Objective To provide experimental evidences for developing a safe and effective re- combinant fowlpox virus which can prevent the infection of HIV-2.Methods A fowlpox virus(FPV) transferring vector was constructed by inserting HIV-2 gag gene to the downstream of a synthetic complex promoter ATI-p7.5?20 of vector pUTA2.Transfection was then carried out,and recombi- nant FPV(rFPV)was screened by 5'-bromo-deoxyuridine(BrdU),genome PCR and western blot detection.Balb/c mice were immunized with rFPV by muscular injection.Anti-HIV-2 antibody, CD4~+ and CD8~+ T-cell count and specific target-killing activity of spleen CTL in immunized mice were analyzed by ELISA,FACS and LDH release assay,respectively.Results A transferring vector pA- gag was constructed and confirmed by amplifying a fragment of 766 bp from the rFPV genome.Mean- while,HIV-2 multi-antibody-specific protein blot(55 000)was detected from the recombinant virus and the HIV-2 specific antibody was detected from the immunized Balb/c mice.HIV-2 specific target- killing activity of spleen CTL was observed in immunized mice.Conclusion A recombinant fowlpox virus expressing HIV-2 structural protein Gag has been obtained,and it can stimulate HIV-2-specific eelluar and humoral immune reactions in mice.

OBJECTIVETo explore the effect of moxibustion at Sanyinjiao (SP 6) on normal process of childbirth and safety of mothers and infants.

METHODSOne hundred and seventy-four cases of singleton primiparas, fetal head position were selected as the subjects and divided into three groups by single-blind randomized method: an acupoint group (n = 59), a non-acupoint group (n = 57) and a blank group (n = 58). When the cervix was dilated by 2 to 3 cm, DAJ-23 type multi-function moxibustion apparatus was used at Sanyinjiao (SP 6) in acupoint group, and at non-acupoint spot (the midpoint from the radial arterial pulse throbbing place on the radial side of the transverse crease of the wrist to the radial end of biceps tendon of the elbow) in non-acupoint group, 30 min for each one. The conventional labor care was adopted in blank group. The differences were observed in the first active stage, the second stage and the third stage of the labor as well as the safety of mothers and infants among three groups.

RESULTSThe durations of the first active stage of labor [(123.05 +/- 96.401) min] and the second stage [(37.407 +/- 22.390) min] in acupoint group were shorter than those [(171.40 +/- 94.339) min, (49.517 +/- 25.373) min] in non-acupoint group, as well as those [(178.79 +/- 97.231) min, (53.684 +/- 20.087) min] in blank group separately (all P < 0.05). It was discovered that the progressions of the first active stage and the second stage of labor were accelerated in acupoint group, and there was no statistically significant difference in the third stage of labor among the three groups (all P > 0.05). Bleeding amount after childbirth in acupoint group was less than those in non-acupoint group and blank group (both P < 0.05), and Apgar scores of newborns in acupoint group and non-acupoint group were higher than that in blank group (both P < 0.05).

CONCLUSIONMoxibustion at Sanyinjiao (SP 6) can shorten the process of childbirth, reduce postpartum bleeding and has no harmful impact on mothers and infants.

Acupuncture Points ; Adult ; Female ; Humans ; Labor, Obstetric ; Moxibustion ; Pregnancy ; Young Adult

BACKGROUND: Greatly importance has been attached to ceramic-on-ceramic bearing surface due to its excel ent wear resistance. But the risks of squeaking and ceramic fracture also go with it. Up til now, the choice between ceramic-on-ceramic and ceramic-on-polyethylene bearing surfaces in primary total hip arthroplasty remains controversial. OBJECTIVE: To compare the clinical outcomes and safety between ceramic-on-ceramic versus ceramic-on-polyethylene bearing surfaces in total hip arthroplasty based on meta analysis. METHODS: We electronical y searched databases including PubMed/Medline, Embase, Web of Science, Cochrane Col aboration database, Chinese Biomedical Literature Database (CBMdisc) and China National Knowledge Internet for randomized control ed trials on the comparison between ceramic-on-ceramic versus ceramic-on-polyethylene bearing surfaces in total hip arthroplasty from inception to January 2015. References of the included studies were also retrieved. Investigators severely selected the studies, extracted data and assessed the quality according to the inclusion and exclusion criteria. Then, meta-analysis was performed using RevMan 5.2 software. RESULTS AND CONCLUSION: Nine randomized control ed trials were included, involving 1 231 hips with ceramic-on-ceramic prosthesis and 932 hips with ceramic-on-polyethylene prosthesis. Meta analysis showed that both bearing surfaces achieved satisfied function recovery. But ceramic-on-ceramic had significantly increased risks of squeaking and ceramic fracture, meanwhile ceramic-on-polyethylene showed significantly higher wear rate. There were no significant differences in intra- or post-operative dislocation, osteolysis and other complications and prosthesis failure with any reason between two bearing surfaces. These results suggest that during the short- to mid-term fol ow-up period, no sufficient evidence can tel that ceramic-on-ceramic was obviously super than ceramic-on-polyethylene. Long-term fol ow-up is required for further evaluation.

