The baculovirus expression system has been used extensively for the expression of recombinant proteins in insect cells.Recently,recombinant baculoviruses containing mammalian cell-active promoter element have been used to transduce a broad spectrum of primary and established mammalian cells,including non-hepatic cells.Recombinant baculoviruses have been used successfully for transient or stable gene delivery in mammalian cells in vitro,while the efficiency of delivering gene in vivo is inhibited obviously by complements,but efforts have been made to overcome the problems,for instance,VSV-G-pseudotyped baculoviruses display complement resistance.The mechanism of the transduction is still not clearly understood,though many researches have been done.The baculovirus is able to replicate only in insect cells,but it is incapable of initiating replication cycle in mammalian cells,which guarantees high biosafety of this gene delivery system.In addition,this system is easily manipulated and able to carry large inserts.These attributes will undoubtedly lead to the increased application and continued development of this system for highly effective gene delivery into mammalian cells.

Objective To assess the safety and efficacy of the local nerve block anesthesia procedure in daysurgery inguinal herniorrhaphy practice.Methods Fifty-eight inguinal herniorrhaphy were performed under the local nerve block anesthesia in the form of day surgery between April 2014 and April 2015 other 58 inguinal hemiorrhaphy were performed under general anesthesia.Time of operation,hospitalization stay,complications and costs were evaluated.Results The average operating time of the day-surgery group was (47 ± 12) min.All operations were completed successfully.Fifty-six patients were discharged in 24 hours and 2 patients were discharged in 48 hours after the surgery.The average costs were 8437 yuan.The rate of postoperative complications was approximately 6.6％ (4/58),including hematoma(3 cases),urinary relention(1 case),and chronic pain (1 case).The VAS scores of 6 hours after the operation were (2.4 ± 0.8).Conclusions It suggests that preperitoneal hemiorrhaphy under the local nerve block anesthesia in the form of day-surgery was safe and effective.Also,it could significantly shorten the time of hospitalization and reduce medical costs.

Treatment based on syndrome differentiation is an essential feature of traditional Chinese medical diagnosis. The interventions based on changes of syndrome types in randomized controlled trials are complicated, leading to the difficulty of blind method enforcement. This article described a double-blind method. It could be used in randomized controlled trials under the condition of different syndrome types and different medications. It numbered drugs in two stages, and in two phases to achieve double-blind. This method not only guaranteed investigators and subjects to be in blinded conditions, but also achieved using different medications for patients of different syndromes. It also caused no drug waste. It was scientific and feasible.
Double-Blind Method ; Humans ; Medicine, Chinese Traditional ; Randomized Controlled Trials as Topic ; Single-Blind Method

OBJECTIVE:To observe therapeutic efficacy and safety of Danshen chuanxiongqin injection combined with flunari-zine hydrochloride in the benign prevention and treatment of paroxysmal positional vertigo (BPPV) and lower extremity deep ve-nous thrombosis (DVT) in post-operative long-term bedridden patients with lower limb fractures. METHODS:300 post-operative long-term bedridden patients with lower limb fractures were selected and randomly divided into observation group and control group,with 150 cases in each group. Control group was given Flunarizine hydrochloride capsules orally 10 mg,qd;observation group was additionally given Danshen chuanxiongqin injection 10 ml+5% Glucose injection 250 ml,ivgtt,qd. The incidence of BPPV and DVT were observed in 2 groups after intervention,and the circumference of lower limb,blood coagulation indexes, blood rheology indexes and inflammatory factor were observed before and after intervention,and the incidence of ADR was com-pared. RESULTS:The incidence of BPPV and DVT in observation group were 18.0% and 16.7%,which were significantly lower than in control group(48.7% and 52.7%),with statistical significance(P<0.05);after intervention,the circumference of lower limb,blood rheology indexes and the levels of inflammatory factors in 2 groups were decreased significantly, while the coagula-tion indicators were significantly improved;the observation group was better than the control group,with statistical significance (P<0.05). There was no statistical significance in the incidence of ADR between 2 groups(P>0.05). CONCLUSIONS:Danshen chuanxiongqin injection combined with flunarizine hydrochloride is effective in the prevention of BPPV and DVT in long-term bed-ridden patients with lower limb fractures,with low incidence of ADR.

