Objective To investigate the expression of basic fibroblast growth factor(bFGF) and proapoptotic gene Bax,Bcl-2 in amniochorion and their relationship with premature rupture of membranes(PROM).Methods Immunohistochemical stain was used to detect the expression of bFGF,Bax and Bcl-2 in amniochorion collected from 30 pregnant women with PROM and 30 normal pregnant women without PROM.Results The expression of bFGF and Bax were located in the plasma of all types of cells including trophoblast,epithelial,mesenchymal and fibroblast,but the expression of Bcl-2 was only observed in the chorionic syncytiotrophoblast;the positive average integral calculus oculus dehtor of bFGF in the PROM group was significantly lower than that of the normal group(0.051?0.012 vs 0.098?0.027,P0.05).Negative correlation was found between the bFGF and Bax expression in the PROM group(r=-0.616,P0.05). Conclusions The reduced expressions of bFGF and excessive apoptosis in the amnio-chorion play an important role in PROM.

Objective The gastrointestinal tract of neonate becomes colonized with microorganisms from environment and mother immediately after birth. Strong evidences suggested that early composition of the microflora in neonates play an important role for postnatal development of the immune system. This study was designed to investigate the effect of mode of delivery on the development of gut microflora and characteristics of the stool in breast-fed infants by molecular biology methods. Methods Sixty healthy breast-fed term infants were enrolled in the study(including vaginal delivery group and cesarean section delivery group,each group included 30 infants). Anthropometric measurements and stool study were done at 6-week,8-week,10-week,and 12-week. Fecal samples were subjected to quantitative fluorescence real-time polymerse chain reaction assays for the enumeration of lactobacili,bifidobacteria,and subtype of bifidobacteria(bifidobacterium longum and bifidobacterium breve). Results The numbers of lactobacilli and bifidobacterium longum species were less in cesarean delivery group than those in vaginal delivery group(P ＜ 0.05). Stool mean pH was lower in vaginal delivery group than that in cesarean section delivery group(P ＜ 0.05). There were no differences in stool characteristics and growth between two groups. Conclusions Cesarean section is associated with the decrease of bifidobacteria and lactobacilli in breast-fed term infants.

Objective To evaluate the influence of up-regulation of glucose regulated protein 78 (GRP 78)induced by 2-deoxyglucose(2DG)on fetal rat cerebral neuron apoptosis following intrauterine distress and the unification of endoplasmic reticulum and mitochondrium.Methods (1) Fetal rat intrauterine distress model was established and rats were divided into normal group(n=10),ischemiareperfusion(IR) group( n = 40) and treatment group ( n = 40, injection of 2DG into pregnant rats' abdomen after operation ). (2) Neuron apoptosis and the influence of 2DG on apoptosis was detected by terminal deoxynucleotidyltransferase-mediated dUTP-biotin nick end labeling (TUNEL) staining. The expression of GRP78, caspase-9,-12,and cytoron C protein were detected by western blot technique. Results (1) The number of TUNEL positive neuron in normal group was 4. 3±1. 8 /mm2. The expression of GRP78,caspase-9,-12, cytoron C in cytoplasm were 0.012±0.003, 0.004±0.003, 0.006±0.002, 0.012±0. 005, respectively. (2) The number of TUNEL positive neuron in the IR group were 43.6±11.4/mm2( reperfusion 3 h), 64. 4±9. 3/mm2 (repeffusion 6 h), 74. 2±12. 1/mm2 ( repeffusion 12 h), 97. 3±8. 9 /mm2 (reperfusion 24 h), respectively. They were significantly more than that in normal group(P ＜0. 05).The expression of GRP78 at corresponding times in IR group were 0. 092±0. 008 ( reperfusion 3 h), 0. 078±0.006 (reperfusion 6 h), 0.054±0.009 (reperfusion 12 h), 0.038±0.007 (reperfusion 24 h),respectively. The expression of cytoren C in cytoplasm at corresponding times in IR group were 0. 040±0. 006 (repeffusion 3 h), 0. 076±0. 009 (reperfusion 6 h), 0. 108±0. 005 (reperfusion 12 h), 0. 089±0. 008 (reperfusion 24 h), respectively. The expression of caspase-9 at corresponding times in IR group were 0. 042±0. 003 ( reperfusion 3 h), 0. 086±0. 007 ( reperfusion 6 h ), 0. 142±0. 006 ( reperfusion 12 h), 0. 112±0. 009 ( reperfusion 24 h), respectively. The expression of caspase-12 at corresponding times in IR group were 0. 076±0. 006 (reperfusion 3 h), 0. 113±0. 010 (reperfusion 6 h), 0. 125±0. 005 (reperfusion 12 h), 0. 057±0. 008 (reperfusion 24 h), respectively. They were significantly higher than that in normal group(P＜0. 05). (3) The number of TUNEL positive neuron in the treatment group were 19.4±10. 6/mm2 ( reperfusion 3 h), 26. 4±12. 3/mm2 ( repeffusion 6 h), 39. 3±13.3/mm2 ( reperfusion 12 h), 49. 3±13. 6/mm2 (reperfusion 24 h), respectively. They were significantly lower than that in IR group, but more than that in normal group(P＜0. 05). The expression of GRP78 at corresponding times in the treatment group were 0. 158±0.012 (repeffusion 3 h), 0. 175±0. 005 (reperfusion 6 h), 0. 125±0. 013 (reperfusion 12 h), 0. 079±0. 004 (reperfusion 24 h), respectively. They were significantly higher than that in IR group and normal group (P＜0. 05 ). The expression of cytoron C in cytoplasm at corresponding times in IR group were 0. 026±0. 002 (reperfusion 3 h), 0. 042±0. 008 (repeffusion 6 h),0. 062±0. 007 ( reperfusion 12 h), 0. 045±0. 004 ( reperfusion 24 h), respectively. The expression of caspase-9 at corresponding times in IR group were 0. 033±0. 002 ( reperfusion 3 h), 0. 063±0. 005(reperfusion 6 h), 0. 092±0. 005 (reperfusion 12 h), 0. 068±0. 008 (reperfusion 24 h), respectively.The expression of caspase-12 at corresponding times in IR group were 0. 061±0. 004 ( reperfusion 3 h),0. 068±0. 009 ( reperfusion 6 h), 0. 072±0. 007 ( reperfusion 12 h), 0. 054±0. 005 ( repedusion 24 h),respectively. They were significantly lower than that in IR group, but higher than that in normal group(P＜0. 05). Conclusions Fetal rat cerebral neuron apoptosis following intrauterine distress is associated with the action of endoplasmic reticulum and mitochondrium. Up-regulation of GRP78 induced by 2DG counteracts primary cellular damage caused by endoplasmic reticulum stress. 2DG plays a protective role for fetal rat cerebral neuron following intrauterine distress.

