A Multi-center, Randomized, Controlled and Open Clinical Trial of Heyan Kuntai Capsule () and Hormone Therapy in Perimenopausal Women / 中国结合医学杂志

<p><b>OBJECTIVE</b>To evaluate the efficacy and safety of Heyan Kuntai Capsule (, HYKT) and hormone therapy (HT) on perimenopausal syndromes (PMSs).</p><p><b>METHODS</b>From 2005 to 2008, 390 women with PMSs were recruited from 4 clinic centers. The inclusion criteria included ages 40 to 60 years, estradiol (E2) below 30 ng/L, and follicle stimulating hormone (FSH) above 40 IU/L, etc. The patients were randomly assigned to HYKT group or HT group by random number table method, administrated HYKT or conjugated estrogen with/without medroxyprogesterone acetate tablets for 12 months. During treatment, the patients were interviewed quarterly, Kupperman Menopausal Index (KMI) scores, hot flush scores, insomnia scores, Menopause-Specific Quality of Life (MENQOL) scores and adverse effects were used for evaluating drug efficacy and safety respectively. The last interview was made at the end of 12-month treatment RESULTS: After treatment, KMI scores of HYKT group and HT group were both significantly decreased compared with baseline (P <0.01) and there was no significant difference between groups (P >0.05), except that KMI of HYKT group was higher after 3-month treatment (P <0.05). After treatment, hot flush and insomnia scores were both improved significantly in two groups (P <0.01); and HT had a better performance than HYKT in improving hot flush (P <0.05). MENQOL were significantly improved in both groups after treatment (P <0.01); but there was no significant difference between two groups (P >0.05). The incidence of adverse event in the HYKT group was much lower than that in the HT group (P <0.01).</p><p><b>CONCLUSIONS</b>HYKT could effectively relieve PMSs and improve patients quality of life without severe adverse reactions. Although HYKT exerted curative effects more slowly than hormone, it possessed better safety profile than hormone.</p>

Adsorptive Granulocyte and Monocyte Apheresis in the Treatment of Ulcerative Colitis: The First Multicenter Study in China

BACKGROUND/AIMS: Patients with active ulcerative colitis (UC) have elevated levels of activated myeloid-derived leukocytes as a source of inflammatory cytokines. The selective depletion of these leukocytes by adsorptive granulocyte/monocyte apheresis (GMA) with an Adacolumn should alleviate inflammation, promote remission and enhance drug efficacy. However, studies have reported contrasting efficacy outcomes based on patients’ baseline demographic variables. This study was undertaken to understand the demographic features of GMA responders and nonresponders. METHODS: This was a multicenter study in China involving four institutions and 34 patients with active UC. Baseline conventional medications were continued without changing the dosage. The treatment efficacy was evaluated based on the endoscopic activity index and the Mayo score. RESULTS: Thirty of the 34 patients completed all 10 GMA treatment sessions. The overall efficacy rate was 70.59%. The receiver operating characteristic analysis showed that the area under the curve was approximately 0.766 for a Mayo score of ≤5.5 with 0.273 specificity and 0.857 sensitivity (Youden index, 0.584) for GMA responders. No GMA-related serious adverse events were observed. CONCLUSIONS: The overall efficacy of GMA in patients with active UC who were taking first-line medications or were corticosteroid refractory was encouraging. Additionally, GMA was well tolerated and had a good safety profile.

Establishment and validation of human cancer cell lines with stable Cas9 expression / 中华病理学杂志

Objective@#To establish human cancer cell strains with stable Cas9 expression, and to validate the gene editing activity of Cas9 for simple gene editing in future study.@*Methods@#Fifteen cancer cell lines of different tissue origins were infected with pLv-EF1α-Cas9-Flag-Neo or pLv-EF1α-Cas9-Flag-Puro by lentivirus and clone selection was employed to screen Cas9 stably expressed cancer cell lines. Afterward designed guide RNA vectors targeting TSC22 gene were transiently transfected into 3 of cell lines, and subsequently the gene editing activity of Cas9 was evaluated by genomic PCR, sequencing and Western blot.@*Results@#Sixty-nine human cancer cell strains with stable Cas9 expression from different cancers were established, and by transient transfection with designed guide RNA, long fragment deletion was detected in TSC22 gene.@*Conclusions@#Sixty-nine human cancer cell strains are successfully established with stable expression of Cas9 protein and gene editing activity. These cell strains may be employed in large-scale drug screening, screening of new drug targets and gene function investigation.