OBJECTIVETo study the role of the HBV-infected mothers' PBMC in intrauterine transmission of HBV to their fetuses.

METHODSThirty pregnant women with serum HBV DNA negative and PBMC HBV DNA positive and their newborns were used as the study group. Ten pregnant women with serum HBV negative and their infants served as the control group. HBV DNA in serum and in PBMC was detected using nested polymerase chain reaction (n-PCR). The mothers' PBMC in newborns' peripheral blood was examined using heminested-PCR.

RESULTSFour newborns were serum HBV DNA positive and 8 newborns were HBV DNA positive in PBMC in the study group. Among them, 2 newborns were HBV DNA positive in both serum and PBMC, 6 cases were positive in PBMC only, and 2 cases were positive in serum only. Five mothers had the GSTM1 gene; and it was not detected in 3 newborns. Among the 8 newborns with HBV DNA positive in PBMC, 3 did not have the GSTM1 gene, at the same time their mothers possessed the GSTM1 gene. Mothers' PBMC were detected in all of these three newborns' peripheral blood. HBV DNA in serum and in PBMC of the control group infants were all negative.

CONCLUSIONHBV-infected PBMC of the mother may serve as a vector in HBV intrauterine infection.

Adult ; DNA, Viral ; analysis ; Female ; Hepatitis B virus ; isolation & purification ; Hepatitis B, Chronic ; blood ; transmission ; Humans ; Infant, Newborn ; Infectious Disease Transmission, Vertical ; Leukocytes, Mononuclear ; virology ; Pregnancy ; Pregnancy Complications, Infectious ; virology

OBJECTIVETo study the mechanism and significance of peripheral blood mononuclear cell (PBMC) of neonates infected with hepatitis B virus (HBV).

METHODSEighty-four HBsAg-positive and HBeAg-negative mothers and their newborns were recruited in this study. Sixteen hepatitis B virus markers (HBVM)-negative mothers and their neonates were served as control. All these cases had no symptoms of hepatitis, serious pregnancy complications and preexisting disease. Age, gestational age and the method of delivery were matched in two groups (P > 0.05). Five ml blood samples were taken from the peripheral vein of the pregnant women before delivery and from neonates within 24 hours after birth, before inoculation of HBV vaccine (HBVac). Serum and PBMC were isolated from 2 ml and 3 ml samples respectively. The sera, PBMC and the last supernatant of PBMC washing were stored at -80 degrees C. HBVM of neonates were detected by using enzyme linked immunosorbent assay (ELISA). HBV DNA in serum, PBMC and the last supernatant of PBMC washing of mothers and neonates were detected by using a nested-polymerase chain reaction (n-PCR). Two pairs of oligonucleotide primers, the outer primer pair for first PCR and inner primer pair for second PCR, designed according to region S of HBV genome were synthesized at Shanghai Cell Biology Institute of Chinese Academy of Sciences. The neonates who were HBV DNA positive in PBMC but HBsAg and HBV DNA negative in serum were followed up for one year, HBsAb in serum and HBV DNA in PBMC were observed in the neonates.

RESULTS(1) The positive rate of HBV DNA in 84 serum and PBMC of mothers were 53.57% and 40.48%, respectively (chi(2) = 2.891, P > 0.05). All the results were weakly positive. (2) Twenty-four (28.57%) newborns in the study group were infected, including 7 who were only HBV DNA positive in serum, 11 only HBV DNA positive in PBMC and 6 in both, all the results were weakly positive. HBsAg was negative in all the newborns. None of the neonates in control group was infected with HBV. There was significant difference between the two groups (chi(2) = 4.55, P < 0.05). (3) Of all the study cases, 11 (13.10%) neonates were HBV DNA weakly positive in PBMC but HBsAg and HBV DNA negative in serum. Of their mothers, 5 were only HBV DNA positive in serum, 2 only positive in PBMC and 4 positive in both serum and PBMC. Seven of the 11 neonates were followed up for one year and at the end of follow-up, 4 were HBsAb positive and HBV DNA negative in PBMC; 3 were HBsAb negative, and among the 3 cases HBV DNA in 2 was still positive in PBMC, HBsAg and HBV DNA in serum were negative in all the 7 neonates.

