OBJECTIVETo describe a HPLC method for assessing betaine in Fufang Guilu granule.

METHODThe content of betainephenaxcyl bromide in Fufang Guilu granule was determined by HPLC. The analytical column was a shim-pack CLC-ODS (6.0 mm x 150 mm) filling a 5 microm stationary phase; The mobile phase consisted of acetonitrile-water(35:65) with 0.1 mol x L(-1) NaClO4; The flow-rate was 1 mL x min (-1); The detector was set at 254 nm.

RESULTThe calibration curve was linear over the range of 0.09-0.585 microg (r = 0.9997). The average recovery of the method was 98.4%, RSD 2.5% (n = 5).

CONCLUSIONThe results showed that this method was reliable and accurate, and can be used for quality control of Fufang Guilu granule.

Betaine ; analysis ; Chromatography, High Pressure Liquid ; methods ; Codonopsis ; chemistry ; Drug Combinations ; Drugs, Chinese Herbal ; chemistry ; Lycium ; chemistry ; Plants, Medicinal ; chemistry ; Quality Control ; Reproducibility of Results

OBJECTIVETo investigate the polymorphism distribution of vitamin D receptor (VDR) gene Fok I in Mongolian population of China.

METHODSPolymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method was used to analyze three genotypes FF, Ff and ff in the start codon of VDR gene (Fok I) in unrelated normal healthy Mongolian individuals of China.

RESULTSIn the population, we obtained the allelic frequencies of 57% and 43% for (F) and (f) allele and the percentage of genotypes FF, Ff and ff to be 31%, 52%, and 17% respectively.

CONCLUSIONThe polymorphism frequency and distribution of this VDR gene Fok I in Mongolian population of China exhibit its own characteristics.

Adult ; Aged ; Asian Continental Ancestry Group ; genetics ; China ; ethnology ; Female ; Genotype ; Humans ; Male ; Middle Aged ; Polymorphism, Genetic ; Population ; Population Groups ; genetics ; Receptors, Calcitriol ; genetics

OBJECTIVETo establish strategies for preventing graft versus host disease (GVHD) and reducing treatment associated morbidity while preserving graft versus leukemia (GVL) effect in nonmyeloablative allogeneic bone marrow transplantation (allo-BMT), with or without donor lymphocyte infusion (DLI) after BMT.

METHODS3 x 10(7) bone marrow cells mixed with 1 x 10(7) spleen cells from the same BALB/c mouse were transplanted into the nonablative irradiated inbred 615 mouse which received a single subcutaneous injection of 1 x 10(6) L615 leukemia cells three days before. The experiments were designed as follows (ten mice in each group): myeloablative BMT control group (group A), nonmyeloablative conditioning without BMT group (group B), nonmyeloablative BMT group (group C), and nonmyeloablative BMT + DLI group (group D). GVL effects were assessed by survival time, white blood cell count and L615 cells in peripheral blood and histologic changes. GVHD was assessed by signs of weight loss, ruffled fur, diarrhea and histologic changes of skin, liver and small intestines. Chimerism was detected by cytogenetic analysis and PCR technique.

RESULTSThe survival time of group A, B, C and D was (20.3 +/- 13.4), (15.9 +/- 1.1), (21.6 +/- 1.7) and (37.8 +/- 2.0) days, respectively, being no significant difference between group A and group C (P > 0.05). The survival time of group C was longer than that of group B (P < 0.01). And among group B, C and D, group D had the longest survival time (P < 0.01). GVHD signs and histologic changes were observed in 60% of control group mice at + 14 day, but none of group C and group D. 40% of mice in group A died of treatment associated morbidity within two weeks, but none in group C and group D. Allogeneic chimerism was kept in group A, but excluded gradually in group C.

CONCLUSIONGVL effect seems preserved in nonmyeloablative BMT mice, but weaker than that in myeloablative BMT mice. GVL effect seems to be enhanced by DLI after nonmyeloablative BMT. GVHD and transplantation associated morbidity seems to be reduced in nonmyeloablative BMT.