Effect of Chinese herbal compound Naofucong () on the inflammatory process induced by high glucose in BV-2 cells / 中国结合医学杂志

<p><b>OBJECTIVE</b>To determine the effect of medicated serum of Chinese herbal compound Naofucong (, NFC) on the microglia BV-2 cells viability and the transcription and expression of interleukin-6 (IL-6) and tumor necrosis factor α (TNF-α) in microglia BV-2 cells to further explore the mechanisms underlying the protective effect of NFC on inflammatory process induced by high glucose.</p><p><b>METHODS</b>The microglia BV-2 cells incubated in vitro were divided into different groups: the control group (25 mmol/L glucose), the model group (75 mmol/L glucose), high glucose media containing different dose medicated serum of NFC. After being cultured for 24 h, changes in IL-6 and TNF-α were measured by quantitative real-time polymerase chain reaction and enzyme-linked immunosorbent assay. The expression of surface marker CD11b of activated microglia was measured by confocal laser scanning microscope and Western blot. Nuclear factor-κB (NF-κB) p-p65 expression was analyzed by Western blot.</p><p><b>RESULTS</b>The model group obviously increased the expression of microglial surface marker CD11b and NF-κB p-p65 (all P<0.01), induced a signifificant up-regulation of release and the mRNA expression of IL-6 and TNF-α (P<0.01 or P<0.05). The medicated serum of NFC could obviously down-regulate the transcription and expression of surface marker CD11 b and NF-κB p-p65 (all P<0.01), and inhibit the mRNA and protein expression (P<0.01 or P<0.05) of inflflammatory cytokines, such as IL-6 and TNF-α, in microglia BV-2 cells cultured with high glucose for 24 h.</p><p><b>CONCLUSIONS</b>The inhibition of microglial activation and IL-6 and TNF-α expression induced by high glucose may at least partly explain NFC therapeutic effects on diabetes-associated cognitive decline diseases. Its underlying mechanism could probably be related to the inhibition of NFC on NF-κB phosphorylation.</p>

Jinmaitong alleviates the diabetic peripheral neuropathy by inducing autophagy / 中国结合医学杂志

<p><b>OBJECTIVE</b>To observe the deregulation of autophagy in diabetic peripheral neuropathy (DPN) and investigate whether Jinmaitong ( JMT) alleviates DPN by inducing autophagy.</p><p><b>METHODS</b>DPN models were established by streptozotocin-induced diabetic rats and Schwann cells (SCs) cultured in high glucose medium. The pathological morphology was observed by the improved Bielschowsky's nerve fiber axonal staining and the Luxol fast blue-neutral red myelin staining. The ultrastructure was observed by the transmission electron microscopy. Beclin1 level was detected by immunohistochemistry and Western blot. The proliferation of cultured SCs was detected by methylthiazolyldiphenyl-tetrazolium bromide.</p><p><b>RESULTS</b>Diabetic peripheral nerve tissues demonstrated pathological morphology and reduced autophagic structure, accompanied with down-regulation of Beclin1. JMT apparently alleviated the pathological morphology change and increased the autophagy [in vivo, Beclin1 integral optical density (IOD) value of the control group 86.6±17.7, DM 43.9±8.8, JMT 73.3 ±17.8, P<0.01 or P<0.05, in vitro Beclin1 IOD value of the glucose group 0.47±0.25 vs the control group 0.88±0.29, P<0.05]. Consequently, inhibition of autophagy by 3-methyladenine resulted in a time- and concentration-dependent decrease of the proliferation of SCs (P<0.05, P<0.01).</p><p><b>CONCLUSIONS</b>Down-regulation of autophagy in SCs might contribute to the pathogenesis of DPN. JMT alleviates diabetic peripheral nerve injury at least in part by inducing autophagy.</p>

Observation on the growth and metastasis of cross-strain transplanted tumors in different mouse strains / 中华肿瘤杂志