CONCLUSION(1) HBV DNA positivity either in serum or in PBMC in mothers can result in infection of PBMC with HBV in their neonates. (2) PBMC infection with HBV can exist for a long time in neonates while HBsAg and HBV DNA are negative in serum, and may result in vaccination failure in neonates.

Case-Control Studies ; DNA, Viral ; blood ; Female ; Hepatitis B ; diagnosis ; immunology ; Hepatitis B Vaccines ; administration & dosage ; Hepatitis B virus ; Humans ; Infant, Newborn ; Infectious Disease Transmission, Vertical ; Leukocytes, Mononuclear ; virology ; Pregnancy

To screening out Chinese vaccine candidate against HIV-1, Chinese vaccine strain 282E4 of fowlpox virus was used as the vector to construct the recombinant fowlpox virus (rFPV) coexpressing gp120 of Chinese HIV-1 strain and IL-18, and the recombinant virus was indentified by PCR and Western blot. The specific DNA fragment could be amplified by PCR from the genome of rFPV. Western blot analysis showed that gp120 and IL-18 could be expressed not only in chicken embryo fibroblast (CEF) cells infected by rFPV, but also in mammalian cells infected by rFPV. After the recombinant fowlpox virus was inoculated into BALB/c mice, the spleen specific CTL activities and serum antibodies in the immunized mice were detected, which demonstrated that the rFPV had good immunogenicity and could induce BALB/c mice to produce specific humoral and cellular immunity. IL-18 palyed the role of immunoadjuvant. The study lays the basis on the preparation of genetic engineering live vector vaccine against HIV-1.
AIDS Vaccines ; immunology ; Adjuvants, Immunologic ; Animals ; Antibodies, Viral ; immunology ; Fowlpox virus ; genetics ; immunology ; metabolism ; Gene Transfer Techniques ; Genetic Vectors ; HIV Envelope Protein gp120 ; biosynthesis ; genetics ; immunology ; HIV-1 ; genetics ; immunology ; Humans ; Immunization ; Interleukin-18 ; biosynthesis ; genetics ; immunology ; Mice ; Mice, Inbred BALB C ; Vaccines, DNA ; immunology

OBJECTIVETo explore the mechanism of acupoint sticking of "Hua yutie" in improving ischemic stroke.

METHODSEighty rats were randomly divided into 5 groups, a model group, an acupoint sticking group, an acupuncture group, a Nimodipine group and a normal group. Middle cerebral artery occlusion (MCAO) was used for preparation of focal cerebral ischemic rat model. After modeling, any treatment was not given to the model group; for the acupoint sticking group, "Hua yutie" was applied at "Dazhui" (GV 14) ,"Qihai" (CV 6) and "Mingmen" (GV 4); for the acupuncture group, acupuncture was given at the same acupoints as those in the acupoint sticking group; the Nimodipine group received intragastric administration of Nimodipine. And the normal group did not receive any treatment. Their infarction volume, the cerebral water content, expression of vascular endothelial growth factor (VEGF) and the protein level were observed.

RESULTSThe infarction volume coincided with the dominative scope of the middle cerebral artery of the electric coagulation. There were significant differences in the cerebral water content as the various treatment groups compared with that of the model group (all P<0.05). The VEGF positive cell number and the protein level around the infarction area in the acupoint sticking group were increased as compared with those in the model group (P<0.01), with no significant difference as compared with the Nimodipine group and the acupuncture group (all P>0.05).

CONCLUSIONAcupoint sticking of "Hua yutie" alleviates the cerebral damage after ischemia possibly through enhancing the expression and protein level of VEGF.