Animals ; Bone Marrow Transplantation ; immunology ; methods ; Combined Modality Therapy ; Female ; Graft vs Host Disease ; prevention & control ; Graft vs Leukemia Effect ; Leukemia, Experimental ; therapy ; Leukemia, Lymphoid ; therapy ; Lymphocyte Transfusion ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Inbred Strains ; Transplantation Conditioning ; methods ; Transplantation, Heterologous

AIM: Accommodation is one of the most important functions of human eye, while its mechanism is still under discussion. This paper aimed to study accommodation mechanism with numerical simulation.METHODS: A simulation model was constructed to study the mechanism of accommodation based on the experimental data derived from published resources. The displacement and pressure are applied on the model to study the deformation of lens during accommodating.RESULTS: The simulation showed that, as the eye was accommodating, the thickness of the lens increased linearly,and the lens diameter decreased linearly. The optical power of the lens increased as the accommodation increased. This result was accord with the public facts in accommodation.Furthermore, the pressure was found to have a great influence on the shape of the lens and the optical power. The lens became thinner and flatter as the pressure increased and the pressure caused a remarkable increase of lens' optical power.CONCLUSION: The outcome of this paper is consistent with the Helmholtz's hypothesis on accommodation to some extent. The analytical model presented in this paper can be used in the theoretical study of the accommodation mechanism of the human lens.

OBJECTIVETo study the anatomy of lateral crural skin flap nourished by cutaneous branches of peroneal artery and its clinical application as vascularized skin flap transfer.

METHODSIn 20 cadavers specimen with 40 lower limbs, the cutaneous branches of the peroneal artery were dissected and their measurements were recorded. In the other 30 adult legs, their perforating points of the cutaneous arteries of peroneal artery were detected with supersonic Doppler flow meter. With the aid of anatomic and supersonic Doppler flow meter study, vascularized transfer of lateral crural skin flap pedicled by cutaneous branches of peroneal artery were successfully performed in 21 clinical cases.

RESULTSIn altogether 40 legs studied, 140 cutaneous branches were found. One to seven branches were found on one specimen, the average was 3.5 branches, in one leg was a high perforating skin branch. The perforating points of the cutaneous branches were mostly (76% cases) appeared within 7 - 21 cm length below the protruding point of head of fibula. The external diameter of the thickest cutaneous branch of each leg was (1 .4 - 2.9) mm, (1.8 +/- 0.4) mm, while the external diameters of two vena concomitants were (3.0 +/- 0.5) mm and (2.4 +/- 0.4) mm. 145 artery perforating points in 30 legs were detected by Doppler, with an average points of 4.8. The skin flaps taken in the 21 clinical cases were 5.0 cm x 3.5 cm - 28 cm x 11 cm in size. All the transferred free flaps survived uneventfully.

CONCLUSIONSThe lateral crural skin flap is nourished by a variable number of cutaneous branches of peroneal artery. The main branch can meet the demand of microvascular anastomosis. The free transfer of lateral crural flap by anastomosis of cutaneous branch of peroneal artery is superior to lateral skin flap transfer by anastomosis of main trunk of peroneal artery with the merit of simple procedure, minimal trauma and more physiological circulation established.

Adult ; Aged ; Arteries ; anatomy & histology ; surgery ; Humans ; Leg ; blood supply ; Middle Aged ; Skin ; anatomy & histology ; blood supply ; Skin Transplantation ; methods ; Surgical Flaps ; blood supply ; Young Adult

AIMTo investigate the effect of baicalin on the hippocampal neuronal apoptosis and the expression of HSP70 in rats with focal brain ischemia-reperfusion injury.

METHODSOne hundred and twenty male Wistar rats were randomly divided into six groups:sham operated group, ischemia-reperfusion group, nimodipine group and three baicalin groups,to which baicalin was administered at doses of 50, 100 and 200 mg x kg(-1), separately. The models of focal brain ischemia-reperfusion injury induced by middle cerebral artery occlusion (MCAO) were used in this study. HE stain was used to observe the pathological changes. Flow cytometry (FCM) was used for determination of neuronal apoptosis. HSP70 protein expression of the neurons was detected with immunohistochemistry. Reverse transcription polymerase chain reaction (RT-PCR) was used to detect the expression of the mRNA level of HSP70.

