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.

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.

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

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

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.

Adult ; HIV Infections ; complications ; diagnosis ; Humans ; Lymphohistiocytosis, Hemophagocytic ; diagnosis ; etiology ; Male

OBJECTIVETo investigate the role of endotoxin receptor expression in the activation of hepatic stellate cells (HSCs).

METHODSHSCs were isolated from normal rats and the expression of endotoxin receptors on quiet HSCs and in vitro activated HSCs was determined using RT-PCR and immunocytochemical staining methods. A rat model of liver fibrosis and cirrhosis was established. The expressions of CD14 and alpha-SMA in liver tissues were detected by immunohistochemical staining.

RESULTSFreshly isolated HSCs had a low level of CD14 mRNA expression and no expression of TLR4 mRNA was detected. The in vitro activated HSCs had increased expressions of CD14 mRNA and TLR4 mRNA and LPS up-regulated the expression of endotoxin receptors. Immunocytochemical staining showed cytoplasmic and nucleolus staining for CD14 in the cultured HSCs. LPS played a further role on CD14 protein expression. In the development of liver fibrosis, the number of CD14-positive cells in the livers was increased and these cells were distributed along the sinusoids. In the later stage of liver fibrosis, the CD14-positive cells were gathered in the fibrotic septae, which also contained alpha-SMA positive cells.

CONCLUSIONThe activated HSCs expressed endotoxin receptors. The endotoxin receptors may be involved in the role in which HSCs played in the inflammatory process and liver fibrosis development.

Actins ; metabolism ; Animals ; Cells, Cultured ; Hepatic Stellate Cells ; metabolism ; Lipopolysaccharide Receptors ; metabolism ; Liver Cirrhosis ; metabolism ; pathology ; RNA, Messenger ; genetics ; Rats ; Rats, Wistar ; Receptors, Immunologic ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo investigate the effect of ginsenoside-Rg3 on lung metastasis of ribonuclease inhibitor (RI) gene-transfected mouse B16 melanoma.

METHODSC57BL/6 mice were iv injected with parental or RI-transfected B16 melanoma cells. Lung metastasis was assessed by the number of surface tumor nodules. Mice were divided into 6 groups. Group I, II and III of mice were given parental, mock-transfected and RI-transfected B16 melanoma cells, respectively while in group IV, V and VI, Rg3 (1.5 mg/kg, iv q.o.d. x 10) was given to mice bearing parental, mock-transfected and RI-transfected B16 melanoma, respectively. Micovessel density (MVD) of the lung metastatic tumor was assessed by immunohistochemical staining of factor VIII-R expression.

RESULTSThe number of tumor nodules was significantly decreased in mice injected with RI-transfected B16 melanoma (Gp III, compared to Gp I and II). Rg3 treatment per se could also decrease the number of lung tumor nodules but to a lesser extent (Gp IV and V compared to Gp III). However, Rg3 synergized with RI transfection resulting in most significant inhibition of lung metastasis (Gp VI). Mice in Gp I and II died within 26 days of the experiment, whereas all the mice in Gp VI were alive during the observation period of one and one half month. MVD was significantly decreased in the lung tumor nodules in mice injected with RI-transfected B16 melanoma. It was further decreased when additional Rg3 was given (Gp VI).

CONCLUSIONTransfection of ribonuclease inhibitor gene significantly reduces the metastatic potential of B16 melanoma. Ginsenoside-Rg3 has a synergistic effect.

Animals ; Antineoplastic Agents, Phytogenic ; pharmacology ; Cell Line, Tumor ; Ginsenosides ; isolation & purification ; pharmacology ; Lung Neoplasms ; pathology ; secondary ; Male ; Melanoma, Experimental ; genetics ; pathology ; Mice ; Mice, Inbred C57BL ; Neoplasm Transplantation ; Panax ; chemistry ; Placental Hormones ; genetics ; Transfection