<p><b>OBJECTIVE</b>Mouse tumors were subcutaneously transplanted into different mouse strains and their growth and metastatic properties were checked, to explore the possibility of establishing animal tumor models in different mouse strains other than their normal host strains.</p><p><b>METHODS</b>Seven mouse tumor cell lines: H22, S180, U14, FC, Ca761, SMG-A and DCS were transplanted into C57BL/6J, ICR or KM mice, and their tumorigenicity, growth and metastasis were recorded and analyzed.</p><p><b>RESULTS</b>The tumor formation rate of H22 cells in both the C57BL/6J and ICR mice was 100%, but the growth of H22 tumors was significantly faster in the C57BL/6J (2.8 ± 0.4)g than in the ICR mice (1.5 ± 0.5)g at the 17th day after transplantation (P<0.001). The S180 tumors grew stably in C57BL/6J mice and the tumor formation rate was 100%. The U14 inoculated into C57BL/6J and KM mice showed both lymphatic and lung metastasis and formed significantly larger tumors in KM mice [(12.6 ± 3.4)g] than that in the C57BL/6J mice [(10.2 ± 2.2)g] on the 32rd day after transplantation (P = 0.002). Transplantation of FC, Ca761, and SMG-A did not form tumors or the tumors were completely regressed later in C57BL/6J mice. DCS cells formed tumors in C57BL/6J mice, but some of the tumors regressed. The retained tumors were passaged in C57BL/6J mice, and the substrain DCS-C57 cells was established which showed stable growth and had a 100% tumor formation rate and 100% lung metastasis rate in C57BL/6J mice.</p><p><b>CONCLUSIONS</b>Cross-strain transplanted tumors can be successfully established by inoculation of poorly differentiated and highly malignant tumor cells into different mouse strains. Some highly immunogenic tumor cells may form tumor, however, the tumors are regressed later, and can not establish cross-strain transplanted tumors in other mouse strains. Stable transplanted tumor models can be obtained from the partially regressed tumors after continuous passages in vivo.</p>

Effect of forced E-cadherin expression on adhesion and proliferation of human breast carcinoma cells / 中华病理学杂志

<p><b>OBJECTIVE</b>To investigate the role that E-cadherin (E-cad) plays on cell adhesion and proliferation of human breast carcinoma.</p><p><b>METHODS</b>E-cad expression vector was transfected into an E-cad-negative human breast carcinoma MDA-MB-231 cells. G418 was used to screen positive clones. E-cad, β-catenin (β-cat) and cyclin D1 expressions of these clones were confirmed by Western blot. Their cell-cell and cell-matrix adhesion abilities were detected. E-cad/β-catenin interaction was confirmed by immunoprecipitation. Cell proliferation was evaluated by MTT. Cell apoptosis was analyzed by flow cytometry. Direct two-step immunocytochemistry was used to detect the localization of β-cat.</p><p><b>RESULT</b>E-cad(+) cell strains Ecad-231-7 and Ecad-231-9 were established. When cultured in ultra-low-binding dishes Ecad-231 cells grow in suspension while Ecad-231-7 and Ecad-231-9 cells grow in large clamps. When co-cultured with HCT116 cells, the average adhesion rates at 30 min are 39.0%, 60.0% and 59.5% for MDA-MB-231, Ecad-231-7 and Ecad-231-9 respectively. The average detachment rates by EDTA for 5 min are 37.4%, 4.2% and 7.4% respectively. So E-cad expression enhanced hemotypic and heterotypic cell-cell adhesion and cell-matrix adhesion. Forced exogenously expressed E-cad could combine with endogenous β-cat, whereas down stream cyclin D1 expression was significantly decreased, as evidenced by Western blot. The rates of cell apoptosis of MDA-MB-231, Ecad-231-7 and Ecad-231-9 were 1.8%, 2.0% and 2.1%. Expression of E-cad had no obvious effect on the apoptosis of tumor cells with regular culture. β-cat increased in the cytoplasma.</p><p><b>CONCLUSIONS</b>Two monoclonal tumor cell strains (Ecad-231-7 and Ecad-231-9) stably expressing E-cad were successfully established. E-cad could enhance adhesion and inhibit proliferation of human breast carcinoma cells through a pathway involving β-cat and cyclin D1.</p>

