In order to study the pathogenesis of HPVl1 and seek for a therapeutic approach of the disease caused by HPV1l. The HPVl1/E2 644bp was amplified by PCR with HPVll plasmid DNA. pGEM-T Easy was used as vector and a clone pTV-644 was obtained.The inserted DNA sequencing was carried out after selecting and identification. According to the hammerhead structure described by Symon's, the possible secondery cleavage sites on HPVll/E2 mRNA were analyzed and the secondery structure of substrate was predicted by computer and ribozyme, excluding analogous sequence of substrate combined with ribozyme were found in the mRNA.The hammerhead ribozyme of RZ2777 against HPV1l/E2 mRNA was seleted to carry out cleavage reaction in vitro. Results of the experiment showed that 644bp substrate derived from HPVll/E2 can be cleavaed site-specifically by ribozyme in vitro. The cleavage activity showed over 85% by choosing the optimun reaction condition, which was not affected by two cis-ribozymes on both 3′-and 5′-ends released by self cleavage,but two flan-king sequences of target RNA influenced the cleavage activity. Results demonstrated that the ribozyme will become a highly effective and specific therapy against HPVll infection.

Objective To observe the immunomodulatory effects of homologous bone marrow mesenchymal stem cells (MSCs) obtained from the bone marrow in rats after heart transplantation. Methods Twenty adult male Lewis rats were used as donors for the heart transplantation, whereas twenty adult male Wistar rats served as recipients. The recipients with cervical heart transplantation were randomly divided into two groups (10 each). Approximately 3ml 0.9% NaCl solution was injected through the tail vein 24h after heart transplantation in the control group (group A). About 2×106 MSCs (suspended in 3ml 0.9% NaCl solution) were injected through the tail vein 24h after heart transplantation in the MSCs treatment group (group B). Four recipient rats from each group were randomly chosen one week after transplantation for determining proportion of CD4+ T, CD8+ T, CD4+CD25high T, and CD4++CD25highfoxp3+ T cells in the lymphocytes in the venous blood and grafts. Subsequently, the CD4+/CD8+ ratio was calculated. The survival time of the grafts were observed in the remaining six rats in each group. Results (1)The survival time of the transplanted hearts was 7.2±1.3d in group A, and 14+.8±2.9d in group B, showing a significant difference between the two groups (P<0.01). (2)The ratios of CD4+/CD8+, CD4+CD25high T cells/total lymphocytes and CD4+CD25highFoxp3+ T cells/total lymphocytes in venous blood were 1.18±0.07, 2.51%±0.56% and 2.05%±0.62% respectively in group B, which were higher than those in group A (0.4+9±0.05, 0.96%±0.19%, 0.82%±0.09%). These results showed significant statistical differences (P<0.01). (3)The ratios of CD4+/CD8+ in the allografts showed no statistical difference between the two groups (P>0.05). The ratios of CD4+CD25high T cells/total lymphocytes and CD4+CD25highFoxp3+ T cells/ total lymphocytes in the allografts were evidently higher in group B (2.74+%±0.28%, 2.54+%±0.31%) than in group A (0.61%±0.06%, 0.53%±0.06%), showing a significant statistical difference (P<0.01). Conclusion Intravenous infusion with MSCs from the bone marrow of the recipients can induce immune tolerance and prolong the survival time of transplanted heart in rats.

Objective To investigate the change of the basic fibroblast growth factor(bFGF) leve in human detrusor muscle(DM)in bladder outlet obstruction(BOO)due to benign prostatic hyperplasia(BPH)and its implication.Methods Fifty-four patients with BPH were divided into two groups:the obstructive DM stability and instability groups;and 15 men with bladder tumor who underwent operation in the same period were enrolled in the control group.The bFGF mRNA level in DM was measured by reverse transcription and polymerase chain reaction(RT-PCR)and the bFGF protein level was measured by immunohistochemical staining method.Results The bFGF-mRNA expression level of bladder smooth muscle cells was significantly lower in the control group than that in the obstructive DM stability and instability groups(all P＜0.05),but there was no significant difference between the obstructive DM stability and instability groups(P＞0.05). Conclusions The expression level of bFGF mRNA in bladder DM is elevated in BOO due to BPH,but there is little or no correlation between the increased expression of bFGF mRNA and detrusor muscle instability.

OBJECTIVETo observe the preventive protection of Kangdu Bufei Decoction (KBD) on acute severe respiratory syndrome (SARS) in medical personnel of Hong Kong at the epidemic period.

