To obtain chemical constituent information of rat plasma after oral administration of Huang-Lian-Jie-Du Decoction (HLJDD), a LC-FT-ICR-MS method has been established, and both positive and negative ions scan modes were include in the analysis. By comparing their retention time, high resolution mass data of HLJDD extracts, blank plasma and dosed plasma, 38 constituents, including 22 prototype compounds and 16 metabolites, were detected in rat plasma after oral administration of HLJDD. In the 22 prototype compounds, 16 constituents were determined unambiguously by comparing with references. In the analysis of metabolites, phase II reactions like glucuronidation and sulfation were the major biotransformation pathways of HLJDD. M11 was observed as the only phase I metabolite in present experiment. The results will be beneficial for the further pharmacokinetics and pharmacological evaluations of HLJDD.
Administration, Oral ; Alkaloids ; blood ; Animals ; Biotransformation ; Chromatography, High Pressure Liquid ; Drug Combinations ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; metabolism ; Flavonoids ; blood ; Iridoids ; blood ; Male ; Plants, Medicinal ; chemistry ; Rats ; Rats, Sprague-Dawley ; Spectrometry, Mass, Electrospray Ionization ; Tandem Mass Spectrometry

Increasing attention has been focused on the antitumor activity of artemisinin derivatives in recent years, for artemisinin had been reported to have cytotoxic effects against HL-60, P388 and MCF-7 tumor cells. We report here the synthesis and evaluation for antitumor activity of a series of artemisinin-ether derivatives bearing tetrahydropyrrole, morpholine, piperidine, substituted piperidines and azoles with various linkers. Sixteen 10-O-substituted dihydroartemisinin derivatives were designed and synthesized, all of which have never been reported in literatures and whose antiproliferative effects on human breast cancer MCF-7, MCF-7/Adr and HL-60 cells were determined by MTT assay or direct cell counting. Each of these artemisinin derivatives possessed better effects than dihydroartemisinin evidently against HL-60 and MCF-7 cells growth, while less potent than doxorubicin. All target compounds exhibited significantly improved potency compared to DHA and doxorubicin on the doxorubicin-resistant MCF-7/Adr cells, so did they in their sensitive counterparts MCF-7 cells. Among them, compounds GF02, GH04 and ZH04 showed strong activity against these three cell lines growth. Further research is undergoing.
Antineoplastic Agents ; chemical synthesis ; chemistry ; Artemisinins ; chemical synthesis ; chemistry ; Breast Neoplasms ; pathology ; Cell Proliferation ; Doxorubicin ; Drug Design ; HL-60 Cells ; drug effects ; Humans ; MCF-7 Cells ; drug effects

OBJECTIVETo consider the relationship to survival rate and quality of life with pararectal lymphadenectomy for lower carcinoma of rectum.

METHODSThe radical operation was performed on 780 cases of progressive cancer located at peritoneal reflection or below it, Among them, 352 cases only cleared in abdominal cavity, 428 cases coupled with extra-peritoneal histopathological type.

RESULTSUrinary function injured, the group cleared in abdominal cavity was 12 cases, accounted for 3.6%; the group coupled with extra-peritoneal clearance was 225 cases, for 52.5% (P < 0.01). Sexual function damaged (only for male), the abdominal cavity group was 23 cases, for 12.6% (23/185); the coupled group was 127 cases, for 53.4% (127/238), (P < 0.01). Local relapse rate, the abdominal cavity group was 15.8% (56/352); the coupled group was 8.6% (41/428), (P < 0.05). 5-year survival rate, the abdominal cavity group was 52.2%; the coupled group was 58.5% (P < 0.05).

CONCLUSIONBy contrast, although abdominal cavity coupled with extraperitoneal lymphadenectomy acted to cut down local relapse and to elevate 5-year survival rate, the postoperative quality of life appeared to be seriously affected.