Aim To study relative bioavailablity of cefaclor effervescent tablets in healthy volunteers. Methods According to the crossover design, A volunteers were each orally given a single does of the 0.75 g cefaclor effervescent tablets and cefaclor capsules with an interval of 5 days between the two formulations.The plasma concentrations of the drug were determined by RP-HPLC.Pharmacokinetic parameters were obtained by ATPK programe,and calculated on the basis of open single compartment model.Results After a single oral dose, the peak levels in plasma averaged Cmax(31.27?5.81)?g?ml-1 and(30.56?5.25) ?g?ml-1 at (0.58?0.12)h and(0.73?0.17)h and AUC0～4(35.48?4.65) ?g?h?ml-1 and (35.89?2.90) ?g?h?ml-1 for tablet and capsule,respectively. Conclusion The result shows that two formulations are bioequivalence.

Multiple epitopes from one or more viruses can be lined up and co-expressed in one vector to generate multi-epitopes DNA vaccines. In the study, four recombinant plasmids were constructed based on HA and NP gene of avian influenza virus (AIV) (H5N1): (1) pIRES/HA, carrying the complete HA gene; (2) pIRES/tHA, carrying a truncated HA gene fragment of major neutralizing antigenic epitopes; (3) pIRES/tHA-NPep, in which three CTL epitopes of NP gene of AIV were fused to the truncated HA from the C-terminal; and (4) pIRES/tHA-NPep-IFN-gamma, which was constructed by replacing neo gene in pIRES/ tHA-NPep with IFN-y of chicken. Fifty five SPF chickens were randomly divided into five groups and immunized with the above four constructs and control plasmid. Each chicken was intramuscally immunized with 200 microg plasmid DNA three times in a two-week interval. Two weeks after the third immunization, chickens were injected with H5N1 subtype avian influenza virus. Before the virus loading no detectable antibodies to HA were found in the chicken serum; but high levels of HI antibodies were detected in the serum of the survived chickens. The percentages of CD4+ and CD8+ T lymphocyte in peripheral blood of immunized chickens increased steadily after the vaccination. After virus loading all chickens in the control group died within three to eight days, and the survival rates of the four DNA vaccine groups were as follows: pIRES/HA, 54.5%; pIRES/tHA, 30%, pIRES/ tHA-NPep, 36.3%, pIRES/tHA-NPep-IFN-gamma, 50%. These results indicated that multi-epitopes DNA immunization can induce immune response and protect chickens from homologous virus loading.
Animals ; Chickens ; Epitopes ; immunology ; Influenza A Virus, H5N1 Subtype ; immunology ; pathogenicity ; Influenza in Birds ; immunology ; prevention & control ; virology ; Vaccines, DNA ; immunology

Objective To explore the methods of maintaining the stability of middle turbinate (MT),reducing the incidence of middle meatus synechia formation and improving the treatment effect of endoscopic sinus surgery.Methods A prospective study was conducted in 54 consecutive patients with chronic sinusitits.The patients were allocated into two groups.During the operation,the most anterior and superior MT attachments to the agger nasi region were preserved,and anterior and posterior ethmoidectomy were performed in group A(27 cases 47 sides).Besides above structures,the most posterior and inferior aspects of the basal lamellae and the horizontal boney strut structures of the basal lamellae were preserved in group B(27 cases 51 sides).Results There were no significant differences between the two groups in age,course and preoperation visual analogue scale(VAS)score.The VAS scores in group A before and 1 year after operation were(6.41±0.25)and(1.70±0.36),the difference was significant(t=10.472,P<0.05).The VAS scores in group B before and 1 year after operation were(6.78 ±0.23)and(0.66 ±0.16),the difference was significant(t:17.195,P<0.05).The Lund-Kennedy scores in group A and group B one year after operation were(1.85±0.47)and(0.67±0.16),the difference was significant (t=2.290,P<0.05).The MT position was described as stable,sligilt drifting laterally and synechia formation.And the incidenee of stable,slight drifting laterally and synechia between MT and the nasal lateral wall in group A and group B was 57.4%,23.4%,19.1% and 88.2%,3.9%,7.8% respectively,the differences were significant(X~2=12.511,P<0.05)Conclusions Conservation of the horizontal bony strut of basal lamellae could better maintain the stability of MT,reduce the incidence of MT drifting laterally and synechia formation,and finally improve the curative effect of endoscopic sinus surgery.

In order to investigate the biological effects of the VP22?-mE6?/mE7 built-in adjuvant fusion protein vaccine on the tumor associated with HPV-16 infection.HSV-1 VP22? and HPV-16 mE6?/mE7 genes were cloned,and the pET28a-VP22?-mE6?/mE7 recombinat prokaryotic expression vector was constructed.Vector was transformed into Rosetta(DE3)E.coli string and expressed under the induction of IPTG.The re-naturalized protein was then purified via Ni2+ affinity adsorbent column and identified by SDS-PAGE and Western blot.Purified protein was immunized BalB/C and C57BL/6 mice to evaluate the immunogenicity and anti-tumor activity.The expressed recombinant protein formed as inclusion body with a prediction MW about 34kDa and contained approximately 45% of total somatic protein.The VP22?-mE6?/mE7 immunization induced higher titer of specific IgG against HPV,higher level of lymphocyte proliferation and better effect on suppressing HPV16 positive TC-1 tumor growth than the mice immunized with mE6?/mE7 alone.The results showed that the recombined built-in adjuvant vaccine could induce specific cellular immune response in vitro and inhibit the TC-1 tumor proliferation in vivo,that would be a foundation for further studies and developments of inner adjuvant vaccines.

