Chronic Intracerebroventricular Infusion of Metformin Inhibits Salt-Sensitive Hypertension via Attenuation of Oxidative Stress and Neurohormonal Excitation in Rat Paraventricular Nucleus / 神经科学通报·英文版

Metformin (MET), an antidiabetic agent, also has antioxidative effects in metabolic-related hypertension. This study was designed to determine whether MET has anti-hypertensive effects in salt-sensitive hypertensive rats by inhibiting oxidative stress in the hypothalamic paraventricular nucleus (PVN). Salt-sensitive rats received a high-salt (HS) diet to induce hypertension, or a normal-salt (NS) diet as control. At the same time, they received intracerebroventricular (ICV) infusion of MET or vehicle for 6 weeks. We found that HS rats had higher oxidative stress levels and mean arterial pressure (MAP) than NS rats. ICV infusion of MET attenuated MAP and reduced plasma norepinephrine levels in HS rats. It also decreased reactive oxygen species and the expression of subunits of NAD(P)H oxidase, improved the superoxide dismutase activity, reduced components of the renin-angiotensin system, and altered neurotransmitters in the PVN. Our findings suggest that central MET administration lowers MAP in salt-sensitive hypertension via attenuating oxidative stress, inhibiting the renin-angiotensin system, and restoring the balance between excitatory and inhibitory neurotransmitters in the PVN.

The change of COX-2 expression in rats with stress urinary incontinence and its significance / 医学研究生学报

Objective　There are a few researches on the mechanism of stress urinary incontinence (SUI). The article aimed to examine the changes of COX-2 expression in the urethra, vagina and urethral smooth muscle of SUI rat mode to evaluate the effect of estrogen on COX-2 expression. Methods　Sixty unbearing healthy female SD rats and fifteen male SD rats were gathered for spontaneous delivery. SUI rat models were constructed using expanded vagina, expanded vagina + ovariectomy respectively after delivery, which were expanded vagina group and expanded vagina + ovariectomy group. Six successfully modeled rats were chosen for the follow-up experiment. SD rats modeled after normal pregnancy were the control group. Sneezing experiment and urodynamic examination were used to examine the maximum bladder capacity (MBC) and abdominal leak point pressure (ALPP). Fluorescent quantitative PCR and western blot were applied to detect the expressions of COX-2 mRNA and protein, and immunohistochemistry was used to detect the expressions of COX-2 in urethra, vagina and urethral smooth muscle. Results Compared with control group, ALPP in two experimental groups were significantly decreased, among which ALPP in expanded vagina + ovariectomy group was significantly decreased in comparison to expanded vagina group(P<0.05). Compared with control group, the expressions of COX-2 mRNA and protein in expanded vagina group and expanded vagina+ovariectomy group were significantly higher, among which the figures in expanded vagina+ovariectomy group were significantly higher than those in expanded vagina group(P<0.05). The result of immunohistochemistry showed staining intensity integral expression of COX-2 in vaginal tissues of control group, expanded vagina group and expanded vagina+ovariectomy group were 0.50±0.54, 5.55±0.54, 9.33±0.81, so differences between any two groups were of statistical significance(P<0.05); staining intensity integral expression of COX-2 in urethral smooth muscle of control group, expanded vagina group and expanded vagina+ovariectomy group were 0.66±0.51, 5.33±0.51, 8.50±0.54, so differences between any two groups were of statistical significance (P<0.05). Conclusion The expression of COX-2 was related to the mechanism of SUI. The decrease of estrogen may increase the expression of COX-2 in SUI rats, which supports the treatment of SUI.