METHODSAt the epidemic period of SARS in Hong Kong, there were 2,601 medical staffs administered with KBD distributed by the Center. It was confirmed that 1,063 persons had taken it for successive two weeks according to the request and returned the effective questionnaire, they were regarded as the TCM group. The control group was consisted of 15,374 subjects who didn't take Chinese herbal medicine. Contents of the questionnaire including the condition of testee in terms of quality of life (QOL), changes of influenza-like symptoms and Warm disease symptoms. Serum immunological examination had been conducted in 37 of the persons in the TCM group.

RESULTSNo one in the TCM group got SARS infection while in the control group, 64 (0.4%) had got. Self-control before and after treatment examination showed that significant improvement appeared in the TCM in aspects of influenza-like and Warm disease symptoms and QOL. The results of serum immunological examination showed that after administration of KBD, the immunity of organism was improved and elevated.

CONCLUSIONKBD could prevent the occurrence of SARS, it is possibly realized through improving symptoms, elevating QOL and enhancing immunity of organism.

Adjuvants, Immunologic ; therapeutic use ; Adult ; Cross Infection ; prevention & control ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Hong Kong ; Humans ; Infectious Disease Transmission, Patient-to-Professional ; prevention & control ; Male ; Middle Aged ; Phytotherapy ; Severe Acute Respiratory Syndrome ; prevention & control ; transmission ; Surveys and Questionnaires

The main purpose of this study was to investigate the active components of the Chinese medicine formula Shenqi San (SS) by high performance liquid chromatography with diode array detector and electrospray ionization-hybrid quadrupole time-of-flight mass spectrum (HPLC-DAD-ESI-QTOF-MS),and demonstrate the anticancer mechanism of SS on human lung adenocarcinoma A549 cells by evaluating the cell proliferation and apoptosis induction.The chloroform extraction of SS (CE-SS) was extracted from SS,while HPLC-DAD-ESI-QTOF-MS assay was performed to identify components of CE-SS.MTT assay was used to quantify the proliferation of A549 cells with the treatment of CE-SS.Apoptosis analysis was carried out by detecting phosphatidylserine (PS) externalization using the Annexin V-FITC Apoptosis Detection Kit and the stained cells were analyzed with a flow cytometer.DAPI staining assay was carried out to observe morphological characteristics of apoptotic cells.Western blotting was used to detect the expression of important signaling proteins including caspase-3,-8,-9,p53,Bax and Bcl-2.Eight compounds were identified through HPLC-DAD-ESI-QTOF-MS analysis and 3-pyridine carboxylic acid,barbatin C,scutebarbatine F and barbatine D might be the main compounds responsible for the antitumor effect of CE-SS.CE-SS suppressed the proliferation of lung cancer A549 cells in a time-and dose-dependent manner.By Annexin V-FITC/PI double staining,we found that treatment with CE-SS induced apoptosis in A549 cells.After 24-h exposure to CE-SS,the expression of cleaved-caspase-9,cleaved-caspase-8 and cleaved-caspase-3 protein was activated,the expression of p53 protein increased while the ratio of Bax/Bcl-2 also increased.This study identified the eight compounds of CE-SS,and demonstrated their anticancer effect on human lung adenocarcinoma A549 cells via induction of apoptosis.

OBJECTIVE: To study genetic polymorphism of 7 Y-specific short tandem repeats (STR) and assess their usefulness in forensic casework. METHODS: 7 Y-STR have been amplified in two multiplex reactions, (Multiplex I:DYS391, GATA-A4, GATA-A10 and GATA-H4. Multiplex II:DYS439,DYS437 and DYS434). PCR products were separated by polyacrylamide gels electrophoresis followed by silver stain. RESULTS: When 372 unrelated individuals from the Han population in Guangdong were detected by those system, DYS391, GATA-A4, GATA-A10, GATA-H4,DYS439,DYS437 and DYS434 showed 5,7,6,5,6,4,4 alleles, respectively. A total of 254 different haplotypes were identified, of which 201 (79.13%) were found in single individuals. The overall haplotypes diversity reached 0.9960. CONCLUSION The 7 Y-STR loci are highly genetic polymorphism and they will be very powerful for establishing Y-STR database, understanding human origin, paternity testing and personal identification.
Alleles ; Asian People ; Chromosomes, Human, Y/genetics* ; Forensic Medicine ; Gene Frequency ; Genetic Markers ; Genetics, Population ; Genotype ; Haplotypes ; Humans ; Male ; Microsatellite Repeats/genetics* ; Paternity ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Sequence Analysis, DNA

