Objective To investigate the therapeutic effect of large dose mucosolvan combined variant flow rate continuous positive airway by nasal mask on neonatal respiratory distress syndrome(NRDS).Methods One hundred and fourteen newborns with NRDS were randomly divided into treatment group(58 cases) and control group(56 cases),on the base of same combined therapy,the cases in control group only underwent oxygen-absorbing by head set with the flow rate 4-6 L/min,and the cases in treatment group were given large dose mucosolvan(Ambroxol Hydrochloride) 30 mg/(kg?d) + 5%GS 20 mL,for two times and variant flow rate continuous positive airway by nasal mask(NCPAP),the parameter setting flow rate 6-8 L/min,FiO_2 0.4-0.6,pressure 5-8 cm H_2O.The clinical symptom and blood gas analysis after 12 and 48 hours were observed and compared the changes of pa(O_2),pa(CO_2),pa(O_2)/FiO_2 in two groups.Results The dyspnea and groan in 44 cases in the treatment group lessoned or vanished,pa(O_2) rised and pa(CO_2) lowered,the oxygenation index obviously increased,the cases with RDS grade Ⅰand gradeⅡ had better therapeutic effect,and the cases with RDS grade Ⅲ(X-ray)and Ⅳ had not manifest effect,the total effective rate was 75.8% in treatment group and 26.7% in control group.There were significant difference in therapeutic effect and oxygenation index between two groups.Conclusions Large dose mucosolvan(combi)-ning variant flow rate continuous positive airway by nasal mask can significantly improve the ventilation and oxygenation function and there are significant therapeutic effect in NRDS,especially in the NRDS grade Ⅰand gradeⅡ,the trachea cannula may be avoided and mechanical ventilation rate may be decreased if the therapeutic method can be used in earlier period.

China ; epidemiology ; Dengue ; epidemiology ; Dengue Virus ; Humans

OBJECTIVETo establish a liquid chip technology to detect Y chromosome microdeletions in Chinese infertile males with azoospermia or oligozoospermia.

METHODSMultiplex PCR and liquid chip technology were used to detect the Y chromosome microdeletions in AZF region in 178 infertile patients with azoospermia and 134 infertile patients with oligozoospermia as well as 40 fertile control men.

RESULTSForty out of 312 patients (12.8%) were found to have deletions in AZF region. The microdeletion frequency was 14%(25/178) in the azoospermic group, 9.6%(11/114) in the oligospermic and 20%(4/20) in the severe oligospermic group.

CONCLUSIONThe authors developed a high-throughput, fast and simple assay to screen the AZF region microdeletions of Y chromosome.

Asian Continental Ancestry Group ; genetics ; Azoospermia ; genetics ; Base Sequence ; Chromosome Deletion ; Chromosomes, Human, Y ; genetics ; Electrophoresis, Polyacrylamide Gel ; Humans ; Male ; Oligonucleotide Array Sequence Analysis ; methods ; Oligospermia ; genetics ; Sequence Tagged Sites

OBJECTIVETo construct a recombinant lentiviral vector (pXZ208-BDDhFVIII) mediating B-domain-deleted human coagulation factor VIII (BDDhFVIII) gene and investigate its expression in HLF, Chang-Liver and MSC cells.

METHODSBDDhFVIII gene fragment was separated by endonuclease digestion and was cloned into the multiple cloning sites of pXZ208 to construct a recombinant lentiviral vector pXZ208-BDDhFVIII. Viral particles were prepared by means of three-plasmid cotransfection of 293T package cells by calcium phosphate precipitation. After infection, the coagulant activity of human FVIII in the culture medium of 293T, HLF, Chang-Liver and MSC cells was assayed by one-stage method. The gene transduction efficiency was assayed by flow cytometry (FCM). Furthermore, PCR was performed to test the integration of BDDhFVIII.

RESULTSThe infection rates of HLF, Chang-Liver and MSC were (74.52 +/- 7.57)%, (27.24 +/- 6.53)% and (42.34 +/- 5.84)% respectively. The activities of FVIII in supernatants of HLF, Chang-Liver and MSC were (54.1 +/- 5.6)%, (22.5 +/- 2.9)% and (12.5 +/- 2.7)% respectively. BDDhFVIII gene integration was detected in all the infected cells.

CONCLUSIONThe recombinant lentiviral vector pXZ208-BDDhFVIII was successfully constructed and efficiently integrated into target cells to express human FVIII activity in vitro.

