Objective: To investigate the chemical constituents of Gracilaria lemaneiformis. Methods: Compounds were isolated by normal phase silica gel and Sephadex LH-20 gel column chromatography. Their structures were elucidated by analyses of spectroscopic data. Results: Nine compounds were isolated from the ethanol extract of G. lemaneiformis, their structures were elucidated as palmitic acid (1), 1, 3-dipalmitin (2), 1-monopalmitin (3), uracil (4), adenosine (5), 2α-oxa-2-oxo-5α-hydroxy-3,4-dinorcholestane (6), 1-O-β-D-galactosyl-(6→1)-α-D-galactosyl-2, 3-O-dihexadecanoyl- glycerol (7), 1-O-α-D-glucose-2, 3-O-dihexadecanoyl-glycerol (8), and β-sitosterol (9). Conclusion: All of these compounds are obtained from this plant for the first time.

This study was aimed to propagate and identify the prdm1 gene-knockout mice, so as to lay the foundation for studying Blimp-1 protein. Two kinds of transgenic homozygous mice with B6.prdm1(flox/flox) and B6.Lck-Cre were feed and propagated; after successful propagating, the first passage mice were obtained; after the first passage mice were copulated once again, the genotypes were obtained as follows: B6. prdm1(wild/wild). Lck-Cre, B6. prdm1(wild/wild), B6.prdm1(flox/flox). Lck-Cre, B6.prdm1(flox/wild). Lck-Cre, B6.prdm1(flox/flox), B6. prdm1(flox/wild). The genomic DNA of second passage mice was extracted, the Cre and loxp gene fragments were amplified by PCR, then the size of Cre and loxp genomic DNA were detected by agarose gel electrophoresis. The mice with B6.prdm1(flow/flox). Lek-Cre were used as conditionally prdm1-knockout mice, B6.prdm1(flox/wild). Lck-Cre mice, B6.prdm1(flox/flox) and B6 mice were used as controls. The spleen T lymphocytes and B lymphocytes were sorted by using magnetic beads, the blimp-1 target protein was identified by Western blot. The results showed that the two transgenic homozygous mice had the ability to reproduce, and the separation ratio of second passage mice generated from propagation of their offspring cach other meet Mendelian laws, and the prdm1 gene-knockout mice also could successfully obtained. It is concluded that the application of Cre-loxp system may successfully obtain plentiful prdm1 gene-knockout mice.
Animals ; Genotype ; Mice ; Mice, Inbred C57BL ; genetics ; Mice, Knockout ; genetics ; Reproduction ; Transcription Factors ; genetics

OBJECTIVEThis article reports a population genetic study on six short tandem repeat(STR) loci, D7S820, D19S253, D12S391, D5S818, D16S539 and D8S1179, in a sample of unrelated Chinese Han individuals(n=122-173) living in Hebei province.

METHODSDNA extraction from blood samples (200 in number) and multiplex amplification of the above six loci were carried out. Using denaturing polyacrylamide gel electrophoresis and silver stain, the authors investigated the distribution of allele frequencies of the six loci in Han population in Hebei province.

RESULTSThe STR polymorphisms at all of the six loci were observed in Chinese Han population in Hebei province. The observed heterozygosities of D7S820, D19S253, D12S391, D5S818, D16S539 and D8S1179 were 0.828, 0.757, 0.769, 0.837, 0.785 and 0.852, respectively. The measured values of the power of discrimination (PD) were 0.914, 0.919, 0.940, 0.909, 0.917, 0.944; of the mean exclusion chance(MEC) 0.618, 0.740, 0.801, 0.557, 0.655, 0.696 and of the polymorphism information content (PIC) in Chinese 0.771, 0.760, 0.762, 0.708, 0.776 and 0.794, respectively.

CONCLUSIONThe genotype distributions of the six STR were in accordance with Hardy-Weinberg equilibrium. The numerical values of the PD and MEC are relatively high in Hebei province, and thus can be of significant application in population genetics and forensic medicine.

Asian Continental Ancestry Group ; genetics ; China ; ethnology ; DNA ; analysis ; Female ; Gene Frequency ; Genetics, Population ; Humans ; Male ; Microsatellite Repeats ; genetics ; Polymorphism, Genetic ; Population Groups ; Tandem Repeat Sequences ; genetics ; physiology

OBJECTIVETo investigate the protective effect and mechanism of Yimucao (Herba leonuri) injection against experimental cerebral ischemia.