Acanthopanax senticosus Protects Structure and Function of Mesencephalic Mitochondria in A Mouse Model of Parkinson's Disease / 中国结合医学杂志

<p><b>OBJECTIVE</b>To investigate the neuro-protective effects of Acanthopanax senticosus Harms (EAS) on mesencephalic mitochondria and the mechanism of action, using a mouse model of Parkinson's disease (PD).</p><p><b>METHODS</b>The chemical fingerprint analysis of the extract of Acanthopanax senticosus Harms (EAS) was performed using the ultra performance liquid chromatograph and time of flight mass spectrometry. Thirty mice were randomly divided into the control group, the MPTP model group, and the EAS treated group with MPTP (MPTP+EAS group, 10 in each group). The MPTP model group and the MPTP+EAS group received MPTP-HCl (30 mg/kg i.p) once a day for 5 days. The control group received an equal volume of saline (20 mL/kg i.p) once a day for 5 days. Induced by 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine hydrochloride daily (MPTP-HCl, 30 mg/kg) for 5 days, the PD mice were treated with EAS at 45.5 mg/kg daily for 20 days. The behavioral testing of mice was carried out using the pole-climbing test. The integrity and functions of neurons were examined in mesencephalic mitochondria in a PD mouse model, including nicotinamide adenine dinucleotide dehydrogenase ubiquinone flavoprotein 2 (NDUFV2), mitochondrially encoded nicotinamide adenine dinucleotide dehydrogenase 1 (MT-ND1), succinate dehydrogenase complex subunit A (SDHA), and succinate dehydrogenase cytochrome b560 subunit (SDHC).</p><p><b>RESULTS</b>After treatment with EAS, the behavioral changes induced by MPTP were attenuated significantly (P<0.05). EAS protected the mesencephalic mitochondria from swelling and attenuated the decreases in their membrane potential (both P<0.05), which was supported by an ultra-structural level analysis. The changes in reactive oxygen species (ROS), malonic dialdehyde (MDA), oxidative phosphorylation (OXPHOS) system 4 subunits levels and PD-related proteins expressions (parkin, Pink1, DJ-1, α-synuclein, and Lrrk2) reverted to near normal levels (all P<0.05), based on the results of immune-histological and Western blotting observations.</p><p><b>CONCLUSIONS</b>The neuro-protective effects of EAS are linked to protecting mice against MPTP-induced mitochondrial dysfunction and structural damage. Therefore, EAS is a promising candidate for the prevention or treatment of mitochondrial neurodegenerative disorders, such as PD.</p>

Cordblood-Based High-Throughput Screening for Deafness Gene of 646 Newborns in Jinan Area of China

OBJECTIVES: Infants with slight/mild or late-onset hearing impairment might be missed in universal newborn hearing screening (UNHS). We identified the mutation hot spot of common deaf gene in the newborns in Jinan area population by screening the mutation spot with neonate cord blood, in order to make clear whether the neonate cord blood for screening is feasible. METHODS: Six hundred and forty-six newborns were subjected to both UNHS and genetic screening for deafness by using neonate cord blood. The newborn genetic screening targeted four deafness-associated genes, which were commonly found in the Chinese population including gap junction beta-2 protein (GJB2), gap junction beta-3 protein (GJB3), solute carrier family 26 member 4 (SLC26A4), and mtDNA 12S rRNA. The most common 20 spot mutations in 4 deaf genes were detected by MassARRAY iPLEX platform and mitochondrial 12S rRNA A1555G and C1494T mutations were sequenced using Sanger sequencing. RESULTS: Among the 646 newborns, 635 cases passed the UNHS and the other 11 cases (1.7%) did not. Of the 11 failures, two cases were found to carry homozygous GJB2 p.R143W pathogenic mutation, one case was found to have heterozygous GJB2 235delC mutation, and another one case carried heterozygous GJB3 p.R180X pathogenic mutation. Six hundred and thirty-five babies passed the newborn hearing screening, in which 25 babies were identified to carry pathogenic mutations, including 12 heterozygotes (1.9%) for GJB2 235delC, eight heterozygotes (1.3%) for SLC26A4 IVS7-2A>G, one heterozygote (0.2%) for p.R409H, two homozygotes (0.3%) for m.1494C>T, and two homozygotes (0.3%) for m.1555A>G. CONCLUSION: Newborn genetic screening through the umbilical cord blood for common deafness-associated mutations may identify carriers sensitive to aminoglycoside antibiotic, and can effectively prevent or delay hearing loss occurs.