We have shown previously that a Semliki Forest virus (SFV) replicon vectored DNA vaccine (pSFV1CS-E2) expressing the E2 glycoprotein of classical swine fever virus (CSFV) conferred full protection for pigs immunized three times with 600 microg of the vaccine. This study aims to evaluate the efficacy of the DNA vaccine with lower dosage and fewer inoculations. Pigs were immunized twice with 100 microg pSFV1CS-E2 (n = 5) or control plasmid pSFV1CS (n = 3), respectively. Pigs immunized with pSFV1CS-E2 developed high titers of specific neutralizing antibodies against CSFV after the booster, and the antibody titers increased rapidly upon challenge. The immunized animals showed no clinical symptoms except short-term fever and low-level viremia, whereas the control pigs immunized with the control plasmid produced no detectable antibody before challenge and showed obvious clinical signs following challenge, and 2 pigs died on 10 or 11 days post-challenge. All control animals developed extended viremia as detected by nested RT-PCR and real-time RT-PCR. Severe pathologic lesions typical of CSFV infection were observed at necropsy. We conclude that the alphavirus replicon-vectored DNA-based vaccine can be potential marker vaccine against CSFV.
Animals ; Antibodies, Neutralizing ; blood ; immunology ; Antibodies, Viral ; blood ; immunology ; Body Temperature ; immunology ; Classical Swine Fever ; blood ; immunology ; prevention & control ; Classical swine fever virus ; genetics ; immunology ; Genetic Vectors ; genetics ; Immunization ; Plasmids ; genetics ; Replicon ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Semliki forest virus ; genetics ; Swine ; virology ; Time Factors ; Vaccines, DNA ; administration & dosage ; genetics ; immunology ; Viral Envelope Proteins ; genetics ; immunology ; Viremia ; genetics ; immunology

OBJECTIVETo investigate the effect of hyperbaric oxygen(HBO)therapy on mitochondrial free radicals after transient focal cerebral ischemia in rats.

METHODSThe male SD rats were randomly assigned into two groups, control and HBO groups. All animals were subjected to 90 min intra-luminal middle cerebral artery occlusion (MCAO) with the regional cerebral blood flow monitored in vivo by laser Doppler flowmetry. HBO treatment was performed in a pressure chamber with 100% O(2)(3 ATM 1 h) 3 h after ischemia. Twenty-four hours after ischemia, mitochondria in the ischemic core and penumbra were isolated and the contents of H(2)O(2), O(2)(*-), MDA, SOD, GSH-PX and GSH in mitochondria were measured respectively.

RESULTAfter cerebral ischemia-reperfusion, contents of mitochondrial H(2)O(2), O(2)(*-), MDA increased, while the SOD, GSH-PX and GSH in the mitochondria decreased significantly both in the ischemic core and the ischemic penumbra, compared with those in the normal controls(P<0.05). In the ischemic penumbra, HBO therapy increased significantly the content of O(2)(*-)(P<0.05), enhanced the activity of SOD, and decreased the level of MDA (P<0.05). However, HBO therapy did not change the level of MDA, though it also increased the content of O(2)(*-) and the activity of SOD in the ischemic core. HBO therapy had no significant effect on the contents of H(2)O(2), GSH-PX and GSH in the ischemic mitochondria.

CONCLUSIONHBO therapy initiated early after acute transient cerebral ischemia in rats can increase the mitochondrial free radicals level, but also increase the activity of the anti-radical enzymes. HBO treatment inhibits the lipid peroxidation damage of mitochondria in the ischemic penumbra, but not in the ischemic core, which indicates that the mitochondrial function plays a role in the reaction of the free radical in the ischemic area after HBO therapy.

Animals ; Free Radicals ; metabolism ; Glutathione Peroxidase ; metabolism ; Hyperbaric Oxygenation ; methods ; Infarction, Middle Cerebral Artery ; metabolism ; therapy ; Ischemic Attack, Transient ; metabolism ; therapy ; Male ; Mitochondria ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; prevention & control ; Superoxide Dismutase ; metabolism