METHODSMouse models of cerebral ischemia induced by bilateral carotid artery occlusion or potassium cyanide and rat models of middle cerebral artery occlusion (MCAO)-induced cerebral ischemia/reperfusion injury were established to evaluate the protective effect of Yimucao injection by measuring the changes in cerebral malondialdehyde (MDA) content and the activities of superoxide dismutase (SOD) and lactate dehydrogenase (LDH) after the injections.

RESULTSYimucao injection significantly lowered the cerebral index of mice with cerebral ischemia, prolonged the survival time of mice poisoned with potassium cyanide, resulting also in significantly decreased MDA content and increased activities of SOD and LDH in the brain tissue of rats after a 10-min cerebral ischemia followed by 30 min of reperfusion.

CONCLUSIONYimucao injection provides protective effect against experimental cerebral ischemia.

Animals ; Brain Ischemia ; drug therapy ; enzymology ; metabolism ; Disease Models, Animal ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Humans ; L-Lactate Dehydrogenase ; metabolism ; Male ; Malondialdehyde ; metabolism ; Mice ; Protective Agents ; administration & dosage ; Rats ; Rats, Sprague-Dawley ; Superoxide Dismutase ; metabolism

OBJECTIVETo study the curative effects of pirfenidone (PF) on pulmonary fibrosis induced by paraquat (PQ) in mice and to provide the theoretical basis for clinical treatment.

METHODSNinety adult healthy male ICR mice were randomly divided into six groups: control group, PQ group, 2 mg/kg Dexamethasone group, 25 mg/kg PF group, 50 mg/kg PF group and 100 mg/kg PF group, there were 15 mice in each group. The corresponding volume of normal saline was given to the each mouse in control group according to the weight, after 2 h 0.1% CMC was given to the each mouse of control group one time by intragastric administration, then the CMC was administrated at regular time until sacrifice. All mice for other 5 groups were exposed to 100 mg/kg PQ by intragastric administration. At 2 h after exposure to PQ, 0.02 ml/10 g dexamethasone and 25, 50, 100 mg/kg PF were given to mice for dexamethasone group and for 3 PF groups by intragastric administration each day for 49 days, respectively. The lung coefficient was calculated and pathological changes of lung tissue were observed by HE staining for each mouse. The hydroxyproline (HYP) level in lung tissue was measured for each mouse. The mRNA level of and the protein level of TGF-β(1) in lung tissue for each mouse were determined, and the protein level of TGF-β(1) in the bronchus-alveolus lavage fluid (BALF) of each mouse was detected.

RESULTSThe survival rates on the 3rd day in PQ group, 3 PF groups and dexamethasone group were 53.33%, 46.67%, 73.33%, 86.67% and 80%, respectively. The survival rates on the 3rd day in dexamethasone group, 50 mg/kg and 100 mg/kg PF groups were significantly higher than those of PQ group and 25 mg/kg PF group (P < 0.05). The lung coefficients of 3 PF groups were significantly lower than that of the PQ group (P < 0.05). The lung tissue HYP levels of dexamethasone group and 3 PF groups were 50.95 ± 11.65, 44.52 ± 9.48, 43.27 ± 6.01 and 40.82 ± 5.90 mg/g respectively, which were significantly lower than that (74.27 ± 3.68) of PQ group (P < 0.01). The TGF-β(1) protein levels of BALF in dexamethasone group, 50 and 100 mg/kg PF groups were 22.03 ± 7.27, 27.75 ± 5.84 and 21.31 ± 6.82 ng/ml respectively, which were significantly lower than that (52.52 ± 15.51) ng/ml of PQ group (P < 0.01) The expression level of TGF-β(1) mRNA in 100 mg/kg PF group decreased significantly, as compared with PQ group (P < 0.01).

CONCLUSIONPF could reduce the collagen deposition and pulmonary fibrosis induced by PQ in mice lungs.

Animals ; Disease Models, Animal ; Lung ; metabolism ; pathology ; Male ; Mice ; Mice, Inbred ICR ; Paraquat ; poisoning ; Pulmonary Fibrosis ; chemically induced ; drug therapy ; pathology ; Pyridones ; therapeutic use ; Transforming Growth Factor beta ; metabolism

OBJECTIVETo evaluate the status of lymph node metastasis (LNM) and reasonable procedure in early gastric cancer (EGC).

METHODSTwo hundred and ninety-two patients with histologically proven mucosal or submucosal gastric cancer who underwent gastrectomy/lymphadenectomy were included in this study. The numbers of total dissected lymph node were all above 15 in all patients. The clinical characteristics, pathologic features, and LNM were assessed by univariate and multivariate analysis.

