OBJECTIVETo provide reference for the safe needling depth of Dazhui (GV 14), Jianzhongshu (SI 15), Xuanshu (GV 5) and Mingmen (GV 4) in clinical acupuncture and moxibustion treatment.

METHODSThirty-two adult volunteers were divided into 3 groups, thin person group, moderate person group and fat person group according to Luo's indexes, and computer-aided tomography was used to measure the needling depth of Dazhui (GV 14), Jianzhongshu (SI 15), Xuanshu (GV 5) and Mingmen (GV 4).

RESULTSThe safe depths of perpendicular needling were different for persons of different somatotypes, for example the needling depth for Dazhui (GV 14) was (32.86 +/- 3.96) mm for the thin person group, (37.76 +/- 4.91) mm for the moderate person group, and (47.93 +/- 5.30) mm for the fat person group.

Acupuncture Points ; Acupuncture Therapy ; Combined Modality Therapy ; Humans ; Moxibustion ; Tomography, X-Ray Computed

Three compounds were isolated from the extract of Taxus cuspidta Sibe et Zucc with the column chromatography on silica gel and preparative HPLC methods. Their structures were identified according to the physicochemical properties and spectral analysis, and they were identified as (E)-1-methoxy-2-O-(p-coumaroyl)-myo-inositol (1), 2-deacetoxy-7beta, 9a, 10beta-trideacetyltaxinine J (2) and (3aS, 4aR, 6S, 8S, 8aS, 9R, 10R, 10aS)-benz[f]azulene-6, 8, 9, 10 (3H)-terol, 3a, 4, 4a, 5, 6, 7, 8, 8a, 9, 10-decahydro-10a-(1-hydroxyl-1-methylethyl)-1, 8a-dimethyl-5-methylene (3). Among them, compound 1 was a new compound, and compounds 2, 3 were two novel natural products.
Azulenes ; chemistry ; isolation & purification ; Coumarins ; chemistry ; isolation & purification ; Inositol ; analogs & derivatives ; chemistry ; isolation & purification ; Molecular Structure ; Plants, Medicinal ; chemistry ; Taxoids ; chemistry ; isolation & purification ; Taxus ; chemistry

The aim of this study was to investigate the effects of tyrosine kinase inhibitor PTK787 on cell proliferation, cell cycle and the expression of fak mRNA of human chronic myeloid leukemia (CML) cell line K562, and to explore the mechanism of PTK787 against acute myeloid leukemia. The MTT method was used to detect the effects of PTK787 in various concentrations and at different time points on proliferation of K562 cells; the flow cytometry was used to determine the effects of PTK787 in different concentrations on cell cycle of K562 cells; the RT-PCR was used to assay the expression of fak mRNA in K562 cells treated with PTK787 for 48 hours. The results showed that along with increasing of the concentration and prolonging of time, the inhibitory rate of PTK787 on K562 proliferation was gradually enhanced. The comparison between various concentration groups at same time or comparison between various time groups in same concentration showed significant differences (p < 0.05), in which the effect of 320 micromol/L PTK787 on cells was strongest, while the continuous increase of PTK787 concentration or prolong of action time did not enhance the inhibitory rate on K562 proliferation. With increasing of drug concentration, the cell proportion in G(1) phase gradually increased, the cell proportion in S phase gradually decreased, the comparison between various groups revealed significant differences (p < 0.05), however the continuous increase of drug concentration from 160 micromol/L did not obviously change the cell proportion in phases of cell cycle. With increasing of drug concentration, the expression of fak mRNA in K562 cells gradually reduced with significant differences between various groups (p < 0.05), but with continuous increase of drug concentration from 160 micromol/L, the effect of PTK787 on the expression of fak mRNA in K562 cells also did not obviously change. It is concluded that the PTK787 shows effect of anti-leukemia cells through inhibiting transformation of the K562 cells from G(1) phase into S phase and decreasing the expression of fak mRNA in cells.
Cell Cycle ; Cell Proliferation ; drug effects ; Focal Adhesion Kinase 1 ; genetics ; Gene Expression Regulation, Leukemic ; Humans ; K562 Cells ; Phthalazines ; pharmacology ; Pyridines ; pharmacology ; RNA, Messenger ; genetics

Objective To investigate the clinical features of these children with congenital hyperinsulinism (CHI) who had no response to diazoxide and provide a theoretical foundation for the formulation of CHI treatment strategy.Methods Eighteen patients with CHI who had no response to diazoxide hospitalized in Beijing Children's Hospital from 2008 to 2012 were chosen as research subjects.Their clinical data were analyzed retrospectively.Results There were 18 patients with persistent hypoglycemia after using diazoxide,which indicated that they had no response to diazoxide.Twelve patients of them were born as macrosomia and their onset age was less than 6 months.Half of the children(9/18 cases) even had hypoglycemia in neonatal period.All the manifestations were conformed to the clinical characteristics of ATP-sensitive potassium channel CHI.Four children who were unresponsive to diazoxide received octreotide treatment,and it was effective on them.Four patients had a near-total pancreatectomy.After a long-term followup study,their blood sugar maintained a normal level,and they did not appear serious postoperative complications.Conclusions Children with CHI who have no response to diazoxide are characterized by coming earlier and higher birth weight.Octreotide is proposed in case of non-response to diazoxide.When medical treatment is not efficient in prevention of hypoglycemia,a subtotal pancreatectomy has to be considered as a last resort.

