular mass(Mr) of the interesting protein was detected by SDS-PAGE and Wes-tern blotting.Effect of the purified interesting protein on the proliferation of splenocytes from T cell vaccine-immunized syngeneic mice was detected by MTT assay.Results The cloned CD25 extracellular domain gene was identified to be functional by sequencing and expression.The purified interesting protein could significantly induce the proliferation and IL-4 secretion of splenocytes from T cell vaccine-immunized mice in vitro.Conclusion Mouse CD25 extracellular domain gene can be successfully expressed in prokaryotic cells with biological activity,which lays a foundation for further relative studies.

Acute Disease ; Adult ; Benzene ; poisoning ; Female ; Humans ; Leukemia ; chemically induced ; Occupational Exposure ; adverse effects

Postoperative pancreatic leakage remains a persistent problem after pancreaticoduodenectomy.For patients with a soft and nonfibrotic pancreas,double binding continuous hemstitch suture is an optimal method for anastomosis.From January 2011 to June 2012,92 cases of periampullary carcinoma with a soft pancreas underwent pancreaticoduodenectomy,and then a modified technique of pancreaticogastrostomy was performed with 2 continuous hemstitch sutures placed in the mucosal and seromuscular layers of the posterior gastric wall,respectively.The median time for pancreaticogastrostomy was 12 minutes,and only 15 patients had postoperative complications.Two patients developed pancreatic leakage(1 grade A and 1 grade B)postoperatively.Pancreaticogastrostomy with double binding continuous hemstitch sutures is a simple and safe reconstruction procedure for patients with a soft and fragile pancreas who received pancreaticoduodenectomy.

Objective To construct the genetic recombinant adenovirus vector carrying the human growth and differentiation fac-tor-5(GDF-5) gene by using AdEasy-1 adenovirus vector system and to amplify and prepare the recombinant adenovirus in HEK 293 cells .Methods Human GDF-5 gene obtained by PCR was inserted into plasmid pMD19-T and the 1 .7 kb GDF-5 cDNA sequence was cloned into the adenoviral shuttle plasmid pShuttle-cytomegalovirus(CMV) of the AdEasy-1 system .It was identified by DNA sequencing and a digestion with Hind Ⅲ restriction enzyme .The resultant pShuttle-CMV-GDF-5 was used to generate the adenovi-ral GDF-5 vector through homologous recombination with the adenoviral backbone plasmid ,pAdEasy-1 in BJ5183 bacterial cells .It was selected by kanamycin and identified by a digestion with Hind Ⅲ restriction enzyme and amplified in XL10-Gold competent bac-teria .The DNA of recombinant adenovirus vector was finally linearized by Pac Ⅰ and the adenoviral recombinants were used to pro-duce adenoviruses in HEK293 packaging cells ,resulting in an Ad-GDF-5 identified by Western blot .The virus titer was assayed by TCID50 .Results GDF-5 cDNA sequence obtained by PCR was 1 .7 kb .Gene sequencing results indicated that the sequence was i-dentical with the one in GENBANK .Cloned sequence 1 .7 kb(GDF-5) was obtained by a digestion with Hind Ⅲ restriction enzyme after GDF-5 cDNA segment was cloned into pShuttle-CMV and AdEasy-1 .Western blot showed that two bands migrating at ap-proximately 15 and 55 kDa were clearly observed in PVDF membrane .These data confirmed that HEK293 cells expressed a large number of mature GDF-5 protein after infected with Ad-GDF-5 .Our research results demonstrated that recombinant adenovirus vector GDF-5 was successfully constructed .The virus titer was 5 .6 × 109 PFU/mL .Conclusion Recombinant adenovirus vector carrying the human GDF-5 gene is successfully constructed by using the AdEasy-1 adenovirus vector system .Moreover ,the Ad-GDF-5 with high titer is prepared .These provide the basis for further study of the biological function of GDF-5 and the gene thera-py of its related diseases .

OBJECTIVETo construct a eukaryotic expression plasmid pcDNA3.1(-)-Humanin.

METHODSThe recombinant plasmid pGEMEX-1-Humanin was digested with restriction endonucleases BamH I and Hind III and the Humanin gene fragments, about 100 bp length, were obtained. Then the Humanin gene fragments were inserted into eukaryotic expression vector pcDNA3.1(-) and the recombinant plasmids pcDNA3.1(-)-Humanin were identified by sequencing.