RESULTSBaicalin can significantly relieve the pathological changes and inhibit apoptosis in hippocampus CA1 area, and at the same time increase the expression of HSP70 and HSP70 mRNA.

CONCLUSIONBaicalin can relieve brain damage induced by focal brain ischemia-reperfusion in rats, which may be related to inhibiting the process of the neuronal apoptosis. The mechanism of antiapoptosis effect of baicalin may be related to the promotion of transcription of HSP70 mRNA and increasing the expression of the protein.

Animals ; Apoptosis ; drug effects ; Flavonoids ; isolation & purification ; pharmacology ; Flow Cytometry ; HSP70 Heat-Shock Proteins ; biosynthesis ; genetics ; Hippocampus ; drug effects ; metabolism ; pathology ; Infarction, Middle Cerebral Artery ; complications ; Male ; Neurons ; drug effects ; metabolism ; pathology ; Neuroprotective Agents ; isolation & purification ; pharmacology ; Plants, Medicinal ; chemistry ; Pyramidal Cells ; drug effects ; metabolism ; pathology ; RNA, Messenger ; biosynthesis ; genetics ; Random Allocation ; Rats ; Rats, Wistar ; Reperfusion Injury ; etiology ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Scutellaria ; chemistry

BACKGROUNDThe application of pulmonary valved conduit to reconstruct the continuity between right ventricles and pulmonary artery is one of the major surgeries. This study aimed to establish an in vivo model of in situ implantation using pulmonary valved conduit in large animals under off-pump condition to validate the long-term effects of artificial pulmonary valved conduit.

METHODSDomesticate juvenile male sheep and tissue-engineered porcine pulmonary valved conduit were used for the experiment: 30 sheep, weighing (15 ± 3) kg (range 13 to 17 kg) were randomly divided into two groups which were all operated under general anesthesia by off-pump surgery (group 1) and left thoracotomy (group 2). Two different off-pump surgical methods were used to perform cannulation in sheep pulmonary artery to replace part of sheep pulmonary artery with pulmonary valved conduit which will work together with sheep pulmonary artery and valves. During the experiments, animal survival, complication rates, operating time and blood loss were recorded to compare the results between groups and to establish a surgical method with minimal invasion, simplicity, safety, and high success rates.

RESULTSIn group 1, a total of 15 cases of surgeries were performed, in which two sheep died; the operative mortality was 13.3% (2/15). In group 2, a total of 15 cases of surgeries were performed, and the surgical mortality rate was 0 (0/15). The operation time and blood loss in group 2 was significantly better than that in group 1. The postoperative echocardiograms showed that, after the surgeries by these two methods, the blood flows were normal, and the valves can open and close freely. Autopsy after 6 months showed that the inner wall and the valves of pulmonary valved conduit were smooth with no thrombus formation.

CONCLUSIONThese two off-pump methods are feasible and safe with fewer traumas; but the second method is better and particularly suitable for the establishment of a juvenile animal model.

Animals ; Heart Valve Prosthesis ; Male ; Pulmonary Valve ; Sheep ; Swine ; Tissue Engineering

OBJECTIVETo identify gene expression patterns in distinct stages of intestinal-type gastric cancer(GC).

METHODSGene expression patterns of distinct stages of intestinal-type GC samples from 3 patients were compared with cDNA microarray, which contained 576 genes. There were 506 target genes, which included 51 genes identified from our previous experiment with suppression subtractive hybridization(SSH) and other 455 genes chosen for their important roles in cancers. Hierarchical clustering was performed to clarify genes in association with distinct stages of GC.