Objective To study the relationship between cyclic adenosine monophosphate/protein kinase A (cAMP/PKA) pathway and glutamine-stimulated insulin secretion.Methods In the prerequisite of the existence of glucose(0.25 mmol/L),the insulin secretion of βHC9 cells was stimulated with different concentrations of glutamine (0.0,0.5,1.0,2.0,5.0 mmol/L),then culture liquids were extracted and centrifugalized,and the insulin levels in the cell culture liquids and the cAMP levels in βHC9 cells were determined,so as to study the effects of glutamine stimulation on the insulin level in cell culture liquids and cAMP levels in βHC9 cells were assayed.Results In the prerequisite of the glucose existence,with the increasing of the concentrations of glutamine(0.0,0.5,1.0,2.0,5.0 mmol/L),the insulin levels[0.0 ng/(mL · million),19.1 ng/(mL · million),29.1 ng/(mL · million),30.1 ng/(mL · million),33.9 ng/(mL · million)] in cell culture liquids and the cAMP levels (0.0 pmol/million,40.0 pmol/million,51.5 pmol/million,52.5 pmol/million,61.3 pmoL/million) in βHC9 cells increased accordingly.Conclusion Glutamine has amplifying effect on glucose stimulated insulin secretion,such amplifying effect needs the existence of cAMP to be prerequisite.

Antigens, Surface ; analysis ; Antiretroviral Therapy, Highly Active ; CD28 Antigens ; analysis ; CD56 Antigen ; analysis ; HIV Infections ; drug therapy ; immunology ; Humans ; Killer Cells, Natural ; immunology ; Lectins, C-Type ; analysis ; NK Cell Lectin-Like Receptor Subfamily B ; NK Cell Lectin-Like Receptor Subfamily D ; analysis ; Receptors, Immunologic ; analysis ; Receptors, KIR

BACKGROUNDAt the end of 2005, 650,000 people lived with human immunodeficiency virus type-1 (HIV-1) in China, of whom 75 000 were AIDS patients. Many AIDS patients received highly active antiretroviral therapy (HAART) supported by the "China CARES" program but the immune responses of HAART were seldom reported. This study investigated the effect of HAART on the activation and coreceptor expression of T lymphocytes in Chinese HIV/AIDS patients and evaluated its effect on immune reconstitution.

METHODSSeventeen HIV/AIDS patients were enrolled and three-color-flow cytometry was used to detect the activation of HLA-DR CD38 and the coreceptor CCR5, CXCR4 expression on T lymphocytes in whole blood samples taken from the patients before and after 3- or 6-month HAART.

RESULTSThe activation percents of CD4(+), CD8(+) T lymphocytes were significantly higher before therapy than the normal controls (HLA-DR/CD4: 40.47 +/- 18.85 vs 11.54 +/- 4.10; CD38/CD4: 81.34 +/- 10.86 vs 53.34 +/- 11.44; HLA-DR/CD8: 63.94 +/- 12.71 vs 25.67 +/- 9.18; CD38/CD8: 86.56 +/- 11.41 vs 58.84 +/- 6.16, all P < 0.01). After 6-month combined antiretroviral treatment, the activation of T lymphocytes in HIV/AIDS patients was significantly decreased (HLA-DR/CD4: 28.31 +/- 13.48; CD38/CD4: 69.88 +/- 12.64; HLA-DR/CD8: 46.56 +/- 18.64; CD38/CD8: 70.17 +/- 14.54, all P < 0.01 compared with the pre-treatment values). Before the treatment, CCR5 expression on CD8(+) T lymphocytes was up-regulated while CXCR4 expression on CD8(+) T lymphocytes downregulated in HIV/AIDS patients compared with the normal controls (CD8/CCR5: 70.91 +/- 10.03 vs 52.70 +/- 7.68; CD8/CXCR4: 24.14 +/- 11.08 vs 50.05 +/- 11.68, all P < 0.01). After 6-month HAART, CCR5 expression on CD8(+) T lymphocytes significantly decreased (56.35 +/- 12.96, P < 0.01), while CXCR4 expression on CD8(+) T lymphocytes increased (36.95 +/- 9.96, P < 0.05) compared with the pre-treatment and the normal controls. A significant statistical relationship was observed between the expression of activation markers, CCR5 and the CD4(+) T lymphocyte counts after HAART (P < 0.05).

CONCLUSIONSReduced activation of T lymphocytes and a normalization of coreceptor expression were observed in Chinese HIV/AIDS patients after HAART. Immunity can be restored in HIV/AIDS patients receiving HAART.

Acquired Immunodeficiency Syndrome ; drug therapy ; immunology ; Adult ; Antiretroviral Therapy, Highly Active ; China ; Female ; HIV Infections ; drug therapy ; immunology ; Humans ; Lymphocyte Activation ; drug effects ; physiology ; Male ; Middle Aged ; Receptors, Chemokine ; analysis ; drug effects ; T-Lymphocytes ; immunology

OBJECTIVETo explore the effect and mechanism on HBV replication in C gene truncated mutant.