Research Review on TCM Preventing and Treating Parkinson's Disease by Repairing Brain / 中国中医药信息杂志

Parkinson’s disease (PD) is a common degenerative disease of central nervous system. Brain mitochondrial dysfunction and structural damage are the important pathogeny of PD. At present, many of Chinese herbal compounds, herbal medicines, and TCM active ingredients are used to prevent and treat PD. The main mechanisms of these medicines are involved in the protection of mitochondrial structure, anti-oxidative stress, anti-calcium dysregulation, mitigation of excitotoxicity, and anti-apoptosis, etc., which also play a comprehensive role through multi-link, multi-level, and multi-target. Through looking up the recent representative literature, the experimental results of Chinese herbal compounds and TCM active ingredients preventing and treating PD through repairing brain mitochondrial structure and function were analyzed and inducted. Many of Chinese herbal compounds and TCM active ingredients were proved to have good effects on PD.

Social participation and needs for rehabilitation services of the disabled in Guangdong Province HUANG / 中华物理医学与康复杂志

Objective To explore the relationship between social participation and needs for rehabilitation of the disabled in Guangdong Province and to make a proposal for developing the rehabilitation strategies. Meth-ods The data of the Second National Sample Survey of Disabled Persons in Guangdong Province was used in this study. Ranked data analysis was made with the sub-items of the social participation assessment and the main needs of the disabled individuals. Results Significantly statistical differences were revealed with regard to the constitu-ent ratio of needs for rehabilitation services among people with different degrees of difficulties in social participation caused by hearing and visual impairments as well as physical and mental disabilities. No significant difference was found in terms of the constituent ratio of rehabilitation needs among those with difficulties in speech and those with psychiatric diseases. The major rehabilitation needs focused on medical service, assistive apparatus support and functional trainings. Conclusions The rehabilitation needs were different among different categories of disabled persons. Rehabilitation services should be provided accordingly.

Study on the prevalence of metabolic syndrome among the Kazakh population in Xinjiang / 中华流行病学杂志

Objective To analyze the prevalence of metabolic syndrome (MS) in Kazakh population, using the NCEP-ATP Ⅲ, CDS, IDF MS standards. Methods Questionnaire-based survey,physical examination and blood testing were conducted according to cluster random samplings in Kazakh residents in Xinjiang. 2745 samples were collected and diagnosed by NCEP-ATP Ⅲ, CDS,IDF standards to analyze the prevalence, with the distribution of its main components of MS, among the Kazakhs population. Results The prevalence rates of MS diagnosed by NCEP-ATP Ⅲ, CDS,IDF standards were 18.5%, 14.2% and 26.6%, while they became 14.2%, 10.9% and 20.1% after standardized by age. The prevalence of MS diagnosed by NCEP-ATP Ⅲ and IDF standard in males were higher than in females, while CDS was in the opposite situtation. The prevalence of MS by these three standards increased with age. Among all the main components of MS diagnosed after these three standardization process, the prevalence of obesity, blood pressure rising and the abnormity of HDL-C were rather high. The prevalence of MS main components ≥1, ≥2, ≥3, ≥4, 5 ranked the highest compared to the lowest as to the IDF, ATP Ⅲ ' and CDS diagnostic. standards Conclusion The prevalence rates and gender distribution of MS diagnosed by different standards among Kazakhs were different. The prevalence of IDF standard was the highest, with the IDF standard better than the others in early identifying the risk factors of cardiovascular disease.