Female ; Humans ; Lymph Node Excision ; methods ; Male ; Proctocolectomy, Restorative ; Quality of Life ; Rectal Neoplasms ; mortality ; pathology ; surgery ; Retrospective Studies ; Survival Rate

OBJECTIVE:To establish a method for simultaneous determination of tanshinol,caffeic acid,rosmarinic,salviano-lic acid B,salvianolic acid A,tanshinoneⅠ,cryptotanshinone,tanshinone ⅡA and ursolic acid in Compound xueshuantong cap-sules. METHODS:UPLC-MS/MS method was adopted. The determination was performed on ACQUITY UPLC? BEH C18 column with mobile phase consisted of acetonitrile-0.1%formic acid(gradient elution)at the flow rate of 0.2 mL/min. The column tempera-ture was 40 ℃,and the temperature of injector was 10 ℃. Analysis time was 7 min,and sample size was 5 μL. The electrospray ionization source(ESI)was used;ion source temperature was 150℃;capillary voltage was 3.5 kV;cone flow was 50 L/h;desol-vation temperature was 350 ℃;desolvation gas flow was 650 L/h;nebuliser pressure was 7 × 105 Pa;ion monitoring and multiple reaction monitoring (MRM) was performed. RESULTS:The linear ranges of tanshinol,caffeic acid,rosmarinic,salvianolic acid B,salvianolic acid A,tanshinoneⅠ,cryptotanshinone,tanshinone ⅡA and ursolic acid were 10.0-100.0 μg/mL (r=0.9998), 0.1-1.0 μg/mL(r=0.9998),4.0-40.0 μg/mL(r=0.9999),10.0-100.0 μg/mL(r=0.9999),15.0-150.0 μg/mL(r=0.9997), 8.0-80.0 μg/mL(r=0.9998),10.0-100.0 μg/mL(r=0.9997),50.0-500.0 μg/mL(r=0.9997)and 6.0-60.0 μg/mL(r=0.9998), respectively. The limits of quantitation were 40.0,9.6,38.0,88.0,130.0,39.0,4.4,3.2 and 10.0 ng/mL,separately. The limits of detection were 12.0,3.0,11.0,26.0,39.0,12.0,1.3,1.0 and 3.0 ng/mL,respectively. RSDs of precision,stability and repro-ducibility tests were all lower than 3%. The recoveries were 97.34%-103.20%(RSD=2.19%,n=6),97.22%-102.39%(RSD=2.03%,n=6),98.51%-101.70%(RSD=1.32%,n=6),97.86%-102.49%(RSD=2.09%,n=6),96.75%-103.12%(RSD=2.36%,n=6),98.43%-101.65%(RSD=1.25%,n=6), 97.59%-101.50%(RSD=1.50%,n=6), 96.45%-102.88%(RSD=2.58%,n=6),97.02%-103.11%(RSD=2.38%,n=6),separately. CONCLUSIONS:The method is simple and accurate,and can be used for simultaneous determination of 9 components in Compound xueshuantong capsules.

OBJECTIVETo investigate the essentials of operation and the postoperative effect of right hemicolonic carcinoma with anterograde clearance of lymph nodes.

METHODSOne hundred and thirty-five patients with right hemicolonic infiltrated carcinoma, who were eligible for radical excision (D(3)), were divided into 2 groups. Among them, the anterograde clearance of lymph nodes was performed on 56 cases; the retrograde clearance was performed on 79 cases. Both groups showed no significant difference in age, sex, Dukes' staging and pathological type.

RESULTSThe average time of operation: the anterograde group was (180 +/- 40) min; the retrograde group was (180 +/- 20) min. The average amount of bleeding: the anterograde group was (200 +/- 80) ml; the retrograde group was (200 +/- 30) ml. The cleared number of lymph nodes: the anterograde group were 6.3 +/- 4.2, 2.6 +/- 3.1, 1.5 +/- 2.3 in paracolon, middle and radicel of vasorum respectively, the total number was 11.4 +/- 8.6; the retrograde group were 6.4 +/- 2.2, 2.8 +/- 2.1, 1.1 +/- 1.1 respectively, the total number was 10.8 +/- 5.6 (P > 0.05). The postoperative metastasis to liver: the anterograde group was 8 cases (13.9%); the retrograde group was 21 cases (26.6%, P < 0.05). The 5-year survival rate: the anterograde group was 72.8% (41/56); the retrograde group was 65.5% (52/79) (P < 0.05).

CONCLUSIONSThe operative technique of the excision was little difficulty and complexity, and it could fit well with the requirement of non touch isolation, and act to cut down the postoperative metastasis to liver and to elevate 5-year survival rate.

