Objective To investigate the age-associated changes of ultrastructure,mRNA and protein expressions of H+-K+-ATPase in elderly gastric parietal cell. Methods Fifty patients with relative normal stomach without gastroduodenal diseases were enrolled,including younger group (aged 20-59 years,n=19) and elderly group (aged≥60 years,n=31).Furthermore,the elderly group was divided into 3 subgroups:60-69 years old (n =11 ),70-79 years old (n=10 ),above 80 years old (n =10).The ultrastructure of gastric parietal cell was observed under electron microscope.The expression of H+-K+-ATPase α subunit mRNA and H+-K+-ATPase β subunit protein were assessed by quantitative real-time PCR and Western-blot,respectively.The ageing-associated changes of all these data were respectively compared. Results No significant difference was showed in the morphology of gastric parietal cell and acid-secretion-associated organelles among all the groups.The average ratio Am to Ac (Am means the area of mitochondria,Ac means the area of cytoplasm) of gastric parietal cell and the average At to Ac ratio (At means the area of secretory canaliculi and tubulovesicular system )between younger group and elderly group had no significant difference[(48.4±7.5) ％ vs.(50.6±7.6) ％,t=-0.775,P=0.444; (13.8±4.1) ％ vs.(12.2±4.7) ％,t=0.984,P=0.332].Meanwhile,there were no distinctions in the expression of H+-K+ -ATPase α subunit mRNA and H+-K+-ATPase protein among all elderly subgroups(F=1.522,2.32,P=0.24,0.114).However,the mRNA expression of H+-K+-ATPase a subunit was higher in the elderly group than in the younger group(t=-3.682,P=0.001).Furthermore,the expression of H+ -K+ -ATPase protein in the elderly group was increased as compared with younger group(t=-3.389,P=0.004). Conclusions Acidsecretion-associated organelles of human gastric parietal cell have no degeneration and the expression of H + -K+-ATPase is in trend of increase with aging,indicating that healthy elderly people have the basis of ultrastructure and molecular biology to maintain well function of acid secretion.

According to the law of circulation of meridians and the locations of acupoints, the opposite acupoints were proposed. It facilitates comprehension of the routes of meridians and the locations of acupoints. Application of opposite needling or penetrative needling, it is easy to practice and the effectiveness is significant. Promoting this concept into acupuncture training, can expand out from acupoint to meridian, from one meridian to other meridians, it will generate good rewards.
Acupuncture ; education ; methods ; Acupuncture Points ; Meridians ; Teaching ; methods

AIMTo prepare buspirone hydrochloride buccal adhesive tablet and investigate factors that influence drug release behavior and the drug release mechanism.

METHODSBuspirone hydrochloride buccal adhesive tablet was prepared with double layers structure composed of drug core and adhesive layer. The materials of the drug core were carbopol 974 and lactose, the adhesive layers were carbopol 974 and HPMC K4M. The influence of drug release factors such as adhesive layer component, adhesive layer weight and adhesive tablet hardness was investigated. The relationship between adhesive layer weight and drug release mechanism in vitro was studied.

RESULTSThe results showed that the weight of the adhesive layer and the hardness of adhesive tablet showed significant effects on drug release, but the adhesive layer component showed no significant effect. The optimum prescription of buspirone hydrochloride buccal adhesive tablet was carbopol: HPMC = 1:1, adhesive layer weight = 50%, and adhesive tablet hardness = 4 kg. The study of drug release mechanism from adhesive tablet showed that it was double directions when adhesive layer weight was 20%, and single direction first then double directions when 33.33%, and single direction all along when 50%.

CONCLUSIONThe speed and direction of drug release from adhesive tablet can be controlled by regulating adhesive layer weight.