Cell Line ; Factor VIII ; biosynthesis ; genetics ; metabolism ; Gene Expression ; Genetic Vectors ; Humans ; Lentivirus ; genetics ; Plasmids ; Transfection

To investigate the influence of Anxin granules combined with tirofiban on acute myocardial infarction (AMI) Patients after elective percutaneous coronary intervention (PCI). One hundred and twenty AMI patients were randomly divided into treatment group and control group. The patients in the two groups were all given Tirofiban 30mins before PCI . The treatment group was added Anxin granules 30 mins before and after PCI. Tissue factor (TF) and von willebrand factor (vWF) were tested at 6 hours after operation. Syndromatology alteration of traditional Chinese medicine (TCM) and bleeding complications were observed at 4 weeks after operation. Both TF and vWF at 6 hours after operation of the treatment group was lower than the control group significantly (P < 0.01), while the condition of myocardial ischemia at 90 mins after operation of the treatment group was better than control group with significance. The syndromatology alteration of TCM especially spontaneous perspiration and hypodynamia of the treatment group were improved significantly compared to control group 4 weeks after operation. All patients in both groups had no bleeding complications and thrombopenia. The study suggests that Anxin granules combined with tirofiba can improve the clinical efficacy and the endothelial function of AMI patients after PCI with no increase in bleeding events.
Aged ; Angioplasty, Balloon, Coronary ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Humans ; Middle Aged ; Myocardial Infarction ; complications ; metabolism ; surgery ; Postoperative Hemorrhage ; drug therapy ; etiology ; metabolism ; prevention & control ; Thromboplastin ; metabolism ; von Willebrand Factor ; metabolism

OBJECTIVETo evaluate the development and operation on measles surveillance system.

METHODSTo formulate surveillance program, and then establish surveillance system on measles in Hainan province before the case surveillance was conducted. Descriptive epidemiological method was used to analyze the measles cases reported through the notifiable diseases epidemic situation report system and measles surveillance system during 2004 - 2008 in Hainan province.

RESULTSFrom 2004 to 2008, through surveillance system, 3040 suspected measles cases were reported which was 96.00% higher than that from the notifiable diseases epidemic situation report system. When the measles surveillance system was first established in 2004, the number of cases reported by the surveillance system was less than that from the epidemic report system, however, the sensitivity of the surveillance system had been increasing since 2005. In 2008, the number of suspected cases reported by surveillance system was 19.12 times more than from the epidemic report system. The proportion of confirmed cases in the total suspected cases was decreasing annually, from 90.34% to 4.48%, along with the increase of the sample collection rate, from 22.73% to 99.12%. The proportion of laboratory confirmed cases in the total confirmed cases increased from 7.55% to 86.97%. With suspected cases, the IgM antibody positive rate on measles and rubella were 31.57% and 34.52%, respectively. In Hainan, the epidemic pattern of measles had been sporadic, except for 2006 as there occurred several outbreaks in that year. The of disease incidence had an obvious seasonal peaks, from March to June. Measles mainly attacked children under 15 years of age, accounted for 86.68% of the age group. Children with no immunity or unknown immune history accounted for 85.67%.

CONCLUSIONThe sensitivity of surveillance system had been increasing annually since it was developed and the incidence had been dropping to its lowest level. These achievements had built a solid foundation for the eventual elimination of measles.

China ; epidemiology ; Disease Outbreaks ; prevention & control ; Humans ; Incidence ; Measles ; epidemiology ; prevention & control ; Population Surveillance

Objective To evaluate the application of western blot in treponema pallidum antibody screening among blood donors.Methods Through standard blood liquidation of syphilis,the sensitivity and specificity of two kinds of TP -ELISA regents and one TPPA regent were evaluated.TP -WB was used to test 279 positive specimens with TP -ELISA,and the correlation between test results of the two methods and the distribution characteristics of WB bands were analyzed. Results The sensitivity of the two kinds of TP -ELISA reagent was 1 00.00%,while the specificity were 92.86% and 85.71 % respectively.At the same,the sensitivity of TPPA reagent was 88.46%,while the specificity was 1 00.00%.In 279 positive specimens with TP -ELISA method,WB confirmed positive was 21 6,positive rate was 77.42%;Including S /Co value >5 and two kinds of ELISA reagent testing both positive specimens were 205,the WB confirmation test positive rate was 1 00.00%,accounted for 99.91 % in 21 6 WB positive samples.Orderly Logistic regression analysis,the method of ELISA S /Co value >5 and double reagent is positive,had statistical correlation with test positive for WB,respectively(P <0.01 );21 6 TP -WB positive specimens WB banding distribution analysis,TP1 7 belt with syphilis antibody IgM,TP1 5 belts and IgG +IgMand IgG exist statistical correlation respectively(P <0.01 ).Conclusion ELISA method of double reagent positive and S /Co value >5 specimens of the basic can be diagnosed with syphilis,WB test with positive results for blood donation member state judgment has guiding significance.