METHODSProtein expression of C gene truncated vector and wild C gene vector were assay by SDS-PAGE Western blot. Constructed C gene truncated expression vector was cotransfected with wild HBV genome; virus load was detected by PCR in the culture medium and the cell. The formation of core particle was assay by Native western blot.

RESULTSThe recombinant vectors can efficiently express. Virus load of the cotransfected group by pcDNA3-deltaC and adwR9 was lower than that of control group in the culture medium and the cell. Protein band of the co-expressed group by pcDNA3-deltaC and pcDNA3-C showed slightly weaker than that of the co-expressed group by pcDNA3 and pcDNA3-C.

CONCLUSIONC gene truncated mutant could interfere with the formation of core particle and reduce of HBV replication

Cell Line ; Genetic Therapy ; Hepatitis B ; therapy ; Humans ; Mutation ; Transfection ; Viral Core Proteins ; genetics ; Virus Replication

OBJECTIVETo study the action mechanism of p,p'-DDE and/or beta-BHC on JNK and MAPK signal transduction pathways in rat Sertoli cells in vitro.

METHODSWe cultured the Sertoli cells isolated from rat testicular tissues for 2 days in vitro, divided them into a control group incubated with DMSO and 3 case groups exposed to p,p'-DDE and / or beta-BHC at the final concentration of 10, 30, 50 micromol/L for 24 hours, and then detected the expression levels of JNK and c-jun mRNA by two-step RT-PCR.

RESULTSTwenty-four hours after p,p'-DDE treatment, the grayscale values of JNK mRNA were 0.068 +/- 0.001, 0.164 +/- 0.002, 0.207 +/- 0.006 and 0.499 +/- 0.017, and those of c-jun mRNA were 0.122 +/- 0.002, 0.157 +/- 0.006, 0.218 +/- 0.007 and 0.289 +/- 0.004 respectively in the DMSO control and the 10, 30 and 50 micromol/L groups. The expressions of JNK and c-jun mRNA were elevated with increased concentration of p,p'-DDE, with significant differences between the control and the case groups (P < 0.05), and they were also significantly upregulated in the beta-BHC and p,p'-DDE + beta-BHC groups in a dose-dependent manner. The grayscale values of JNK mRNA in the p,p'-DDE, beta-BHC and p,p'-DDE + beta-BHC groups at the concentration of 10 micromol/L were 0.164 +/- 0.002, 0.149 +/- 0.003 and 0.178 +/- 0.004, and those of c-jun mRNA were 0.157 +/- 0.006, 0.131 +/- 0.004 and 0.172 +/- 0.002, respectively, both significantly higher in the combination group than in the former two (P < 0.05). And the same was the case with the 30 and 50 micromol/L concentrations.

CONCLUSIONBoth p,p'-DDE and beta-BHC can enhance the expressions of JNK and c-jun in Sertoli cells, and their combination can produce even more obvious effect.

Animals ; Cells, Cultured ; Dichlorodiphenyl Dichloroethylene ; chemistry ; toxicity ; Dose-Response Relationship, Drug ; Gene Expression ; drug effects ; Lindane ; chemistry ; toxicity ; MAP Kinase Signaling System ; drug effects ; Male ; Proto-Oncogene Proteins c-jun ; genetics ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction ; Sertoli Cells ; cytology ; drug effects ; metabolism

OBJECTIVETo collect background information on drug resistance mutations in treatment-naïve HIV-1 infected individuals in Liaoning Province.

METHODSSamples from 91 antiretroviral therapy-naïve patients were collected. The entire protease gene and 1-290 amino acids of the reverse transcriptase gene were amplified by nested PCR from provirus DNA and sequenced. The results were analyzed with HIVdb-Drug Resistance Algorithm, and genotypic resistance mutations were determined to particular anti-HIV drugs.

RESULTSTotally 91 sequences were obtained, 3 of which displayed M46I mutations in the protease gene. Minor resistance mutation rate to protease inhibitors was 100%, including types of L63P (60.4%), V77I (60.4%), M36I/V (31.9%), A71V/T (22.0%), L10I (8.8%), and K20R (6.6%). Only one sequence carried reverse transcriptase related resistance mutations M184I.

CONCLUSIONSAbout 4.4% of HIV-1 infected individuals in Liaoning Province carried strains with drug resistance mutations. Most treatment-naïve HIV-1 infected individuals in Liaoning Province were sensitive to the currently available antiviral medicines, but antiviral treatment must be in accordance with the strict procedures to keep better adherence and avoid the prevalence of drug-resistant strains.

Adult ; China ; epidemiology ; Drug Resistance, Viral ; genetics ; Female ; HIV Infections ; drug therapy ; epidemiology ; HIV Protease ; genetics ; HIV Reverse Transcriptase ; genetics ; HIV-1 ; genetics ; Humans ; Male ; Molecular Epidemiology ; Mutation ; Sequence Analysis, DNA