Adult ; Colonic Neoplasms ; mortality ; surgery ; Female ; Follow-Up Studies ; Humans ; Lymph Node Excision ; methods ; Male ; Middle Aged ; Retrospective Studies ; Survival Rate ; Treatment Outcome

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

OBJECTIVETo investigate the feasibility of using modified peroneal tendofascial compound flap with partial tendon of peroneal long muscle in lower leg refractory defect with Achilles tendon rupture.

METHODSBased on the application experience of peroneal tendofascial compound flap for lower leg refractory defects, partial tendon of peroneal long muscle was included in the flap to repair the Achilles tendon rupture.

RESULTS6 cases were treated. The maximum size of modified compound flap reached 5 cm x 5 cm. The maximal included tendon in the flap was 5 cm in length. All the flaps survived completely. Partial epidermis necrosis happened in one case between the skin graft at the donor site and the edge of the flap pedicle. The patients were followed up for 5 months to 2 years with satisfactory cosmetic and functional results.

CONCLUSIONSThe modified compound flap has reliable blood supply and can be easily transposed, leaving less morbidity at the donor site. It is very suitable for lower leg refractory defect with the Achilles tendon rupture.

Achilles Tendon ; injuries ; Adult ; Aged ; Feasibility Studies ; Female ; Follow-Up Studies ; Humans ; Leg Injuries ; surgery ; Male ; Middle Aged ; Surgical Flaps ; Tendon Injuries ; surgery ; Treatment Outcome

The purpose of this study is to investigate the Sal I, Nru I and Mse I restriction fragment length polymorphisms (RFLPs) of factor IX gene in Chinese Han people. The frequencies of FIX-192 and FIX-793 for A and G, and FIX-698 for T and C were analyzed by polymerase chain reaction (PCR) in unrelated normal Chinese Han people. A sample of 214, 210 and 206 unrelated X chromosomes were analyzed for FIX-192 and FIX-793 and FIX-698, respectively. The results showed that the frequencies for FIX-192 were 0.878 for A and 0.122 for G, with a heterozygosity rate of 0.213, and the frequencies for FIX-793 were 0.552 for A and 0.448 for G, with a heterozygosity rate of 0.494, the frequencies for FIX-698 were 0.311 for T and 0.689 for C, with a heterozygosity rate of 0.429. It was concluded that the SalIand NruI and MseI RFLPs of FIX gene may be useful markers for carrier detection and prenatal diagnosis in Chinese families with hemophilia B patients.
China ; DNA ; genetics ; metabolism ; Deoxyribonucleases, Type II Site-Specific ; metabolism ; Factor IX ; genetics ; Female ; Gene Frequency ; Heterozygote ; Humans ; Male ; Polymorphism, Restriction Fragment Length

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 investigate the effects of saikosaponin a (SSa) on Glu-activated hippocampal astrocytes of rats.

METHODSNeonatal rat (1-3 days) hippocampal astrocytes were obtained and divided into control group, L-Glu activation group and SSa groups with SSa treatment at 5, 2.5, and 1.25 mg/L. The cell proliferation, cell cycle changes, and expression of glial fibrillary acidic protein (GFAP) after the treatments were assessed with MTT assay, flow cytometry and Western blotting, respectively.

RESULTSIn comparison with Glu-activation group, SSa treatment resulted in significant inhibition of the cell proliferation, cell division and GFAP expression in the Glu-activated astrocytes (P < 0.05). SSa at 2.5 mg/L showed the strongest inhibitory effects against astrocyte activation and maintained nearly normal level of astrocyte activation in comparison with the control group (P > 0.05).

CONCLUSIONSGlu-induced activation of rat hippocampal astrocytes can be inhibited by SSa, whose antiepileptic effects is probably mediated by inhibition of hippocampal astrocyte activation.

Animals ; Animals, Newborn ; Astrocytes ; cytology ; Cell Proliferation ; drug effects ; Cells, Cultured ; Female ; Glial Fibrillary Acidic Protein ; biosynthesis ; Glutamic Acid ; pharmacology ; Hippocampus ; cytology ; Male ; Oleanolic Acid ; analogs & derivatives ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Saponins ; pharmacology