RESULTSOne hundred and eighty-one differentially expressed genes with average Cy5:Cy3 ratios higher than 2.0 or lower than 0.5 in at least one stage of GC were identified by cDNA microarray. Among them, 48 genes were up-regulated and 133 down-regulated. Hierarchical clustering analysis separated the differentially expressed genes in different stages of GC into 5 main characteristic groups. Some important differentially expressed genes in different stages of GC were identified, such as SEC23IP, LIPF, ES(BQ291520), SLC5A1, PG(encoding similar to pepsin A precursor), CXCR4, DICER1, SH3GL2, and IGF2R.

CONCLUSIONThe differentially expressed gene patterns and some important genes were identified, which might be useful in further study on carcinogenesis, progression and metastasis of intestinal-type GC.

DNA, Neoplasm ; analysis ; Gene Expression ; Gene Expression Profiling ; Gene Expression Regulation, Neoplastic ; Gene Library ; Humans ; Microarray Analysis ; Nucleic Acid Hybridization ; Oligonucleotide Array Sequence Analysis ; Stomach Neoplasms ; genetics ; metabolism ; Transcription, Genetic

OBJECTIVETo construct cDNA subtracted libraries from gastric dysplasia and further screen differentially expressed genes.

METHODSRelatively pure dysplasia and normal tissue were procured by manual microdissection, and amplified by cDNA-PCR, which was used to carry on for suppression subtractive hybridization (SSH). Subtracted cDNA fragments were linked with vector, cloned, screened, sequenced, and made homologous search. Differentially expressed fragments were verified by dot hybridization.

RESULTSTwo subtracted cDNA libraries were constructed. Among 26 sequenced clones, 15 fragments corresponded to known genes, 3 fragments were known EST and 8 fragments were unknown EST (GenBank BQ164614-BQ164616, BQ291516-BQ291520). Fifteen fragments were verified to be differentially expressed in gastric dysplasia.

CONCLUSIONSSubtracted cDNA libraries from gastric dysplasia are constructed using combination of microdissection-cDNA PCR and SSH setup in our laboratory. Some fragments have been screened and verified to help to search for novel associated genes with gastric carcinogenesis.

Cloning, Molecular ; DNA, Complementary ; genetics ; Gene Expression Profiling ; Gene Expression Regulation, Neoplastic ; Gene Library ; Humans ; Microdissection ; Nucleic Acid Hybridization ; methods ; Polymerase Chain Reaction ; Precancerous Conditions ; genetics ; pathology ; Sequence Analysis, DNA ; Stomach Neoplasms ; genetics ; pathology

OBJECTIVETo screen and analyze the important associated genes in different stages of gastric cancer.

METHODSUsing suppression subtractive hybridization (SSH) to screen differentially expressed genes; detecting the expression of genes in different stages of gastric cancer with dot blot hybridization; and verifying the results with semi-quantitative reverse transcriptase-polymerase chain reaction(RT-PCR).

RESULTSTwenty-six differentially expressed gene fragments were obtained by means of SSH. Among them,24 were known genes, 1 was a new expressed sequence tags(EST), and 1 was a hypothetical gene. The results of dot blot hybridization demonstrated that the expressions of Annexin A2, RPS29, RPS12 etc. in dysplasia were higher than those in normal mucosa; the expressions of RPS12 etc.in early cancer were higher than those in normal mucosa;the expressions of cytochromosome C oxidase II, ferritin light chain, RPS12 etc. in advanced gastric cancer and lymph node metastases were consistently higher than those in normal mucosa. The expression of proteasome 26S subunit gene in advanced gastric cancer was higher than that in normal mucosa. The expression of RPS12 was consistently higher in different stages of gastric cancer. It was demonstrated by RT-PCR that the expression of RPS12 in gastric cancer was higher than that in normal mucosa.

CONCLUSIONThe authors have identified some important genes that might be involved in the carcinogenesis and progression of gastric cancer, and RPS12 may play more important roles in gastric cancer.

Gene Expression Profiling ; methods ; Gene Expression Regulation, Neoplastic ; Genetic Testing ; methods ; Humans ; Nucleic Acid Hybridization ; methods ; Reverse Transcriptase Polymerase Chain Reaction ; Stomach Neoplasms ; diagnosis ; genetics