Acrylic Resins ; Adhesiveness ; Administration, Buccal ; Anti-Anxiety Agents ; administration & dosage ; Buspirone ; administration & dosage ; Delayed-Action Preparations ; Drug Carriers ; Hardness ; Lactose ; analogs & derivatives ; chemistry ; Methylcellulose ; analogs & derivatives ; chemistry ; Oxazines ; Polyvinyls ; chemistry ; Tablets ; Technology, Pharmaceutical

Objective To identify mutations site and clinical characteristics of a familial hypercholesterolemia(FH) proband diagnosed clinically through DNA sequencing and family analysis in the proband and his family members of 3 generations.Methods Blood samples and clinical data of the kindred of total 29 from 3 generations members were collected.Proband had a physical examination electrocar-diogrom and vascular ultrasound.The proband and his family members took routine clinical exams,and genomic DNA was isolated.The promoter region and the 18 exons of low density liporotein receptor(LDLR) gene were screened by Touch down polymerase chain reaction -single strand conformation polymorphism(PCR-SSCP) and DNA sequencing.The result of sequencing were matched gene sequence published in the BLAST database.Results 1.Increased intima-media thickness and plaque were detected in the common carotid artery,right subclavian artery of the proband.Aortic valve regurgitation was found by echocardiography.2.No mutation R3500Q of ApoB100 was observed.3.Two heterozygous mutations in exon 10 and 13 of LDLR gene (W462X and A606T) were identified.The proband and 5 members of paternal relatives showed W462X heterozygosis mutation in exon 10 of LDLR gene which introduced the change from tryptophone to a new stop codon.The proband's mother and grandmother harboured A606T heterozygous mutation in exon 13 of LDLR gene due to a single base pair substitution of G for A in the codon for residue 1 879.Conclusions Disease causing mutations of proband are W462X and A606T compound heterozygosis mutation in exon 10 and 13 of LDLR gene inherited from mother and father.Proband shows homozyous phenotype though the genotype analysis indicates heterozygous mutations.

OBJECTIVETo probe into the bidirectional regulation of acupuncture in the rat with motility abnormality of the stomach and the nerve mechanism.

METHODSAdopting electrophysiological methods, the extracellular discharge signals of neuronic action in NTS were recorded with glass microelectrodes. The effects of acupuncture at "Zusanli" (ST 36), "Neiguan" (PC 6), "Zhongwan" (CV 12), "Qihai" (CV 6) on discharges of NTS neurons and intra-gastric pressure were observed in the rat at normal physiological state, and increase or reduction of gastric motility.

RESULTSAcupuncture at "Zusanli" (ST 36), "Neiguan" (PC 6) could significantly promote gastric motility of the rat at normal status, increase or reduction of gastric motion, while acupuncture at "Zhongwan" (CV 12), "Qihai" (CV 6) could significantly inhibit gastric motion. And acupuncture at all the above acupoints could activate discharges of NTS neurons.

CONCLUSIONThe bidirectional regulation effect of an acupoint does not certainly indicate that the same point has bidirectional regulation effect on an internal organs at different pathological states, while this may show synthetic action of different points which is a good regulation towards normal states and homeostasis.

Acupuncture Points ; Acupuncture Therapy ; Animals ; Gastrointestinal Motility ; Male ; Rats ; Rats, Sprague-Dawley ; Solitary Nucleus ; physiology ; Stomach Diseases ; physiopathology ; therapy

BACKGROUNDTo observe the effects of short-term antibiotic treatment in patients with hepatic failure and spontaneous bacterial peritonitis (SBP).

METHODSIn this prospective study short-term antibiotic treatment was given to 67 cases diagnosed as hepatic failure with spontaneous bacterial peritonitis. Ceftriaxone 2 g, iv drip, q12h for 10 d or ofloxacin 0.2 g, iv drip, q12h for 10 d was given to the patients at random and the efficacy was evaluated on the basis of clinical symptoms, medical examination and ascites after 3, 7, 10 days of therapy.

RESULTSSeven cases (10.44%) were cured and 57 cases (85%) were improved after 3 days therapy, the total effective rate was 95.52%, but in 3 cases the therapy had no effect. The results of ascites bacterial culture and drug susceptibility test showed that one case had drug resistance to ceftriaxone and two cases had drug resistance to ofloxacin, so antibiotics were changed in time. After 7 days therapy, the results showed that 65 cases (97.01%) cured and 2 cases (2.99%) were improved, the total effective rate was 100%. When the therapy lasted for 10 days, all patients were cured. One patient had oral mucous membrane. Candida albicans infection after 3 days therapy; two cases got thrush and one patient got fungal intestinal infection after 7 days therapy; when the therapy lasted for 10 days, 4 cases had thrush and 2 cases had fungal infection of intestines and one patient had pulmonary fungal infection.

