Child, Preschool ; Female ; Humans ; Infant ; Male ; Methionine ; blood ; Tyrosine ; blood ; Tyrosine Transaminase ; deficiency ; Tyrosinemias ; blood ; diagnosis ; enzymology ; pathology ; therapy

Objective:To evaluate the anti-HIV-1 activity of two new nonnucleoside reverse transcriptase inhibitors (NNRTIs), JB25 and JB26, in combination with 3 approved drugs (AZT, EFV, SQV)in vitro.Methods:The serially diluted 10 concentrations of JB25 and JB26 were combined with 7 serially diluted AZT, EFV and SQV respectively.The combination was added to 384 cell culture plates and then cocultured with HIV-1 ⅢB infected MT-2 cells for 3 days. Finally, the HIV-1 production was determined by measuring the expression of reporter genes of TZM bl cells. The data were analyzed by MacSynergy Ⅱ software.Results:The average capacity of synergism/antagonism of JB25 with AZT, EFV and SQV was 244.45/-5.05(nmol/L)~2%, 119.58/-65.93 (nmol/L)~2% and 145.83/-0.32 (nmol/L)~2% respectively;the average capacity of synergism/antagonism of JB26 with AZT, EFV and SQV was 398.90/0(nmol/L)~2%, 103.62/-0.49(nmol/L)~2% and 138.473/-0.27 (nmol/L)~2% respectively. Conclusion:Two new NNRTIs JB25 and JB26 develop synergism when combined with 3 approved drugs, respectively. MacSynergy Ⅱ software could evaluate the anti-HIV-1 activity of drug combination.

OBJECTIVETo clone, identify and phylogenetically characterize a clade B-Thai HIV isolate representing the most prevalent virus in Henan province.

METHODSPeripheral blood mononuclear cells (PBMCs) from an HIV-1 infected patient in Henan Province were separated, and co-cultivated with phytohemagglutinin-stimulated healthy donor PBMCs. Proviral DNA was extracted from productively infected PBMCs. The full-length HIV-1 genome was amplified by using the LA Tag long template PCR system. Primers were positioned in conserved regions within the HIV-1 long terminal repeats. Purified PCR products were T-A ligated into a pWSK29-T vector(CNHN 24 clone). Three recombinant clones containing virtually full-length HIV-1 genome were identified by PCR. The full-length genome was sequenced by using the primer-walking approach. Nucleotide sequence similarities were calculated by the local-homology algorithm. Phylogenetic trees of gag, pol and env reading frames were constructed using the Phylip software.

RESULTSHIV-1 C3V4 sequences indicate that the epidemic in this area was B-Thai subtype. V3 loop multiple amino acid sequence alignments showed amino acid alterations at nine positions. The 9,010 bp genomic sequence derived from isolate CNHN 24 contained all known structural and regulatory genes of an HIV-1 genome. No major deletions, insertions, or rearrangements were found. The highest homologies of the gag, pol, vpr, and vif reading frames to the corresponding clade B-Thai RL 42 sequences were 95.42%-97.08%. Phylogenetic trees showed the closest relationship of CNHN 24 and RL 42.

CONCLUSIONThe cloning and characterization of a virtually full-length HIV-1 B-Thai subtype in central China was completed in our laboratory. The data should be helpful to future studies on the genetic diversity of HIV-1.

Amino Acid Sequence ; Base Sequence ; Blood Donors ; China ; Cloning, Molecular ; DNA, Viral ; genetics ; Female ; Genome, Viral ; HIV Infections ; virology ; HIV-1 ; classification ; genetics ; Humans ; Leukocytes, Mononuclear ; virology ; Phylogeny ; Reading Frames ; Sequence Analysis, DNA ; Sequence Homology

No abstract available.
Irritable Bowel Syndrome*

OBJECTIVETo determine whether long-term treatment of attention deficit hyperactivity disorder (ADHD) with methylphenidate influences the growth in height and weight of children.

METHODSAnalyses were performed on 146 school age children (126 boys) diagnosed as ADHD and treated with methylphenidate [0.27-0.64 mg/(kg.day)] for methylphenidate group and 29 children with ADHD who did not receive any medication for ADHD (controls). These children were followed-up for 2-4 years. Changes in height and weight after long-term treatment with methylphenidate were recorded and the factors affecting growth of height, weight, and height velocity were analyzed.

RESULTSThe change of difference between patients' height and mean height in methylphenidate group and controls was (-1.86 +/- 0.82) cm (paired t test, t = 27.335, P < 0.001) and (-0.26 +/- 0.51) cm (P < 0.05), respectively; the change of height standard deviation score (SDS) in methylphenidate group and controls was -0.14 +/- 0.23 SD (paired t test, t = 7.326, P < 0.001) and +0.05 +/- 0.10 SD (P < 0.05), respectively. When the height change and height SDS change in methylphenidate group and controls were compared by using independent-samples T-test, the t value was -10.078 and -4.262 respectively, P for both was < 0.001. Both of bivariate correlation analysis and stepwise multiple-regression analysis indicated that the duration of treatment contributed significantly to the variance in change of height (P < 0.001); but age, sex, DSM-IV type, NJ22 degree and dose of methylphenidate did not contribute significantly to the variance of height. The mean height velocity from 1st to 4th year was 4.28 cm/year, 4.90 cm/year, 4.98 cm/year and 4.95 cm/year, respectively. With Friedman test, Chi-square = 253.673, P < 0.001. The change of difference of patients' weight to weight for height after methylphenidate was (-0.14 +/- 1.25) kg (paired t test, t = 1.326, P > 0.05).

