1.Expression of basic fibroblast growth factor(bFGF),Bax and Bcl-2 in amniochorion of premature rupture of membranes
Zongjie LU ; Li JIAN ; Hongbo QI
Chinese Journal of Perinatal Medicine 2000;0(04):-
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.
2.Up-regulation of glucose regulated protein 78 induced by 2-deoxyglnoose plays a protective role for fetal rat cerebral neuron following intrauterine distress
Hua ZHANG ; Min LU ; Hongbo QI ; Jianhua ZHANG
Chinese Journal of Obstetrics and Gynecology 2008;43(5):356-360
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.
3.SEX IDENTIFICATION OF AUTOLYSIS TISSUES BY POLYMERASE CHAIN REACTION
Yingqiang LU ; Zhaoyan XUAN ; Ming ZHANG ; Hongbo WANG
Chinese Journal of Forensic Medicine 1987;0(03):-
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.
4.Active components in the extracts of Radix Salvia miltiorrhizae by supercritical carbon dioxide fluid
Xia LI ; Yuhai TANG ; Xinfeng ZHAO ; Hongbo LU ; Xiaohui ZHENG
Journal of Xi'an Jiaotong University(Medical Sciences) 2003;0(06):-
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.
5.Impact of mode of delivery on gut microflora in breast-fed infants
Hongbo ZHANG ; Wei LU ; Juan HUANG ; Yexuan TAO ; Wei CAI
Journal of Clinical Pediatrics 2010;(3):251-254
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.
6.The roles and clinical controversies of radiotherapy and immunotherapy in abscopal effect
Lu ZHANG ; Hongbo LIU ; Xiao DING ; Haijun LU
Chinese Journal of Radiation Oncology 2023;32(5):470-475
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.
7.32P colloid induced apoptosis of craniopharyngioma cells in vitro
Hongbo CHANG ; Ming GAO ; Shuwei WANG ; Siyuan ZHAO ; Wangsheng LU ; Xin YU ; Zengmin TIAN ; Jianning ZHANG
Chinese Journal of Clinical Oncology 2014;(10):624-628
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.
8.Safety and validity of a new-type biological dura patch
Hongbo CHANG ; Tengfei PAN ; Wangsheng LU ; Peng WANG ; Kai CUI ; Jianning ZHANG
Chinese Journal of Tissue Engineering Research 2014;(25):3947-3952
BACKGROUND:Dural repair materials in current application mainly include autologous tissue repair material, alograft material, heterogeneous biological material and synthetic material, most of which are imported products with expensive price. OBJECTIVE: To evaluate safety and efficacy of a new biological type dura mater patch made in China based on animal experiments. METHODS:Bilateral dura mater defect models were established in 24 healthy domestic dogs: on the left side of the implant model, a new type biological dura patch was transplanted as experimental group; on the right side, another brand artificial dura patch that was on sale was transplanted as control group. After 1, 3, 6 and 12 months of implantation, we compared degradation, angiogenesis, growth and surrounding tissue reaction of dural substitutes of the experimental group and control group by hematoxylin-eosin staining, detected residual dose of epoxy-cross-linked agent in dogs’ blood and cerebrospinal fluid by fluorescence spectrophotometry. RESULTS AND CONCLUSION: During 1-12 months of implantation, al dogs grew wel and no infection or motor disorder was observed. Pathological examination showed that dura substitutes of the experimental group and control group had good biocompatibility, no or slightly inflammatory response. After 6 months of implantation, the surface of the new biological dural substitute (experimental group) was degraded and became a transit-state biomaterial with surrounding tissue, but the control group materials showed no degradation. After 12 months of implantation, the dura patch in the experimental group degraded nearly 50%, which appeared with neovascularization; while, the dura patch in the control group degraded 30%, and neovascularization was observed in only a smal amount of samples. Epoxy compounds of cross-linked agent were not detected in dogs’ blood and cerebrospinal fluid after 1, 3, 7 and 14 postoperative days. These findings show that this new type of biological dural substitute is a safe and effective dural repair material.