CONCLUSIONThe optimal period of antibiotic treatment of hepatic failure with SBP should be from 7 days to 10 days.

Adult ; Anti-Bacterial Agents ; administration & dosage ; Bacterial Infections ; drug therapy ; etiology ; Ceftriaxone ; administration & dosage ; Drug Therapy, Combination ; Female ; Humans ; Liver Failure ; complications ; Male ; Middle Aged ; Ofloxacin ; administration & dosage ; Peritonitis ; drug therapy ; etiology ; Prospective Studies ; Treatment Outcome

Zuotai (gTso thal) is a typical representative of Tibetan medicines containing heavy metals, but there is still lack of modem safety evaluation data so far. In this study, acute toxicity test, sub-acute toxicity test, one-time administration mercury distribution experiment, long-term mercury accumulative toxicity experiment and preliminary study on clinical safety of Compound Dangzuo were conducted in the hope of obtain the medicinal safety data of Zuotai. In the acute toxicity test, half of KM mice given the lethal dose of Zuotai were not died or poisoned, and LD50 was not found. The maximum tolerated dose of Zuotai was 80 g x kg(-1). In the subacute toxicity test, Zuotai could reduce ALT, AST, Crea levels in serums under low dose (13.34 mg x kg(-1) x d(-1)) and medium dose (53.36 mg x kg(-1) x d(-1)), with significant difference under low dose, and increase the levels of ALT, AST, MDA, Crea in serums under high dose (2 000 mg x kg(-1) x d(-1)); besides, the levels of BUN and GSH in serums reduced with the increase in dose of Zuotai, indicating a significant dose-effect relationship. In the one-time administration distribution experiment, the content of mercury in rat kidney, liver and lung increased after the one-time administration with Zuotai, with a significant dose-dependent relationship in kidney. In the long-term mercury accumulative toxicity experiment, KM mice were administered with equivalent doses of Zuotai for 4.5 months and then stopped drug administration for 1.5 months. Since the 2.5th month, they showed significant mercury accumulation in kidney, which gradually reduced after drug withdrawal, without significant change in mercury content in liver, spleen and brain and ALT, AST, TBIL, BUN and Crea in serum. At the 4.5th month after drug administration, KM mice showed slight structural changes in kidney, liver and spleen tissues, and gradually recovered to normal after drug withdrawal. Besides, no significant difference in weight gain was found between the Zuotai group and the control group. According to the findings of the clinical safety study of Dangzuo, after subjects administered Dangzuo under clinical dose for one month, their serum biochemical indicators, blood routine indicators and urine routine indicators showed no significant adverse change. This study proved that traditional Tibetan medicine Zuotai was slightly toxic, with a better safety in clinical combined administration and no adverse effects on bodies under the clinical dose and clinical medication cycle. However, long-term high-dose administration of Zuotai may have a certain effect on kidney.
Adult ; Animals ; Clinical Trials as Topic ; Drugs, Chinese Herbal ; analysis ; pharmacokinetics ; toxicity ; Female ; Humans ; Kidney ; drug effects ; Liver ; drug effects ; Male ; Medicine, Tibetan Traditional ; Mice ; Middle Aged ; Rats ; Rats, Wistar ; Young Adult

OBJECTIVETo study Chinese medicine (CM) signs and symptoms of urethane-induced lung cancer in mice, and observe the effect of Aconiti Lateralis Radix Praeparata and Taraxaci Herba on symptoms in mice and tumor progress.

METHODThe mice were intraperitoneally injected with urethane twice a week for consecutively five weeks to establish a lung cancer model. The changes in their appearance, body temperature and auricle microcirculation were observed in carcinogenic process. CM signs and symptoms of urethane-induced lung cancer in mice were evaluated with energy metabolism, erythrocytic ATP emzymatic activity and hemorrheological index. During the tumor model was induced, Aconiti Lateralis Radix Praeparata and Taraxaci Herba were used to treat the mice and observe their effect on symptoms in mice and tumor progress.