Association of lipoprotein lipase gene Hind Ⅲ and S447X polymorphisms in metabolic syndrome patients among Kazakh and Han ethnics from Xinjiang / 中华流行病学杂志

Objective To investigate the association of lipoprotein lipase gene Hind Ⅲ and S447X polymorphisms with metabolic syndrome among Kazakh and Han ethnicities in Xinjiang.Methods PCR-RFLP was used to detect 802 subjects' lipoprotein lipase Hind Ⅲ and S447Xgenotypes (including 201 controls and 200 metabolic syndrome patients in Kazakh and Han ethnicities, respectively). Results (1) Frequencies of H + H-/H-H- genotype (32.50% vs.47.76%), H- allele( 18.00% vs. 28.86%), SX/XX genotype (8.00% vs. 22.39%) and X allele (4.00%vs. 12.44% ) for metabolic syndrome in Hah ethnicity were all significantly lower than those in controls (P＜ 0.01 ). (2) The frequencies of H + H-/H-H- genotype (33.50% vs. 46.80% ), H- allele (22.00% vs. 28.60%), SX/XX genotype (10.50% vs. 22.90%) and X allele (5.50% vs. 12.44% ) in patients with metabolic syndrome in Kazakh were all significantly lower than those for controls (P＜0.01). (3) The frequencies of lipoprotein lipase gene Hind Ⅲ and S447X genotypes and alleles in Kazakh were not significantly different from Han (all P＞0.05). (4)The levels of waist circumference, systolic blood pressure, diastolic blood pressure, triglyceride and FPG in H + H-/H-H- and SX/XX genotype were significantly lower than those in H + H + and SS genotype.HDL-C was significantly higher than that in H + H + and SS genotype (P＜0.05). (5) The frequencies of H + H + and SS genotype increased along with the increase in number of metabolic syndrome component. Conclusion The lipoprotein lipase gene Hind Ⅲ and S447X polymorphisms were associated with metabolic syndrome risk in Kazakh, and H + H-/H-H- genotype, H- allele, SX/XX genotype and X allele might have served as protective factors of metabolic syndrome. H + H-/H-H- and SX/XX genotype seemed to have had beneficial effects for all the metabolic syndrome components, and the frequencies of H + H + and SS genotype were increasing along with the increase of number in the metabolic syndrome components.

Experiment of oral mucosa epithelial cells cultured on small intestinal submucosa in vitro / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To explore an effective method to culture oral mucosa epithelial cells (OMECs) of canine in vitro, and to observe the biological characteristics of OMECs growing on small intestinal submucosa (SIS) in order to provide the experimental basis for epithelium tissue engineering.</p><p><b>METHODS</b>The primary OMECs were cultivated with DKSFM (defined keratinocyte serum free medium) containing 6% fetal bovine serum (FBS). The morphological characteristics and the growth curve of OMECs were observed. The expressions of OMECs marker (CK19) were examined by immunocytochemistry. The 2nd passage of OMECs were seeded on SIS, OMECs co-cultured with SIS were observed by hematoxylin-eosin staining, immunohistochemical staining, and scanning electron microscope (SEM).</p><p><b>RESULTS</b>OMECs were grown well in DKSFM. Immunohistochemical staining of the 2nd passage cultured canine OMECs with broadly reacting anti-cytokeratin anyibodies (CK19) was positive. OMECs formed a single layer on the surface of SIS, and eight days later the cells were polygong and arranged like slabstone.</p><p><b>CONCLUSION</b>Culture of canine OMECs in DKSFM containing 6% FBS is a simple and feasible method. SIS has good biocompatibility, it is a kind of good bioscafold in the tissue-engineered epithelium.</p>

Comparative study on biochemical characteristic of Yersinia pestis from some natural focuses of plague in Yunnan Province / 中国地方病学杂志