CONCLUSIONSmall but significant deceleration of height velocity is the identified long-term side effect of methylphenidate, the magnitude of height deficit is related to duration of treatment. The height velocity was significantly attenuated in the first year. Methylphenidate had no significant influence on weight.

Attention Deficit Disorder with Hyperactivity ; drug therapy ; Body Height ; drug effects ; Body Weight ; drug effects ; Case-Control Studies ; Central Nervous System Stimulants ; adverse effects ; therapeutic use ; Child ; Child Development ; Female ; Humans ; Male ; Methylphenidate ; adverse effects ; therapeutic use ; Regression Analysis

Objective To determine whether morphine having the ability to influence the antiviral effect of lamivudine(3TC)in vitro study.Methods MT2 cells were randomly assigned into morphine+3TC treatment group,morphine+naloxone+3TC treatment group,naloxone+3TC treatment group.Both 3TC and virus control groups were set up.The corresponding MT2 cells were treated with opiates antagonist(naloxone)for 0.5 hours before the 24-hours morphine treatment program was implemented while all of the groups were then infected with equal amounts of cell-free HIV-1 ⅢB strain and 3TC.HIV-1 p24 antigen in culture supernatants collected at days 3,4,5 and 6after infection status was tested and the inhibition of 3TC anti-HIV-1 p24 antigen of various treatment groups calculated.Results Inhibition of 3TC anti-HIV-1 p24 antigen of Morphine+3TC treatment group was the lowest when HIV-1 infected cells at 3rd and 4th day and showed significant difierence (P<0.05)when compared to the 3TC control.However,there was no statistically significant difference among them(P>0.05),when virus was infected the cells at 5th and 6th day.The difference of 3TC anti-HIV-1 p24 antigen inhibition between the morphine+naloxone+3TC treatment group and the naloxone+3TC treatment group was not significant(P>0.05).Similar results were obtained when these two groups were compared to the 3TC control group(P>0.05),respectively.The 3TC anti-HIV-1 p24 antigen inhibition of each treatment group reduced as the time of infection prolonged,showing a significant and time-course effbct.Conclusion The 3TC antiviral effect was reduced by morphine in the early stage of infection,and could be blocked by naloxone.

Objective:To investigate the subtype distribution of HIV-1 strains prevalent in four areas of China,and to study the characteristics of gag gene variation and changes in antigen epitopes under the host immune pressures. Methods:The plasma of HIV-1 infected people from Henan, Guangdong, Sichuan and Beijing in China were collected. Virion RNA was extracted directly from plasma after the virion was condensed. The gag gene was amplified by RT-PCR and nested-PCR.Sequences were subtyped by Genotyping Tool software, and phylogenetic analysis of gag gene were performed using the MEGA 4.1 software.The gene distances intra each subtype were calculated by Distance program. The Ks/Ka ratios were calculated using SNAP program. The variation analysis of CTL antigen epitopes restricted by main HLA-Ⅰ specificities in China was performed.Results:Six subtypes or circulating recombinant forms(CRFs)of HIV-1,including B',CRF07_BC,CRF01_AE,B,CRF08_BC and CRF02_AG,were identified in four areas of China.The gene distances intra each subtype were CRF01_AE>B>CRF08_BC> CRF07_BC>B' listed in order of size, meanwhile the order of Ks/Ka ratios was CRF01_AE>B>CRF08_BC>B'>CRF07_BC. Far more diversity of antigen epitopes in P17 region was observed than that in P24.Epitope mutations intra subtypes were CRF01_AE>B>B'>CRF07_BC listed in order of size. Conclusion:Itseems that CRF01_AE is under the strongest immune pressures,and displays the most diversity of gene and variation of epitopes intra subtypes prevalent in China, followed by subtype B, B' and CRF07_BC. The discrepancy of epitope mutations intra the subtypes is significant.

OBJECTIVETo study the applied valuation of Onodera prognostic nutrition index (Onodera index) in elderly patients with colorectal cancer.

METHODSOnodera indexes of 163 elderly patients with colorectal cancer were calculated and these patients were divided into better-nourished group (Onodera index ≥45) and under-nourished group (Onodera index <45). Correlations of Onodera index with general data, operation type, postoperative complication, recovery of gastrointestinal function, clinicopathological feature and prognosis were analyzed. Cox proportional hazards model was also established to identify the independent prognostic factors for prognosis of elderly patients with colorectal cancer.