RESULTDuring urethane was used to induce lung cancer, the mice had gradually become chill, lazy, hunched, with reduction in temperature, cyanosis in auricle and tail. Meanwhile, their energy metabolism and erythrocytic ATP enzymatic activity reduced, whereas their whole blood viscosity and erythrocytic aggregate index increased. Taraxaci Herba showed an effect on enhancing above symptoms and signs but had no effect on tumor progress. Aconiti Lateralis Radix Praeparata showed an effect on reducing above symptoms and signs and preventing tumor progress.

CONCLUSIONMice with urethane-induced lung cancer show CM signs and symptoms of congealing cold with blood stasis. The treatment with Aconiti Lateralis Radix Praeparata can alleviate symptoms and signs in mice and prevent tumor progress.

Aconitum ; chemistry ; Animals ; Blood Circulation ; drug effects ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Humans ; Lung Neoplasms ; chemically induced ; drug therapy ; pathology ; physiopathology ; Mice ; Mice, Inbred BALB C ; Neoplastic Processes ; Taraxacum ; chemistry ; Urethane ; adverse effects

BACKGROUNDTo construct human papillomavirus type 18 (HPV18 E6E7) adeno-associated virus (AAV) for studying the role of HPV E6E7 in the development of human cancer.

METHODSHPV18 E6E7 genes were inserted into adeno-associated virus expression vector and then infected 293 cell line. The expression of HPV18 E6E7 genes were confirmed by using RT-PCR/Southern blot assay.

RESULTSThere was HPV18 E6E7 genes in the malignantly transformed cell line. The 293TL cells compared with the parent cells transformed cells grew more rapidly, lost their contact inhibition and formed more and large colonies in soft agar.

CONCLUSIONSHPV18 E6E7 AAV was successfully constructed and could induce malignant transformation. HPV18 E6E7 AAV can be use for studying the immortalization and malignant transformation of human normal epithelial cells.

Cell Line ; Cell Transformation, Neoplastic ; Cell Transformation, Viral ; DNA, Viral ; analysis ; DNA-Binding Proteins ; Dependovirus ; genetics ; Epithelial Cells ; cytology ; virology ; Fetus ; Humans ; Kidney ; cytology ; Oncogene Proteins, Viral ; genetics ; Papillomaviridae ; genetics ; Polymerase Chain Reaction

OBJECTIVETo evaluate the value of PowerPlex21 System (Promega) and study the genetic polymorphism of its 20 short-tandem repeat (STR) loci in southern Chinese Han population.

METHODSWe conducted genotyping experiments using PowerPlex21 System on 20 autosomal STR loci (D3S1358, D1S1656, D6S1043, D13S317, Penta E, D16S539, D18S51, D2S1338, CSF1PO, Penta D, TH01, vWA, D21S11, D7S820, D5S818, TPOX, D8S1179, D12S391, D19S433 and FGA) in 2367 unrelated Chinese Han individuals living in South China. The allele frequencies and parameters commonly used in forensic science were statistically analyzed in these individuals and compared with the reported data of other populations.

RESULTSThe PowerPlex21 System had a power of discrimination (PD) ranging from 0.7839 to 0.9852 and a power of exclusion (PE) ranging from 0.2974 to 0.8099 for the 20 loci. No significant deviation from Hardy-Weinberg expectations was found for all the loci except for D5S818. This southern Chinese Han population had significant differences in the allele frequencies from 8 ethnic groups reported in China, and showed significant differences at 8 to 20 STR foci from 5 foreign populations. The allele frequency at the locus D1S1656 in this southern Chinese Han population differed significantly from those in the 5 foreign populations and from 3 reported Han populations in Beijing, Zhejiang Province and Fujian Province of China. The neighbor-joining phylogenetictree showed clustering of all the Asian populations in one branch, while the northern Italian and Argentina populations clustered in a separate branch. This southern Chinese Han population had the nearest affinity with the Yi ethnic population in Yunnan Province of China.

CONCLUSIONThe 20 STR loci are highly polymorphic in this southern Chinese Han population, suggesting the value of this set of STR loci in forensic personal identification, paternity testing and anthropological study.

Asian Continental Ancestry Group ; genetics ; China ; Cluster Analysis ; Ethnic Groups ; Forensic Sciences ; Gene Frequency ; Genetics, Population ; Genotype ; Humans ; Microsatellite Repeats ; Polymorphism, Genetic