This study was aimed to construct a lentiviral vector carrying human coagulant factor VIII (FVIII) and to investigate its expression in 293T cells. B-domain-deleted factor VIII gene fragment (BDDhFVIIIcDNA) was obtained by enzyme digestion and cloned into lentiviral vector pXZ208 to establish the expression vector pXZ208-BDDhFVIII. Recombinant viral particles were prepared by cotransfection with packaging plasmid delta NRF and envelope plasmid VSV-G using calcium phosphate precipitation method. 293T cells were transfected by viral supernatant. Coagulant activity of FVIII, BDDhFVIIImRNA and genome integration were assayed by one-step method, RT-PCR and PCR after transfection. The results showed that 293T cells could be transfected by recombinant virus. The transfection rate of 293T was 59.57%. After transfection, the cells expressed FVIII efficiently. Detection confirmed that the activity of FVIII was 12%, 43% and 87% respectively at 24, 48 and 72 hours after infection. BDDhFVIII transcription was detected by RT-PCR from the infected cells. The gene integration in the targeted cells was also observed. It is concluded that the successfully constructed lentiviral vector is able to generate high level expression of human FVIII in 293T cells, which may provide a potential application of gene therapy to haemophilia A.
Cell Line ; Factor VIII ; genetics ; metabolism ; Gene Expression ; Genetic Vectors ; Humans ; Lentivirus ; genetics ; metabolism ; Recombinant Proteins ; genetics ; metabolism

The study was purposed to prepare the recombinant lentiviral vector pTK161 and pTK162 carrying B-domain-deleted canine factor (BDDcFVIII) gene, and to investigate whether the canine FVIII (cVIII) can be expressed in vitro. The BDDcFVIII gene was ligated behind PUB and 2OH1 promotors to create lentiviral vectors pTK161 and pTK162. Meantime lentiviral vectors pTK161' and pTK161' were produced by cloning a green fluorescent protein (GFP) into pTK151 and pTK152, which was driven by PUB and 2OH1 promotors respectively. Vector supernatant were prepared by using transfer calcium phosphate mediated-cotransfection of 293T cells. The virus vector, DeltaNRF packaging-plasmid, and VSV-G envelope-plasmid was assayed by titers and cFVIII activity in cell culture supernatant after infection into 293T cells. pTK161, pTK162, pTK161' and pTK161' were identified by restriction enzyme analyzing. The results showed that the lentiviral vectors pTK161, pTK162, pTK161' and pTK161' were successfully constructed, and the titers of pTK161' and pTK161' reached to 1.54 x 10(6) U/ml and 2.83 x 10(6) U/ml; the activity of cFVIII could be detected at 24 hours after infection of 293T cells by pTK161 and pTK162, and achieved the highest level at 72 hours later. The higher level of cFVIII activity was achieved by transfected with pTK162 than that of pTK161 (p < 0.05), which closed to the cFVIII activity in normal dog plasma. 1/4 of the highest level could be detected 6 weeks later. It is concluded that the prepared HIV1-based lentiviral vectors can infect 293T cells to express cFVIII effectively. The results provide the basis for further studying HIV-1-based lentiviral vector gene therapy for hemophilia A.
Animals ; Dogs ; Factor VIII ; biosynthesis ; genetics ; Genetic Vectors ; genetics ; HIV-1 ; genetics ; metabolism ; Recombinant Proteins ; biosynthesis ; genetics

This study was purposed to constructe the three-plasmid system of the lentiviral vector carrying the green fluorescent protein (GFP) gene and to investigate the expression of GFP in T lymphocytes of the mouse. The polypurine tract (PPT) element, ubiquinone promoter (PUB) and GFP were ligated to plasmid pLO134 using subcloning technology to construct plasmid pTK153. Human kidney 293T cells were co-transfected with the three-plasmid system containing packaging plasmid DeltaNRF, plasmid pTK153 and envelope plasmid VSV-G by using calcium phosphate DNA precipation and the expression of GFP was observed under fluorescence microscope after 12 hours. The viral particles were collected after transfection 72 hours, were frozen at -80 degrees C and were used to infect mouse T lymphocytes at multiplicity of infection (m.o.i.) of 3. The expression of GFP in mouse T lymphocytes was observed by fluorescence microscopy and fluorescence-activated cell sorting (FACS). The results showed that the transfection efficacy was 63.04 +/- 7.24% in 293T cells analysed by FACS and the viral titer was (3.09 +/- 0.61) x 10(6) U/ml. The expression of GFP was also evident in mouse T lymphocytes and the transduction efficacy was (37.98 +/- 6.26)%. It is concluded that the three-plasmid system of lentiviral vector containing GFP gene is successfully constructed and the transduction efficacy is high in mouse T lymphocytes.
Animals ; Genetic Vectors ; genetics ; Green Fluorescent Proteins ; biosynthesis ; genetics ; Lentivirus ; genetics ; Mice ; Mice, Inbred BALB C ; RNA, Viral ; analysis ; T-Lymphocytes ; metabolism ; Transduction, Genetic