RESULTSLNM were observed in 43 of 292 cases (14.7%), and 6.4% in mucosal lesions and 22.4% in submucosal lesions. The LNM was identified in 12.7% at the first level, 7.2% at the second level and 0.34% at the third level. The LNM frequency was found in the 6, 3, 4, 1, 5 lymph node (from high to low) consequently at the first level and the 7, 8a lymph node at the second level. The EGC with nodal involvement and the tumors < 2.0 cm in diameter were all depressed type. The diameter of elevated type with LNM was no less than 3.0 cm in this series. The depth of invasion and lymphatic vessel involvement were independent influencing factors in LNM on multivariate analysis (P<0.05).

CONCLUSIONSLess extensive surgery might be considered for the elevated type EGC, and tumors 3.0 cm in diameter. With respect to the depressed type EGC/tumors > 3.0 cm in diameter, gastrectomy with D2 lymphadenectomy is proper.

Adult ; Aged ; Aged, 80 and over ; Female ; Gastrectomy ; methods ; Humans ; Lymph Node Excision ; methods ; Lymph Nodes ; pathology ; surgery ; Lymphatic Metastasis ; Male ; Middle Aged ; Multivariate Analysis ; Retrospective Studies ; Stomach Neoplasms ; pathology ; surgery

OBJECTIVETo evaluate the effects of antiviral agents on intrahepatic HBV covalently closed circular DNA (cccDNA) in HBeAg-positive chronic hepatitis B patients.

METHODSSeventy-one HBeAg positive chronic hepatitis B patients were enrolled in this study. Lamivudine was administered to 35 patients for 48 weeks, sequential therapy with lamivudine-IFN alpha-2b to 24 of the 71 patients for 48 weeks, and interferon alpha (IFN alpha-2b) was administered to 12 for 24 weeks. All subjects were followed-up for 24 weeks. Serum HBV DNA, intrahepatic HBV DNA and cccDNA were measured quantitatively by PCR. HBV genotypes were analyzed by PCR-RFLP.

RESULTSForty-eight weeks of sequential lamivudine-IFN alpha-therapy and lamivudine monotherapy and 24 weeks of IFN alpha monotherapy reduced the intrahepatic HBV DNA to (4.7+/-1.1) log10, (4.6+/-1.5) log10 and (5.6+/-1.5) log10, and cccDNA to (3.4+/-1.3) log10, (3.8+/-1.1) log10 and (5.0+/-1.5) log10, significantly lower than therapy (P < 0.05). Seventeen of the 71 patients developed HBeAg seroconversion, and the reduction of cccDNA in the HBeAg seroconverted patients was significantly more than that of the HBeAg positive patients (P < 0.05). After 24 weeks of antiviral therapy withdrawal, 18 patients achieved sustained virological response, and the baseline intrahepatic cccDNA in the patients with sustained virological response was significantly lower than that of patients with virological rebound (P < 0.05). The change in intrahepatic cccDNA correlated positively with the reduction in intrahepatic HBV DNA (P < 0.05). The cccDNA levels correlated with the serum HBeAg titers at the end of the treatment (P < 0.01). Of the total 71 cases, HBV genotype C accounted for 85.9% (n = 61), and genotype B for 14.1% (n = 10). There was no significant difference in the changes of intrahepatic HBV DNA and cccDNA levels between HBV genotypes C and B (P >0.05).

CONCLUSIONSBoth 48 weeks of sequential lamivudine-IFN alpha and lamivudine monotherapy strongly reduced intrahepatic HBV DNA and cccDNA more than 24 weeks of IFN alpha monotherapy. Low baseline intrahepatic cccDNA levels might predict a good long-term efficacy of antiviral treatment. The reduction of intrahepatic cccDNA correlated positively with the changes of intrahepatic HBV DNA, and intrahepatic cccDNA levels correlated with serum HBeAg titers. HBV genotypes had no obvious influence on intrahepatic HBV DNA load or cccDNA load.

Adult ; Antiviral Agents ; pharmacology ; therapeutic use ; DNA, Circular ; DNA, Viral ; Female ; Hepatitis B e Antigens ; blood ; Hepatitis B virus ; drug effects ; genetics ; Hepatitis B, Chronic ; drug therapy ; virology ; Humans ; Interferon-alpha ; pharmacology ; therapeutic use ; Lamivudine ; pharmacology ; therapeutic use ; Male ; Middle Aged ; Recombinant Proteins ; Young Adult

AIMTo investigate the antioxidant mechanisms of propylene glycol mannate sulfate (PGMS) in hyperlipidemic rats.