In this paper, the specific male sequeuce in paraffin section of various tissues with autolys is ofdifferent degrees was detected by polymerase chain reaction(PCR).The results shewed that PCR could be used for identifying sex in the tissues with low degrees ofautolysis; its positive rate was lower in high degrees with disappearance of nuclaic member and cell ou-tline,and often gave false negative results when the autolysis degree was high.

Objective To investigate the ex tr action technique for seperating the active components in the root of Salvia mi ltiorrhizae bunge by supercritical fluid, and to analyze the extracted product s by HPLC-MS n . Methods The extraction condition s were established as follows: 950ml?L -1ethanol as the first entrainer, t he pressure of 20.0 MPa, temperature at 45 ℃, and extracting time 1 h; then 100 mL?L -1 ethanol was selected as the second entrainer, pressur e was 30.0 MPa, temperature was 65 ℃, and extracting time was 3 h. Results Compared with traditional refluxing extraction and ultrasonic extraction, supercritical fluid extraction was better and more effect ive. Conclusion Supercritical extraction is simple, highly selec tive and efficient in extracting the active components in Salvia miltiorrhizae bunge.

As early as the 20 th century, it has been observed that radiotherapy (RT), as a local therapy, can activate the adaptive immune system, resulting in spontaneous regression of tumors out of the radiation field, which is known as "abscopal effect". Although the occurrence of abscopal effect is still rare, with the gradual increase in the application of immunotherapy, more and more clinical cases of abscopal effect have been reported. Increasing attention has been paid to the therapeutic potential of RT in inducing systemic anti-tumor response. Especially, the combination of RT and immunotherapy enhances the research value of abscopal effect. However, its mechanism has not been fully elucidated, and the optimal timing, dose and fractionation of RT are also under study. How to classify the beneficiary groups is also a key issue. In this article, the history of abscopal effect, and the role of RT and immunotherapy in this phenomenon were briefly introduced, and the existing controversies in clinical application were illustrated, aiming to clarify the direction of current research and development and open a new chapter for tumor treatment in a short period of time.