Acupuncture Points ; Administration, Cutaneous ; Animals ; Brain Ischemia ; drug therapy ; genetics ; metabolism ; Cerebral Infarction ; drug therapy ; genetics ; metabolism ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Gene Expression ; drug effects ; Humans ; Male ; Rats ; Rats, Wistar ; Vascular Endothelial Growth Factor A ; genetics ; metabolism

Objective To develop a method for the determination of total cholesterol in serum by isotope dilution liquid chromatography tandem mass spectrometry (ID-LC/MS/MS).Methods Serum samples were supplemented by addition of [3,4-~(13)C_2]-cholesterol,hydrolyzed with alcoholic sodium hydroxide and oxidized into cholest-4-ene-3,6-dione by chromic acid.The oxidation products were analyzed by LC/MS/MS using atmospheric pressure chemical ionization (APCI) source and detection modes of multiple reaction monitoring (MRM) and single ion recording (SIR).Signals (peak areas) of the internal standard were corrected for the contributions of cholesterol and the signal ratios of cholesterol to internal standard for the calibrations were linearly regressed against cholesterol concentrations.The resulted regression equation was used for the calculation of serum cholesterol concentrations.Results The correlation coefficients between the peak area ratios and cholesterol concentrations were 0.999 9 and higher.Under MRM mode,the average within-run CV of the results obtained on 3 serum samples was 0.95% (ranged from 0.92% to 0.99%) and the total CVwas 0.86% (0.82% to 0.89%),and under SIR mode,the within-run CV was 0.64% (from 0.54% to 0.77%) and the total CVwas O.69% (0.62% to 0.81%),respectively. Results on certified reference materials (SRM 1951 a Level Ⅰ and Level Ⅱ;GBW 09145 and GBW 09147) showed an average bias of 0.23% (0.14% to 1.00%) under MRM mode,and 0.24% (0.07% to 1.27%) under SIR mode.Conclusions An ID-LC/MS/MS method for serum cholesterol has been developed.It is specific and precise and may be used as a candidate reference method.

Objective To evaluate the enhanced magnification endoscopy in the diagnosis of Barrett esophagus,and to explore the relationship between mucosal surface patterns and pathological epithelial types of Barrett esophagus.Methods Enhanced magnification endoscopy was performed 'after spraying 2%-3% acetic acid on the surface of distal esophagus in 40 Barrett esophagus patients.Mucosal specimen were biop- syed.Results According to the mucosal types of Toyoda in 2003,there were three mucosal types:Ⅰ dot pat- tern 7(17.5%),5 of 7(71.4%)fundie type,Ⅱ reticular pattern 24(60.0%),16 of 24(66.7%)fundic type,Ⅲ cerebroid/villous 9(22.5%),intestinal metaplasia or dysplasia.Conclusion Enhanced magnifi- cation endoscopy helps to identify areas with intestinal metaplasia and dysplasia,and is useful in the diagno- sis of Barrett esophagus.

OBJECTIVETo compare antiproliferation effects of vinblastine nanopraticles and vinblastine water solution in human glioma cell lines BT325.

METHODVinblastine nanoparticles were prepared by emulsion polymerization process and using dextran as a stabilizing agent. It was characterized by means of morphology, size, drug entrapment efficiency and loading efficiency. Human glioma cell lines BT325 were treated with different concentrations of vinblastine nanoparticles and vinblastine water solution for 48 h, Antiproliferation effect was measured by MTT method. Morphological changes were observed by inverted microscope, transmission electron microscope and scanning electron microscope.

RESULTMean diameter of VLB-PBCA-NP was about 74.4 nm, and drug entrapment efficiency and loading efficiency was 78.47% and 39.24%, respectively. Cell growth inhibition rate of vinblastine nanoparticles group and vinblastine water solution group in a concentration range (5-5 000 g x L(-1)) for 48 h was 41%, 49%, 73%, 83% and 28%, 33%, 54%, 60% respectively. Entrapment of VLB in NPS may distinctly degrade absorbency as compared to free drugs. Glioma cell BT325 which treated with VLB water solution were initial stage of apoptosis, and apoptosis body were forming. But VLB NPS-treated BT325 cells were intermediate or end stage, and missed structure integrality.

CONCLUSIONVLB-PBCA-NP and VLB water solution could inhibit the growth of human glioma cell lines BT325, and VLB nanoparticles have stronger inhibition effect compared with VLB water solution in the same dose. PBCA may be effective as promising carrier for the transport of vinblastine into the glioma cells.

Antineoplastic Agents, Phytogenic ; pharmacology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Microscopy, Electron, Transmission ; Nanoparticles ; Vinblastine ; pharmacology