9.The effects of proanthocyanidins on antidepressant-like behaviors and the adrenal gland in chronic stress rats
Guixiang CHEN ; Jie YU ; Lu ZHANG ; Lujing WANG ; Huiying CAO ; Yijia YAN ; Hongbo SHAO ; Jianchun PAN
Chinese Journal of Behavioral Medicine and Brain Science 2010;19(7):611-613
Objective To investigate the effects of proanthocyanidins on depressant-like behaviors and the structure of adrenal gland in chronic unpredictable mild stress (CUMS) rats. Methods Rats were randomly divided into 6 groups:control group, stressed group (CUMS + vehicle), three treatment groups (CUMS + proanthocyanidins 25,50,100 mg·kg-1,respectively) ,and imipramine group (CUMS + imipramine 10 mg·kg-1). Used the CUMS model in rats to investigate the effects of chronic oral administration (21 days) of proanthocyanidins and imipramine (ip) on the open-field;and forced swimming and sucrose consumption tests and the ratio of adrenal gland/body weight,and its thickness were examined by HE stain. Results Compared with control group, rats subjected to CUMS exhibited increased ratio of adrenal gland /body weight ( P < 0. 01), less sucrose consumption( P<0.01) and inhibited in the open-field test( P<0.01) as well as more despair time in the forced swimming test( P<0.01). While compared with stressed group,treatments with proanthocyanidins (25,50,100 mg·kg-1, po ,21 days) could significantly improve the activities in open-field test ((39.6±3.4) vs (49±4.5), (52.6±3.7),(54.1±1.8) ;all P<0.01) and sucrose consumption( (5.8±2.5)ml vs (8.1±3.3)ml,(8.5±4.1) ml, (9.2±2.6) ml; P < 0.05, P < 0.05, P < 0.01 respectively); Meanwhile, it could reduce the duration time in forced swimming test significantly( (103.5±10.2)s vs (83.7±8.8)s,(75.8±5.9)s,(67.2±6.5)s; all P<0.01) as well as thickness of the adrenal gland(P<0.05, P<0.01).Conclusions This study suggests that the proanthocyanidins (25,50,100 mg·kg-1) has an antidepressant-like effects in CUMS rats. The antidepressant actions of proanthocyanidins, in some degrees, may be related with the regulation of the adrenal gland's structure.
10.The effect of buyanghuanwu decoctinn on expression of immunoreactive protein and mRNA of NMDA receptor 2B subunit in rats hippocampal with vascular dementia
Jun LI ; Weijuan GAO ; Guobing LU ; Jinglong TANG ; Hongbo ZHANG ; Tao QIAN
Chinese Journal of Behavioral Medicine and Brain Science 2010;19(5):401-404
Objective To observe the effect of buyanghuanwu decoction on expression of immunoreactive protein and mRNA of NMDA receptor 2B subunit in rats hippocampal with vascular dementia to investigate the mechanism of buyanghuanwu decoction. Methods One hundred and forty-four rats were randomly divided into 4 groups: sham-operated group, VD model group,nimodipine group and buyanghuanwu decoction treatment group. The rats models of vascular dementia were built up by four vessels occlusion method. VD rats were treated with in-tragastrical buyanghuanwu decoction suspension (50 pharmacognostic g·kg-1·d-1) and nimodipine suspension (20 mg·kg-1·d-1) for 30 days. The learning and memory abilities were evaluated by Morris water maze tests. The change of NR2B protein in hippocampal of each group of rats were measured with immunohistochemistry and Western blot techniques and the expression of NR2B mRNA in hippocampus were observed by real-time fluorescence quantitative PCR techniques. Results Water maze tests,compared with sham-operated group((24. 18 ± 7.90)s,(7.99 ±1.32)/min) ,the escape latency(51. 25 ±18.28)s to explore the extension and the average spatial probe number ((5. 26 ±0. 74)/min) reduced in VD model group (P < 0. 05). Buyanghuanwu decoction ((25.91 ±9.56)s,(7. 52 ± 1. 27)/min) had significantly improved the above-mentioned rat model of learning and memory performance (P<0.05). There was no significant difference among sham-operated group,nimodipine group and buyanghuanwu decoction treatment group (P>0. 05). Similarly,as compared rats with sham-operated group(0.71 ±0.13), (5887 ±501), the expression of NR2B protein (0. 33 ± 0. 06) and its mRNA(593 ±53) were apparently decreased in VD rats (P< 0.05). The expression of NR2B protein(0.66 ±0. 11) and its mRNA (5692 ±482) in neuron of hippocampus were increased by buyanghuanwu decoction compared with the model group (P < 0. 05), and no difference was discovered between sham operation group and nimodipine group (P > 0.05).Conclusions Buyanghuanwu decoction improves the learning and memory abilities in VD rats, the therapeutic mechanism was concerned with lessening the injury of neurons on CA1 field in hippocampus and promoted the expression of NR2B protein and its mRNA.