RESULTSPatients in better-nourished group had significantly higher radical resection rate [90.9% (70/77) vs. 62.8% (54/86), P<0.01], lower postoperative complication rate [17.1% (12/70) vs. 53.7% (29/54), P<0.01] and earlier postoperative defecation [(3.09±1.14) d vs. (3.43±1.98) d, P<0.05] than those in under-nourished group. Onodera index was found to be related to age, tumor location, tumor size, and operation type (all P<0.05). Better-nourished group had significantly better survival than worse-nourished group (5-year survival rate: 64% vs. 24%, P<0.01). Onodera index was identified as an independent prognostic factor for elderly patients with colorectal cancer (RR=0.888, 95%CI:0.800-0.985, P=0.025).

CONCLUSIONOnodera index is a valuable clinical marker in preoperative estimation as well as prognosis prediction for elderly patients with colorectal cancer.

Aged ; Aged, 80 and over ; Colorectal Neoplasms ; surgery ; Female ; Humans ; Male ; Nutrition Assessment ; Prognosis ; Retrospective Studies

OBJECTIVETo study the role of PCR technique in detection of mycobacterium tuberculosis in the samples from joint tuberculosis, and to evaluate the clinical value of PCR in diagnosis of joint tuberculosis.

METHODSFrom June 1993 to August 2001, PCR was used to detect DNA of mycobacterium tuberculosis, and the standard culture was applied to detect mycobacterium tuberculosis. Mycobacterium tuberculosis were respectively blindly by the two techniques in the samples obtained from 95 patients with joint tuberculosis (55 males and 40 females, the age ranging from 2 to 75 years, with an average of 34 years). The positive rate of mycobacterium tuberculosis detection was calculated.

RESULTSIn the detection of mycobacterium tuberculosis, positive rate was 82% (78/95) in PCR technique, and 16% (15/95) in standard culture technique. There were statistical differences between the two groups (chi2=67, P<0.001). The whole process of PCR amplification was automatic and could be finished within several hours, and the detecting time was considerably shorter.

CONCLUSIONPCR technique is a rapid, simple, sensitive and specific method for detection of mycobacterium tuberculosis in the samples of joint tuberculosis, showing more marked advantages than the standard culture technique. It is valuable in the early rapid diagnosis and differential diagnosis of joint tuberculosis.

Adolescent ; Aged ; Child ; Child, Preschool ; Culture Techniques ; methods ; Female ; Humans ; Male ; Middle Aged ; Mycobacterium tuberculosis ; genetics ; isolation & purification ; Polymerase Chain Reaction ; methods ; Tuberculosis, Osteoarticular ; diagnosis ; microbiology ; Young Adult

OBJECTIVETo investigate the factors of the human immunodeficiency virus (HIV) associated with sexual-transmission in central China.

METHODS(1) Cross-sectional study: couples that one was HIV positive were selected in Henan and Hebei province of China. The couples must be 20 - 50 years old with normal function on sexual intercourse. Cordant couples that subsequently infected partners were at risk of infection solely through sexual contact with the HIV-seropositive partner and the discordant couples that the seronegative partners were at risk of infection solely through sexual contact with the HIV-seropositive partner, were selected. Plasma viral load, CD4 cell count were tested. (2) Case-control study was used to compare 7 sexual transmitted cases and 56 nontransmitted controls with respect to the frequency of sexual intercourse, plasma viral load and CD4 cell count.

RESULTS(1) A total of 87 couples that at least one partner was HIV positive were recruited include 56 discordant couples and 7 cordant couples with whom sexual transmission had happened. The rate of sexual transmission was 11.1% among those at the risk of sexual transmission. (2) Of the discordant couples, male positive rate 25%, female positive was 75%. (3) The risk for transmission was higher in those couples with the frequency of unprotected vaginal sexual intercourse (> or = 4 times per month) than the reference group (< 4 times per month) (Fisher's exact test, P = 0.047, OR = 8.0). Median plasma viral load was significantly higher in the antecedent infected partners of cordant couples than the positive partner of discordant couples (378,285.71 vs 136,578.57 copies/ml, t = 3.591, P < 0.01). The odds ratio was 22.0 for plasma viral load > or = 100,000 copies/ml compared with the reference group of < 100 000 copies/ml (Fisher's exact test, P = 0.016). The CD4 cell count and CD4/CD8 of the transmitted group were significantly lower than that of the nontransmitted (t = 2.767, P < 0.05; t = 6.06, P < 0.05).

CONCLUSIONSThe frequencies of heterosexual-transmission in central China were relatively low. The risk of heterosexual transmission was related to the frequency of sexual intercourse. Higher plasma viral load and lower CD4 count was strongly correlated with high risk of heterosexual transmission.

Adult ; Blotting, Western ; CD4 Lymphocyte Count ; China ; Coitus ; Cross-Sectional Studies ; Disease Transmission, Infectious ; Female ; HIV ; immunology ; HIV Infections ; transmission ; Heterosexuality ; Humans ; Male ; Middle Aged ; Risk Factors ; Spouses ; Viral Load