METHODSMale Wistar rats were given high lipid emulsion diet to establish hyperlipidemic model. PGMS was given every day at different doses (37.8 and 75.6 mg.kg-1, ig) to hyperlipidemic rats for three weeks. In addition, diethyldithiocarbamate (DDC) was given 200 mg.kg-1.3 d-1 (i.p.) to inhibit SOD activity. Then, the MDA content was examined using TBA method to show the oxidation level, and the activities of SOD, GSH-Px and CAT were examined following the kit protocols to indicate the capability of eliminating OFR. RT-PCR was applied to study the expression of Cu, Zn-SOD mRNA in rat liver.

RESULTSThe MDA content of PGMS treatment groups decreased markedly compared with hyperlipidemic group, and the activities of SOD, GSH-Px and CAT increased distinctly. Cu, Zn-SOD mRNA expression was significantly increased by PGMS treatment. Furthermore, the application of DDC(the SOD inhibitor) reduced total SOD activity and Cu, Zn-SOD mRNA expression induced by PGMS, and the content of MDA increased correspondingly.

CONCLUSIONPGMS can induce the activities of antioxidant enzymes and the mRNA expression of Cu, Zn-SOD, which contribute to the elimination of oxygen free radical. This may explain the molecular mechanism of antioxidant effects of PGMS.

Animals ; Antioxidants ; pharmacology ; Catalase ; metabolism ; Free Radical Scavengers ; pharmacology ; Hyperlipidemias ; enzymology ; metabolism ; Liver ; enzymology ; metabolism ; Male ; Malondialdehyde ; metabolism ; Propylene Glycols ; pharmacology ; RNA, Messenger ; genetics ; Rats ; Rats, Wistar ; Superoxide Dismutase ; biosynthesis ; genetics ; metabolism

AIM:To explore the effect of curcumin(Cur)and curcuminoids(Y20 and 6B)on the expression of secretory leukocyte protease inhibitor(SLPI), tumor necrosis factor-α(TNF-α)and interleukin-1β(IL-1β)induced by Streptococcus pneumoniae(SP)and the possible mechanism.METHODS:BEAS-2B cells incubated with SP were set up as an inflammation model of pneumonia.The mRNA levels of SLPI at 1 h,3 h,6 h and 9 h,and the mRNA expression of TNF-αand IL-1βat 3 h,6 h and 9 h in control group,SP infection group,Cur treatment group,Y20 treatment group and 6B treatment group were measured by qPCR.The protein levels of TNF-αand IL-1βin the culture supernatant were measured by ELISA.The protein levels of Toll-like receptor 2(TLR2)and phosphorylated nuclear factor-κB(p-NF-κB) p65 at 3 h,6 h and 9 h were determined by Western blot.RESULTS:The mRNA level of SLPI was increased in Cur, Y20 and 6B treatment groups compared with SP group(P<0.05).The protein levels of TLR2 and p-NF-κB p65 were sig-nificantly increased after SP stimulation.After treatment with Cur,Y20 and 6B,the protein levels of TLR2 and p-NF-κB p65 were significantly decreased(P<0.05).The levels of TNF-αand IL-1βwere significantly increased after SP stimula-tion.Cur,Y20 and 6B significantly decreased the levels of TNF-αand IL-1βin the supernatant(P<0.05).CONCLU-SION: Cur, Y20 and 6B increase SLPI expression, reduce the expression of inflammatory cytokines TNF-αand IL-1β. The possible mechanism might be associated with inhibiting TLR 2 expression and down-regulating the transcriptional activity of NF-κB.

Objective To investigate clinical effect and indications of neuroendoscopic surgery for treatment of intracranial diseases. Methods A total of 386 cases treated with neuroendoscopic surgeries were analyzedd retrospectively in regard to the surgical approaches, treatment outcomes, advantages and disadvantages of neuroendoscopy. Results In the 386 cases, clinical cure was achieved in 340 cases and symptomatic improvement in 40 cases; 5 cases failed to respond to the treatment, and death occurred in 1 case during the follow-up for 6-12 months. Postoperative radiography demonstrated intracranial space-occupying lesions in 285 cases, for which total resection was achieved in 259 cases and subtotal resection in 26 cases with neuroendoscopic surgery, and relapse occurred in 7 cases. Hydrocephalus occurred in 101 cases. Cerebral ventricle recovery was found in 82 cases, and in 18 cases the ventricular expansion was obviously reduced; ventricular expansion remained unchanged in 1 case after the treatment. Conclusions With careful evaluation of the surgical indications, neuroendoscopic surgeries can achieve satisfactory effects for treatment of intracranial diseases.