Expression and function of VAP-33 in murine dendritic cell sarcoma / 中华病理学杂志

<p><b>OBJECTIVE</b>To elucidate the expression and function of VAP-33 gene in dendritic cell sarcoma (DCS) cell line.</p><p><b>METHODS</b>The expression of VAP-33 in DCS cells was investigated by mass spectrum with immunoprecipitation membrane protein. DCS cells were treated with antigens in different dosages (150, 850, and 1500 microl) for 24, 48 and 72 h respectively. Cell morphology and phagocytosis activity of DCS cells were measured. Indirect immunofluorescence, confocal microscopy and Western blotting were used to study the distribution and expression changes of VAP-33. Moreover, DCS cells were treated with 0.5 mol/L insulin for 20 min first and followed by Western blotting to detect changes of VAP-33 and glucose transfer protein 4 (GLUT-4) in the total cellular protein, cytoplasmic protein and membrane protein. Confocal microscopy was used to document the expression and distribution changes of VAP-33 and GLUT-4 in DCS cells.</p><p><b>RESULTS</b>VAP-33 expression was obtained at the cell membrane and in the cytoplasm of DCS cells. Upon antigen stimulation, DCS cells showed more active phagocytosis and morphologically became more elongated with branched protrusions. The expression of VAP-33 was decreased by the antigen stimulation. Upon the insulin stimulation, the expression of VAP-33 and GLUT-4 were increased and co-localized.</p><p><b>CONCLUSIONS</b>VAP-33 expression in DCS originated from the dendritic cells (DC) seemed relating to the vesicle transportation during antigen processing in DC. Additionally, VAP-33 and GLUT-4 also take part in the glucose transportation in the cells.</p>

Establishment of green-fluorescent protein expressing tumor metastasis models / 中华病理学杂志

<p><b>OBJECTIVE</b>To establish a green-fluorescent protein (GFP) labeled tumor metastasis model and to evaluate its biological characteristics.</p><p><b>METHODS</b>Human gastric carcinoma cell MGC-803 and murine cervical carcinoma cell U14 were transfected with the plasmid pEGFP-N1 and the efficiency of transfection was assessed 24 h later. Limited dilution was employed to screen and establish monoclonal cell strains, MGC-803-GFP and U14-GFP. The two fluorescent tumor cell stains were transplanted into BALB/c-nu mice and C57BL/6J mice respectively. The latency period of tumor mass appearance and the growth curve in vivo were documented. The tumor growth and metastasis were evaluated in vivo by the Viviperception Fluorescence Imagining System (VFIS). Expressions of CD44 and E-cadherin in tumor tissue were monitored by immunohistochemistry.</p><p><b>RESULTS</b>The efficiency of pEGFP-N1 transfection of MGC-803 cells and U14 cells were 30% and 60%, respectively. Monoclonal GFP(+) cell strains-MGC-803-GFP and U14-GFP were established. The latency periods of tumor formation of MGC-803-GFP and U14-GFP were 3-5 days and 2-4 days, respectively. Their tumorigenicity rates were 100% in both. The tumor growth of MGC-803-GFP was well defined by the VFIS. Only one mouse was shown to harbor lymphatic metastasis by VFIS, 60 days after transplantation. The metastasis process of U14-GFP was depicted through VFIS on 27, 37 and 52 days post-transplantation. The incidence of pulmonary metastasis and lymphatic metastasis of U14-GFP was 67% and 100% respectively when the tumor volume was >or=5 cm3. CD44 was positive and E-cadherin was negative in both tumors by immunohistochemistry.</p><p><b>CONCLUSIONS</b>Successfully established two monoclonal tumor cell strains stably expressing GFP: MGC-803-GFP and U14-GFP. Transplantation of these cells into mice can establish tumor metastasis models which could be used for future visualized tumor research in vivo.</p>

Inhibition effect of rhEPO on rhIL6-induced hepcidin mRNA expression in HepG2 cell lines and human primary hepatocytes / 中国实验血液学杂志

This study was aimed to investigate the effect of rhIL-6 and rhEPO on hepcidin mRNA expression in HepG2 cells and human primary hepatocytes, and mechanism of rhEPO in treatment of anemia of chronic disease (ACD). The HepG2 cells and human primary hepatocytes were cultured with medium containing different concentrations of rhIL-6 and rhEPO for a certain time, then mRNA was isolated and its RT-PCR was performed, the bands were photographed and analyzed by UVI band, the hepcidin and G3PDH mRNA ratio were semi-quantitatively analyzed. The expression levels of hepcidin in GepG2 cells and human primary hepatocytes at different conditions were compared. The results showed that the hepcidin mRNA expression in HepG2 cells and human primary hepatocytes could be enhanced by rhIL-6, the rhEPO could inhibit rhIL6-induced hepcidin mRAN expression. The rhEPO alone basically did not influence hepcidin mRNA expression in HepG2 cells. It is concluded that Hepcidin mRNA expression in HepG2 cells and human primary hepatocytes can be elevated by rhIL-6 with concentration- and time-dependent manner in certain range. rhEPO can inhibit this effect of rhIL-6.