Objective:This study aimed to investigate the possible mechanism of 32P colloid induced apoptosis of craniopharyngi-oma (CP) cells in vitro and the relationship between dose effect and time effect. Methods:This study established a primary cell culture of CP limited subculture cell line. Methyl thiazolyl tetrazolium (MTT) assay was performed to plot the cell survival curve after the CP cells were treated with 32P colloid at different concentrations and time. Apoptotic rate was detected by flow cytometry(FCM). Apoptosis related DNA was investigated by TUNEL fluorescent staining. The morphological characteristics of apoptotic cells were determined by Hoechst33342 fluorescence staining. The ultrastructure of apoptotic cells was investigated by transmission electron microscopy (TEM). Results:Hoechst33342 fluorescence staining, TUNEL fluorescence staining, and TEM revealed that 32P colloid induced the apoptosis of CP cells. 32P colloid reduced the survival rate and increased the apoptotic rate of CP cells as concentration (0 MBq/mL to 14.80 MBq/mL) and time (1 d to 14 d) were increased. Conclusion: 32P colloid could effectively inhibit the growth of CP cells and induce apoptosis in vitro. High concentrations and prolonged time could induce a remarkable effect.

Objective To investigate the effect of butylphthalide (NBP) on H2S content and the expression of NR2B in the hippocampus of alcohol dependence rats. Methods A total of 84 SD male rats were randomly divided into 6 groups. Except for the normal group, other groups were subjected to alcohol solution with concentration of 6% ( V/V) for 28 d. Drug intervention began at the 14th day,and rats in the low,medium,high dose group were treated with NBP with a different concentration. Erden abstinence scoring was used to evaluate the rats withdrawal symptom. H2S content was measured in one side of hippocampus and CBS activity was tested in the other side of hippocampus. Hippocampus of 3 rats from each group was used to investigate NR2B mRNA level. Results Withdrawal symptom score ( 12.27 ± 1. 19),H2S content(30. 25 ±8.82), CBS activity (72. 44 ±7. 46) and NR2B mRNA expression( 19. 47 ±0. 86) in medium dose NBP group rats were lower than withdrawal symptom score(14.09 ±2.21) ,H2S content(44. 50 ±6. 65) , CBS activity(79. 06 ±4. 57) and NR2B mRNA expression (29. 13 ±1.39) in experimental control group (P<0.05). Withdrawal symptom score(12. 18 ±1.08) ,H2S content(33.00 ±5.38) ,CBS activity(67. 81 ±9. 37) and NR2B mRNA expression(23. 12 ± 1. 86) in high dose NBP group rats were lower than experimental control group (P < 0. 05). Conclusion NBP can reduce withdrawal symptoms of alcohol dependence rats,may be related to decreased expression of H2S/CBS system, and NR2B mRNA expression.

Objective To obtain colon cancer spheroid cells from human colon cancer cell lines cultured in serum-free medium (SFM),and investigate the proliferative and migratory properties of colon cancer spheroid cells.Methods Human colon cancer cell lines HCT116 and HT29 were cultured in SFM,and then the generation of spheroid cells was observed.The expression of stem cell surface marker CD133 was detected by flow cytometry,and the proliferative and migratory properties of colon cancer spheroid cells were detected by cell counting kit-8 and Transwell migration assay,respectively.All data were analyzed by using the t test.Results Spheroid cells were obtained from colon cancer cell lines HCT116 and HT29 in SFM.The ratios of spheroid cells with positive expression of CD133 generated by HCT116 and HT29 were 75.44％ ± 11.41％ and 76.22％ ± 14.23％,respectively.Compared with original colon cancer cells cultured in serum supplemented medium,the number of HCT116 and HT29 spheroid cells with positive expression of CD133 was significantly greater (t =11.43,9.17,P ＜ 0.05 ),and the proliferative and migratory abilities were much stronger also.Conclusion Colon cancer spheroid cells cultured in SFM have higher positive expression of CD133 and stronger proliferative and migratory abilities,and it can be utilized as a feasible model for further studies of colonic stem cells.

Objective To explore the status of mutations of k-ras gene in colorectal cancer (CRC) patients and to make theory preparation for the k-ras mutation detection in diagnosis laboratory. Methods The Genomic DNA was extracted, mutation analysis of k-ras was detected by PCR and bi-direction sequencing in the 56 specimens. Results Rate of k-ras mutation was 46.63 % (26/56) including 76.92 % (20/26) located at codon 12, and 23.08 %(6/26) located at codon 13, and no mutation was found at both codons simultaneously. G>A transition is the most common type of k-ras mutation,GGT>GAT (G12D) is the predominant mutation at codon 12 and GGOGAC (G13D) at codonl3. Chi-square analysis revealed the k-ras mutation was significantly correlated to the gender of the patients. Conclusion The k-ras mutation is mainly located at the codon 12, G>A transition is the most type of k-ras mutation in CRC. k-ras mutation seems to correlate with the gender of CRC patients.