Objective To compare the difference of biochemical characteristics and virulent Pst Ⅰ of Yersinia pestis strains in traditional focuses of plague in Yunna Province and in the new focuses of plague in Yulong County. Methods The identification data of biochemical characteristics(Rhamnose, Glycerol, Maltose, L-Arabina and Melibiose fermentation) and virulence factor(Pst Ⅰ) from different focuses of plague in Yunna Province were Retrospectively collected by tube test followed by the analysis using statistics software SAS 8.0 by Fisher exact probability of disordered two-way R × C table χ2 test. Results Among 48 strains of Yersinia pestis from hantaan type plague focus, 1 strain fermented L-maltose, 48 strains fermented Glycerol. Among 165 strains of Yersinia pestis from the Soul type plague focus, 1 strain did not ferment L-maltose, only one of them fermented Glycerol. 1 strain from the Soul type plague focus was confirmed to have mutation, for the test of nitrate reduction reaction was negative. All 5 strains of Yersinia pestis from the new focuses of plague in Yulong County fermented L-maltose and Glycerol. The statistical result showed that the differences in L-maltose and Glycerol fermentation of Yersinia pestis from different natural focuses of plague in Yunnan Province were statistically siguificant (P < 0.01). The differences of other biochemical characteristics and Pst Ⅰ were not statistically significant (P > 0.01). Conclusions Biochemical characteristics of Yersinia pestis from the hantaan type plague focus and the Soul type plague focus in Yunnan province are overlapping. Biochemical characteristics of Yersinia pestis from the new focuses of plague in Yulong County are different from those tradition focuses of plague in Yunna Province but share similarities to those from Unquiculatus focuses in North Tibet.

The possible role of Myosin light chain in myoblast proliferation

Skeletal muscles have the potential to regenerate by activation of quiescent satellite cells, however, the molecular signature that governs satellite cells during muscle regeneration is not well defined. Myosin light chains (Myls) are sarcomere-related proteins as traditional regulator of muscle contraction. In this report, we studied the possible role of Myl in the proliferation of skeletal muscle-derived myoblasts. Compared to diaphragm-derived myoblasts, the extraocular muscle-derived myoblasts with lower levels of Myl proliferated faster, maintained a longer proliferation phase, and formed more final myotubes. It was found that blockading Myl with anti-Myl antibody or knockdown of Myll by siRNA targeted against Myll could enhance the myoblast proliferation and delay the differentiation of myoblasts. Our results suggested that Myl, likely Myll, can negatively affect myoblast proliferation by facilitating myoblast withdrawal from cell cycle and differentiation.

Analysis of the survey result of the coal-burning endemic fluorosis in Hongya County of Sichuan Province in 2006 / 中国地方病学杂志

Objective To investigate the current status of coal-burning endemic fluorosis and the fluoride content in foods and drinking water in Hongya County,Sichuan Province.Methods Dental fluorosis and urinary fluoride were suveyed in children of 8～12 years old in two schools which repectively located in Gaomiao and Wawushan Town.The adults above 20 years old underwent clinical examination.At the same time,fifty adults above 20 years old in Garden Village were chosen to take forearm and calf X-ray films to find out the evidence of skeletal fluorosis.The content of fluoride in food such as bacon,corn,dry capsicum in Sanxing Village in Gaomiao Town and Futian Village in Wawushan Town as well as drinking water in five families in Sanxing Village were determined.Results The dental fluorosis rate of children was 40.76%(161/395),the dental fluorosis index was 0.86 in Gaomiao Town.The dental fluorosis rate of children was 14.36%(82/571),the dental fluorosis index was 0.31 in Wawushan Town.The medium value of the urine fluoride was 0.81 mg/L.ranged 0.16～3.89 mg/L.The positive rate oi the clinical examination of skeletal fluorosis was 5.27%(27/512),the X-rays detective rate was 4.00%(2/50).The medium value in bacon,corn,dry capsicum were 6.00,0.64,1.49 mg/kg.The averaged content of the fluoride in drinking water(0.14±0.06)mg/L of local household was within the eligible limitation.Conclusions It is currently a mild endemic disease in Hongya Country,its incidence is reduced apparently,pathogenetic environmental fluoride content is reduced.The main source of fluoride is from the preserved ham contaminated with fluoride,which is epidemiologically significant in endemic area of Hongya County.Defluoriding countermeasures should be taken in the endemic areas.