Effect of inhibitor of differentiation-1 on murine dendritic cell sarcoma cells / 中华病理学杂志

<p><b>OBJECTIVE</b>To investigate the effect of down-expression of inhibitor of differentiation-1 (Id-1) on the differentiation of dendritic cell sarcoma (DCS) cells in vitro.</p><p><b>METHODS</b>Down-regulation of the expression of Id-1 in DCS cells was performed by RNAi, and confirmed by protein and mRNA quantitative analyses. Cellular differentiation and biological behavior including malignant phenotypes of the cells were evaluated. All experiments included negative (no treatment group and no-target siRNA) and positive (induction-differentiation drug sodium butyrate) controls.</p><p><b>RESULTS</b>When the expression of Id-1 was down regulated, the DCS cells showed more mature morphology including cell enlargement, longer cellular extensions, more branches, and decreased nuclear/plasma ratio. Differentiation marker expression (Id-2 and CD86) was also increased. RNAi treated cells at 24 and 48 hours, showed increase percentage of cells at G0/G1 phase and less cells at S phase (P < 0.01). Importantly, the abilities of cell proliferation, colony formation and invasiveness were significantly decreased (P < 0.01), as evidenced by MTT, colony formation and transwell assays respectively.</p><p><b>CONCLUSION</b>RNAi inhibition of Id-1 protein can induce differentiation of malignant solid tumor cells along with reversion of their malignant phenotype.</p>

Effect of Jinmaitong serum on the proliferation of rat Schwann cells cultured in high glucose medium / 中国结合医学杂志

<p><b>OBJECTIVE</b>To investigate the effect of Jinmaitong (JMT) serum on the proliferation of rat Schwann cells (SCs) primarily cultured in high glucose medium.</p><p><b>METHOD</b>SCs were primarily cultured in Dulbecco's minmum essential medium (DMEM control), 50 mmol/L glucose medium (50 mmol/L Glu), 75 mmol/L glucose medium (75 mmol/L Glu), as well as 50 mmol/L glucose medium, with different concentrations of JMT serum (undiluted, 1:2 diluted and 1:8 diluted) and Neurotropin (Ntp), respectively. The proliferation of SCs under different conditions was detected by MTT.</p><p><b>RESULT</b>SCs grew exuberantly in DMEM within 24-72 h, but slowed down at 96 h. The proliferation of SCs was inhibited in 50 mmol/L Glu and 75 mmol/L Glu after cultures of 48, 72 and 96 h, which showed that both were significantly different compared to the control group (P<0.01). The inhibition was more significant in 75 mmol/L Glu than in 50 mmol/L Glu (P<0.05). Spearman's rho analysis revealed that the proliferation of SCs had a negative correlation with the concentration of glucose (r=-0.471, P<0.01). Excluding the time factor, partial correlation showed similar results (r=-0.679, P<0.01). After 48 h, the proliferation of SCs increased significantly in JMT1:2 and Ntp compared with 50 mmol/L Glu (control 0.437+or-0.019, 50 mmol/L Glu 0.367+or-0.035, JMT1:2 0.426+or-0.024, Ntp 0.422+or-0.013; P<0.01), and there were no statistically significant differences among the JMT groups, the Ntp group and the control group (P>0.05).</p><p><b>CONCLUSIONS</b>The proliferation of SCs was inhibited in high glucose medium, and the inhibition was reduced by different concentrations of JMT serum, especially at JMT1:2.</p>