OBJECTIVE: To establish a multiplexing Y-STR system and study haplotype frequencies of 4 Y-specific loci in China Han population. METHODS: DYS439, DYS390, GATA-A7.2 and DYS393 loci were amplified simultaneously and were analyzed by polyacrylamide gel electrophoresis and silver staining. RESULTS: When 558 unrelated male individuals from the Han population in China were tested by the multiplex system, DYS439, DYS390, GATA-A7.2 and DYS393 show 7,7,7,6 alleles, respectively. 180 different haplotypes were detected. The power of discrimination of this system was 0.9853. CONCLUSION The multiplex amplified system of these 4 Y-specific loci and their database are useful for human origin exploration and forensic practice.
Alleles ; China ; Chromosomes, Human, Y ; DNA/isolation & purification* ; Electrophoresis, Polyacrylamide Gel ; Ethnicity/genetics* ; Female ; Forensic Medicine ; Gene Frequency ; Haplotypes ; Humans ; Male ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Silver Staining ; Tandem Repeat Sequences/genetics*

OBJECTIVETo detect the single nucleotide polymorphism (SNP) of chemokine regulated upon activation, normal T cell expressed and secreted (RANTES) promoter and first intron of asymptomatic, human immunodeficiency virus 1 (HIV-1) infected individuals of in Han Chinese and evaluate the influence on HIV-1 infection by variants.

METHODSCase-control study was adopted, Genotypes of RANTES promoter -403 and -28 from 538 samples and RANTES first intron In1.1 from 300 samples of Han Chinese were detected by DNA sequencing or by polymerase chain reaction-restriction fragment length polymorphism.

RESULTSThere were six genotypes of RANTES promoter -403 and -28 found in Han Chinese group. The distribution of genotypes was AC/AC 12.4%, AC/AG 3.5%, AC/GC 29.2%, AG/GC 10.9%, GC/GC 42.1%, AG/AG 1.5%. The haplotypes was GC 62.7%, AC 28.7%, AG 8.6%. Compared with AC/AC, Odd ratio (OR) of RANTES genotypes AC/AG, AC/GC, AG/GC, GC/GC was associated with weaker reduced susceptibility to HIV-1 infection. However, there were no significant contents of the allele frequencies between people living with HIV-1 and healthy individuals. The distribution of RANTES In1.1 alleles were T/T 71.0%, C/T 19.9%, C/C 9.1% and haplotypes were RANTES In1.1T 81%, In1.1C 19%, respectively; There were significant differences of RANTES In1.1 between people with HIV-1 infection and healthy individuals in males. The In1.1C-bearing genotypes would increase susceptibility to HIV-1 infection but no significant differences in females were found. Strong linkage disequilibrium was observed between all of the three RANTES SNPs.

CONCLUSIONThe two -403A/G, -28C/G variants in RANTES promoter region and intron In1.1 T/C mutation genotype were found to be associated with the genetic susceptibility to HIV-1 infection among the Han Chinese. However, the In1.1C allele or its haplotypes in RANTES intron 1 displayed a stronger dominant association with HIV-1 infection in males.

Adolescent ; Adult ; Aged ; Case-Control Studies ; Chemokine CCL5 ; genetics ; metabolism ; Child ; China ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; genetics ; Genotype ; HIV Infections ; genetics ; HIV-1 ; Haplotypes ; Humans ; Introns ; genetics ; Male ; Middle Aged ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length ; Polymorphism, Single Nucleotide ; Promoter Regions, Genetic ; genetics ; Sequence Analysis, DNA ; Sex Factors

Objective To discuss the clinical effect of the ramified musculocutaneous flap pedicled with the descending branch of lateral circumflex femoral artery treatment of bones and arthrsis of extremity in- fections with soft tissue defects.Methods The muscle flap blooded with the muscular branch of lateral fem- oral muscle and musculocutaneous flap blooded with the musculocutaneous branch were designed,all of which were pedicled with the descending branch of lateral circumflex femoral arteryIn clinic24 cases of bones and arthrosis of extremity infections with soft tissue defects were treated with this kind of ramified musculocutaneous flap.Results Of the 24 cases23 cases were survived while 1 case was lost16 cases were healed at stageⅠ8 cases were healed at stageⅡSinus has formated in 3 casesone of which twicebut they were healed in one year with the treatment of debridmentsFour cases with osteomvelitis and bone defect were treated with bone grafting in the later 6～8 months after the wound has healedTwenty-two cases were followed-up for 6～20 monthsinfcetiou didn't recur.Conclusion This kind of ramified musculocutaneous flap has such ad- vantages as longer blood vessel pediclefilling the defects completely flexible application and stronger anti-in- fectionthat it may be an effective way in treating bones and arthrosises of extremity infections with soft tissue defects.

Aged ; Animals ; China ; epidemiology ; Disease Outbreaks ; Female ; Humans ; Male ; Middle Aged ; Orientia tsutsugamushi ; Scrub Typhus ; epidemiology