Seroepidemiological survey of F1 antibody of indicator animals in wild rodents loci of plague in YunnanProvince / 中国地方病学杂志

Objective To study on the effect of indicator animals in plague surveillance throngh detecting F1 antibody against Yersiniapestis(Y.pestis)in indicator animals in wild rat plague foci,provide scientific evidence for plague control and determining the range of epidemic area.Methods According to investigation scheme of wild rodents plagne foci in Yunnan Province,indicator animals Canis familiarils and Felis catu(C.familiarils and F.catus)related to the plague were investigated in 75 villages,14 township and 10 counties around Yulong County,and living rodents were captured by cage,sera of indicator animals and rodents relevant to plague were simultaneously collected and detected for F1 antibody against Y.pestis using indirect hemagglutination(IHA).Results Seropositivity rate of indicator animals were 6.76%(202/2987),being 24.69% in C.familiaris and 24.69% in F.catus,there were statistical significance(X2＝87.32,P＜0.01)between C familiaris and F catus,the latter beingmore than the former.But F1 antibody of rodents sera were not detected,its seropositivity rate was zero.there was a statistical significance(P＜0.01)between indicator animals and rodents.Conclusions Through serocpidemiological survey of indicator animals,new wild rat plague natural focus has been confirmed in YuLong County and Gucheng District in LiJiang City,therefore,serocpidemiological surveillance of indicator animals is very important for plague control and prevention.

Current understanding of dystrophin-related muscular dystrophy and therapeutic challenges ahead / 中华医学杂志(英文版)

<p><b>OBJECTIVE</b>To review the recent research progress in dystrophin-related muscular dystrophy includes X-linked hereditary Duchenne and Becker muscular dystrophies (DMD and BMD).</p><p><b>DATA SOURCES</b>Information included in this article was identified by searches of PUBMED and other online resources using the key terms DMD, dystrophin, mutations, animal models, pathophysiology, gene expression, stem cells, gene therapy, cell therapy, and pharmacological. Study selection Mainly original milestone articles and timely reviews written by major pioneer investigators of the field were selected.</p><p><b>RESULTS</b>The key issues related to the genetic basis and pathophysiological factors of the diseases were critically addressed. The availabilities and advantages of various animal models for the diseases were described. Major molecular and cellular therapeutic approaches were also discussed, many of which have indeed exhibited some success in pre-clinical studies but at the same time encountered a number of technical hurdles, including the efficient and systemic delivery of a functional gene and myogenic precursor/stem cells to repair genetic defects.</p><p><b>CONCLUSIONS</b>Further understanding of pathophysiological mechanisms at molecular levels and regenerative properties of myogenic precursor/stem cells will promote the development of multiple therapeutic strategies. The combined use of multiple strategies may represent the major challenge as well as the greatest hope for the therapy of these diseases in coming years.</p>

The experiment of porcine keratinocytes cultured on porcine small intestinal submucosa in vitro / 中华整形外科杂志

<p><b>OBJECTIVE</b>To investigate the feasibility of acellular porcine small intestinal submucosa (SIS as bioscaffold of tissue engineering skin.</p><p><b>METHODS</b>The second passage keratinocytes were seeded on SIS, after seeded for 11, 13, 15, 17 days, the keratinocytes/SIS composites were observed by dye directly, histopathology, immunohistochemical studies with monoclonal antibodies against laminin and transmission electron microscope (TEM).</p><p><b>RESULTS</b>At eleventh day, keratinocytes were growth very well on the surface of SIS, there are 2-3 cell layers on partial of the SIS surface, the continued expression of laminin can be detected between the keratinocytes and the surface of SIS. After 13, 15, 17 days this stratified structure increased, cells contact more closely, the tonofibrils in cells, desmosome between cells and the basal membrane between the keratinocytes and the surface of SIS can be seen with TEM.</p><p><b>CONCLUSIONS</b>SIS is a kind of good bioscaffold in the culture of porcine keratinocytes in vitro.</p>