Establishment of animal model of breast cancer dormancy / 中华病理学杂志

<p><b>OBJECTIVE</b>Tumor dormancy has been defined clinically as a condition in which tumor cells are present but do not grow for a long period of time. Breast cancer is noted for its long periods of tumor dormancy and metastases can occur many years after treatment.</p><p><b>METHOD</b>Simulating the characteristics of breast cancer patients after treatment, we established the animal model of breast cancer dormancy by inoculating 500 Ca761-03 cells into the limb muscle of 615 mice and then selecting animals with tumor dormancy 2 months post inoculation (corresponding to 5 years for humans).</p><p><b>RESULTS</b>Two months after inoculation of Ca761-03 cells into the muscle of 615 mice, tumor occurred in 30% of the mice. The remaining 70% of mice did not show tumor growth. After repeated traumatic stimulation, 90% of the mice developed tumors after 5 months, therefore representing tumor dormancy.</p><p><b>CONCLUSIONS</b>These results demonstrate that breast cancer cells can remain in a dormant state for long periods of time in vivo. Trauma can stimulate the dormant tumor cells to proliferate again, and causes tumor relapse. This murine model system promises a sound animal model for the study of solid tumor dormancy.</p>

Effect of combination of eicosapentaenoic acid and epirubicin on human gastric carcinoma cell strain MGC-803 / 中国医学科学院学报

<p><b>OBJECTIVE</b>To evaluate the effect of combination of eicosapentaenoic acid (EPA) and The effects of EPA and epirubicin (EPI) on the human gastric carcinoma cell MGC-803 in vitro.</p><p><b>METHODS</b>EPI were measured by MTT assay , and the interaction between these two agents was evaluated by the isobologram technique of Berenbaum. Morphous of cell was observed by phase-contrast and electron microscope. Flow cytometry was used for cell cycle analysis.</p><p><b>RESULTS</b>EPA significantly inhibited the growth of MGC-803 cells in a dose- and time-dependent way (P < 0.01). Numerous abnormal particles were found around the nucleus of MGC-803 cells under phase-contrast microscope, and also many electron-dense material in cytoplasm were found under electron microscope. EPA significantly stimulated the growth of human embryonal pulmonary fibroblast (HPF) dose-dependently (P < 0.01). A strong synergism was found between EPA and EPI in MGC-803 cells. EPA induced G0/G1-phase arrest but without statistical significance (P > 0.05), and EPI significantly induced S-phase arrest (P < 0.05) in MGC-803 cells.</p><p><b>CONCLUSIONS</b>EPA can inhibit cell growth in gastric carcinoma cells but not in normal cells. EPA and EPI have synergetic effect in the inhibition of gastric carcinoma cells. Compared with EPI monotherapy, the combination of EPI and EPA can reduce the dosage of EPI.</p>

Effects of human alpha-mannosidase Man2c1 transgene on growth and metastasis of transplanted tumor in mice / 中国医学科学院学报

<p><b>OBJECTIVE</b>To study the effect of human alpha-mannosidase Man2c1 transgene on tumor growth and metastasis in mice.</p><p><b>METHODS</b>Hepatoma cell H22 or squamous epithelial carcinoma cell S180 was subcutaneously inoculated into the right armpit of mice (wild type mice and 28#, 35#, and 54# transgenic mice). Tumor size was measured every week. Mice were sacrificed on day 9 or 10 and then the tumors were exercised and weighted. Tumors and lungs were fixed in formaldehyde and sectioned. The sections were stained with hematoxylin/eosin and examined under microscope. The red blood cells in spleen were destroyed by Tris-NH4Cl. Natural killer (NK) cell activity was detected with Yac-1 cell as target.</p><p><b>RESULTS</b>H22 and S180 tumors grew faster in all the three transgenic mice (28#, 35#, and 54#) than in wild type mice. The average size and weight of tumors between the transgenic mice and wild type mice were significantly different (P<0.05). Most tumors in the transgenic mice invaded the surrounding tissues. In contrast, nearly all the tumors in wild type mice were capsulized. Three of 10 28# transgenic mice, 5 of 10 35# transgenic mice, 3 of 10 54# transgenic mice, and 1 of 10 wild type mice showed lung metastasis of H22 tumor. Two of 6 28# transgenic mice, 3 of 6 35# transgenic mice, 1 of 6 54# transgenic mice, and 0 of 6 wild type mice showed lung metastasis of S180 tumor. No difference of NK activity in spleen cells was observed between the transgenic mice and wild type mice.</p><p><b>CONCLUSIONS</b>hMan2c1 transgene promotes growth, invasion, and metastasis of transplanted H22 and S180 tumors in mice. hMan2cl transgene does not affect NK activity in splenocytes.</p>