RESULTSRecombinant plasmid DNA successfully produced a band which had the same size as that of the Humanin positive control. The sequence of recombinant plasmids accorded with the Humnain gene sequence.

CONCLUSIONSA eukaryotic expression plasmid of Humanin was successfully constructed.

Base Sequence ; Cloning, Molecular ; methods ; Escherichia coli ; genetics ; metabolism ; Humans ; Intracellular Signaling Peptides and Proteins ; Molecular Sequence Data ; Molecular Weight ; Plasmids ; genetics ; Protein Engineering ; methods ; Proteins ; chemistry ; genetics ; metabolism ; Recombinant Proteins ; biosynthesis ; chemistry

Objective Hepatic artery (HA) reconstruction is one challenging procedure in pediatric liver transplantation (PLT).Here we review the first 115 microsurgical reconstructions of HA in PLT performed by a single surgeon,aiming to demonstrate the learning curve and the problems encountered.Methods From July 2016 to January 2017,a series of 115 microsurgical reconstructions of HA in PLT for end-stage liver disease were finished by one single surgeon with 4-year liver surgery experience and 2-week microsurgical training.HA reconstruction was performed with an operating microscope (Carl-Zeiss S88).Reconstruction was completed with interrupted sutures with 8-0 or 9-0 Prolene using the double clip for fixation.The blood flow was examined by Doppler scan daily after PLTs in first week and then once in 2nd week and first month for patency.A total of 143 artery anastomoses were performed in 115 PLTs.The age ranged from 3 months to 9 years.Indications for PLT included biliary atresia (105/115),Alagille syndrome (5/115),PFIC (3/115),Caroli disease (1/115),methylmalonicacidemia (1/115) and glycogen storage disease (1/115).Most of the PLTs were living donor liver transplantation (107/115),along with OLT (5/115) and split LT (3/115).Results The diameter of the arteries was mostly less than 2 mm (98/115).Up to date,one HA thrombosis (HAT) occurred at D8 after LT and 4 cases suspected as temporal HA stenosis (HAS) around 2 weeks after LT,which manifested as low velocity (<20 cm/s) and resistance index (<0.50) by Doppler.The HAT case failed in emergent re-anastomosis,but had a spontaneous recanalization at 3 weeks and is now in good condition without biliary problem.All the HAS children recovered to normal flows at first month.All children with HA complications started warfarin upon detection,with a targeted INR between 1.5-2.0.There were 6 deaths in this series including 5 cases of infections and 1 case of graft failure.Learning curve suggested a two phases growth (first 44 cases practicing phase vs.next 71 cases mature phase),which can be attributed to experience accumulation in terms of precise of manipulation,choice of inflow arteries for better match and stronger pulsation,avoidance of length redundant,prevention of kink.All the HAT and HASs happened in practicing phase while outcomes were excellent in mature phase.Moreover,time for each anastomosis was significantly shortened in second phase from 45-70 min to 30-55 min.Conclusion Microsurgical technique is highly safe in pediatric HA reconstruction,especially for very tiny arteries.It is possible to achieve low risk of complications for a new surgeon with adequate experience in liver surgery and microsurgical training.However,more surveillance and timing anticoagulation therapy is required before the mature of microsurgical technique.

The endophytic microorganisms found widely in many kinds of plants mediate various effects to theirs hosts. In this study, seven different dominant endophytes (SDE01 to 07) isolated from a Hy-peraccumulator-Solanum nigrum L. were resistant to Cd2+, and the strain SDE06 survived even in the medium containing 80 mg/L of Cd2+. Bacteria strain SDE06 was identified as Bacillus sp.. The removal of Cd2+ of SDE06 in different conditions were studied. Under the optimal conditions, the incubating time was 36 h, the solution pH 6.0, the temperature was 37?C and the Cd2+ concentration of medium was 20 mg/L, the highest removal rate was up to 80.2% at this condition.

18 fungal strains including N.coenophialum,N.lolii, N.huerfanum、N.chisosum、N.aotearoae、N.sp.and 8 varieties of grass seeds belonging to Festuca arundinacea and Lolium perenne have been studied.With amplification of IS1～IS3 and F1～R1 of genomic DNA, the primers Tub-2-F～Tub-2-R from Tubulin-2 gene and F3～R3 from NC25 gene have been designed.A PCR method to detect N.coenophialum and N.lolii was established, and also a nested-PCR method to detect N.coenophialum and N.lolii in single seed was established.These PCR detection methods are strongly special and much credible and rapid-speeded.