A Study in Construction of Short Hairpin Small Interfering RNA Expression Vector Target Lectin Like Oxidized Low Density Lipoprotein Receptor-1 Gene and Its Effect on Foam Cells / 中国生物工程杂志

Aim:To construct the short hairpin small interfering RNA(shRNA) eukaryotic expression vector specific to mouse lectin like oxidized low density lipoprotein receptor 1(LOX-1) gene and to observe its silencing effect on LOX-1 in RAW264.7 cells.Methods:(1)The pLOX-1-shRNA expression vector was constructed by gene recombination,Then transfected into the cultured RAW264.7 cells.At 48 h after Transfection,the expression of LOX-1 mRNA in RAW264.7 cells were determined by semi-quantitative RT-PCR,the expression of LOX-1 proteins examined by Western blot.(2) Oil Red O Dyeing experiment was used to show the cellular lipid droplets in lipid-loaded cells.The method of cholesterol oxidase analysis was performed to determine the content of cellular cholesterol.The ability of uptake Dil-ox-LDL in RAW264.7 cells was assayed by fluorescence microscopy.Results: pLOX-1-shRNA expression vector was successfully constructed.Transfection of pLOX-1-shRNA expression vector into RAW264.7 cells down regulaled the expression level of LOX-1 gene,as compared with the control group,transfection of the RAW264.7 cells with LOX 1-shRNA led to a remarkable reduction of the number macrophages transformed into foam cell,and could suppress the uptake of ox-LDL.Conclusion:The pLOX-1-shRNA expression vector can inhibit the expression of LOX 1 in RAW264.7 cells and the transformation of the macrophages into foam,which may he beneficial in searching new gene therapy of atherosclerosis.

Culture of osteoblasts on bio-derived bones / 中华创伤杂志(英文版)

<p><b>OBJECTIVE</b>To study the effect of bio-derived bones, as substitutes of autogenous bone grafts and demineralized cadaver bones, on the attachment, spreading and proliferation of isolated osteoblasts.</p><p><b>METHODS</b>Osteoblasts were isolated from the calvaria of a fetal rabbit through sequential collagenase digestion. In the attachment study, the osteoblasts labeled with 3H-leucine were incubated with the bio-derived bone materials in sterile microcentrifugable tubes for 15, 90 and 180 minutes, and 24 hours, respectively. The attached cells were collected and the radioactivity was measured with liquid scintillation spectrometry. In the proliferation study, the osteoblasts were cultured with the bio-derived bone materials for 24 hours and 3H-thymidine was added during the last 2 hours of the incubation. The attached cells were collected and the radioactivity was measured with liquid scintillation spectrometry. Osteoblasts were seeded on the bone graft materials for 60 or 120 minutes, 24 or 48 hours, and 3 or 7 days, then the co-culture was processed for scanning electron microscopy to observe the interaction of osteoblasts and the bio-derived bone materials.</p><p><b>RESULTS</b>Osteoblasts attached to the bio-derived bone materials in a time-dependent manner. There were significantly (P<0.05) more attached cells after 180 minutes than after 15 and 90 minutes of incubations (P<0.05). Osteoblasts were proliferated in a large amount on the surface and in the materials. Osteoblasts seeded onto 100 mg bio-derived bones resulted in significantly (P<0.05) more measurable proliferation than those seeded onto 10 mg bones. Osteoblasts appeared round as they attached to the materials, then flattened and spread over with time passing.</p><p><b>CONCLUSIONS</b>Bio-derived bones can provide a good environment for the attachment and proliferation of osteoblasts.</p>