Species identification of animal cells by polymerase chain reaction / 基础医学与临床

Objective To identify the species of cultured cells and to detect inter-species cross contamination by polymerase chain reaction (PCR).Methods From references and NCBI database,we outsourced 32 pairs of species-specific primers targeted to genomic DNA of ten common species of cultured cells.Then we screened for optional primers with high specificity and high sensitivity.PCR amplification with these primers,the genomic DNAs isolated from the tested cell line and agarose gel electrophoresis of the PCR products were done.Mixed DNA of 10 species was used as positive-control template,and water as negative-control template.Results Ten pairs of species-specific and highly sensitive primers were selected.By PCR amplification with these primers and agarose gel electrophoresis,we may easily identify the origin of cell lines and find whether the tested cell lines are contaminated by cells of other species.Conclusion This PCR assay provides a simple,rapid,sensitive,and cost-effective method to identify cell species and detect interspecies cross-contamination.

Culture and characterization of human fetal hepatocytes / 基础医学与临床

Objective Determining the characteristics of human fetal hepatocytes in vitro. Methods Isolating and culturing human fetal hepatocytes by stepwise trypsinization of liver fragment in vitro; collecting the culture medium to determine the secretion of AFP, ALB and the functional enzymes (including ALT, AST, GGT, ALP and LDH) of different generations in culture; determining the expression of cytochrome C by immunohistochemistry; testing the effects of sodium byturate on human fetal hepatocytes. Results Human fetal hepatocytes were polygonal epithelial cells in DMEM medium. They could be maintained for 5.5 months (about 30 passages) in vitro. They secreted ALB and functional enzymes all over their cultivation. Conclusion Human fetal hepatocytes can be maintained keeping function in vitro for several months.

Establishment and characterization of a rabbit tumor cell line VX2 / 中华病理学杂志

<p><b>OBJECTIVE</b>To establish a rabbit tumor cell line and to characterize its biological parameters.</p><p><b>METHODS</b>VX2 tumor tissue was used for the primary culture in vitro. After 40 passages, the cell morphology, CK expression (immunohistochemical staining), cell cycle, karyotype and tumorigenecity in rabbits and nude mice were investigated.</p><p><b>RESULTS</b>The newly established cell line VX2 was maintained in continuous culture for over 70 passages in 10 months. Morphologically, VX2 cells were polygonal to short spindled. Tonal fibril and tight junction were found under the electron microscope. CK was positive. The cell cycle analysis showed 69.3% in G1 phase, 5.6% in G2 phase and 25.1% in S phase. The population doubling time was 34.5 hours. The chromosomal analysis showed a hypotriploidy with a median chromosome number of 58 approximately 62. The tumorigenecity in rabbits and nude mice were both 100%.</p><p><b>CONCLUSION</b>The established VX2 cell line derived from rabbit squamous carcinoma could serve as a model system for experimental oncology in the rabbit.</p>

Influence of tumor cells on endothelial cells in vitro / 中华病理学杂志

<p><b>OBJECTIVE</b>To observe in vitro changes of endothelial cells after confrontation with tumor cells.</p><p><b>METHODS</b>Dynobeads were used to isolate the endothelial cells from the rat lung. Mouse dendritic cell sarcoma cells (DCS), human gastric carcinoma cells (BGC-823) and mouse lung adenocarcinoma cells (LA795) were added to the endothelial cells when the latter was at the confluence phase. Phase contrast microscope, scanning electro-microscope, immunohistochemistry, transwell and fluorescence dye transfer were used to detect morphological and functional changes of the endothelial cells.</p><p><b>RESULTS</b>Endothelial cells may look like cobble stones or long spindle shaped. Direct contact of tumor cells with endothelial cells induced round vascular-like space formation between confluent endothelial cells. Tumor cells were often found at the newly appeared spaces. Tumor cell conditioned medium could support the growth and promote the locomotion of endothelial cells through transwell. It was observed that luciffer yellow was directly transported from tumor cells to endothelial cells.</p><p><b>CONCLUSIONS</b>Tumor cells can directly induce morphological and functional changes in endothelial cells. Direct intercellular communication between tumor cells and endothelial cells is present.</p>