Preparation of xenogenic corneal stroma and its cytoconsistency / 中国医学科学院学报

<p><b>OBJECTIVE</b>To explore the possibility that using the bovine corneal stroma to provide a suitable carrier on which the cells can grow for tissue engineering cornea.</p><p><b>METHODS</b>Nine fresh bovine corneas were selected. Each cornea was cut into 2 pieces, and exposed to 0.25% trypsinase for various lengths of time (20 minutes, 40 minutes, and 60 minutes) to get the stroma part with least cells and maintaining the collagen fibers arrangement. Samples obtained from each group were examined with scanning electron microscopy and HE staining. The left ones were freeze-dried and sterilized. The various concentrations of extraction were used to cultivate human fibroblasts, and a 3-(4,5-dimethylthiazol-2-yl)-2, (MTT)-based colorimetric assay was taken to evaluate the exhistance of 5-diphenyltetrazolium bromide cytotoxinic effects. Then the proper corneal stroma was used as a carrier to cultivated the rabbit corneal limbal cells which were planted on it in a concentration of 2 x 10(5)/cm2 in vitro. The cell-carrier samples were sent for scanning electron microscopy and HE staining.</p><p><b>RESULTS</b>The corneal stroma had the least cells in the group acted with typsin for 60 minutes, while the collagen fibers arrangement was not so orderly as before. The extractions showed no significant difference in cell culture, and no obviously harmful effect on the cell growth. The rabbit corneal limbal cells presented a stratified growth on the bovine corneal stroma.</p><p><b>CONCLUSION</b>The bovine corneal stroma without cells prepared using the typsin and lyophilization can be a suitable carrier for cell culture in vitro.</p>

Study on association between the expression of galectin- 3 and the peritoneal metastasis in gastric cancer / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To investigate the association between the expression of galectin- 3 protein and peritoneal metastasis in gastric cancer.</p><p><b>METHOD</b>The expressions of galectin- 3 was detected in matching- samples including primary gastric cancer lesions,lymph node metastases,peritoneal metastases and paratumor normal tissues by immunohistochemistry. All specimens were gained from 35 patients who had synchronous peritoneal metastasis from gastric cancer.</p><p><b>RESULTS</b>The over- expression of galectin- 3 was observed in 97% (34/35) of the gastric cancer lesions, the peritoneal metastases and the lymph node metastases,whereas in 14% (5/35) of paratumor normal tissues. There were significant differences in the expression of galectin- 3 between paratumor normal tissues and the gastric carcinoma lesions,peritoneal metastases and lymph node metastases (P< 0.05),but there were no significant differences among the gastric cancer lesions,the peritoneal metastases,and the lymph node metastases (P> 0.05).</p><p><b>CONCLUSION</b>The expression of galectin- 3 in gastric cancer lesions can be used as a biological marker of peritoneal metastasis from gastric cancer before operation and as a prognostic factor of gastric cancer.</p>

Observation of the morphological change during the adipogenesis differentiation of human bone marrow stromal cell / 中国康复理论与实践

@#ObjectiveTo study the variant differentiated phase of the human bone marrow stromal cell (MSC) during induced adipogenesis course by morphological observation.MethodsAfter proliferated in vitro, MSCs isolated from bone marrow of the female adults volunteers were cultured with the adipogenetic inducers for 6—30 days. Morphological changes of the cells were observed everyday under Olympus contrast phase microscopy, and some specimens were stained with Oil-red-O.ResultsMSC showed long spindle shape before inducing, and then changed gradually to oval or round shape and the figure enlarged simultaneously. There were specifically morphological changes with lipid droplet emergence under plasmalemma and confluence gradually to lipid drop during differentiated into the phase of immature and mature adipocyte.ConclusionIt is easy to detect the MSC differentiated into the phase of immature and mature adipocyte with the specific morphological change of lipid droplet, while in the phase of preadipocyte and adipoblast, there is no special morphological change, and it may need some specific markers to detect.