Inhibitory effect and mechanism of total flavonoids from Melicope pteleifolia on transplanted tumor of colorectal cancer in nude mice / 中国药房

OBJECTIVE To study the inhibitory effect and mechanism of total flavonoids from Melicope pteleifolia （TF-MPL） on transplanted tumor of colorectal cancer in nude mice. METHODS The transplanted tumor model of colorectal cancer was induced by injecting 0.2 mL colorectal cancer cell LoVo subcutaneously via the right armpit of nude mice. After successful modeling， nude mice were randomly divided into model group， 5-fluorouracil group （positive control， 10 mg/kg）， TF-MPL high- dose and low-dose groups （25， 12.5 mg/kg）； a normal group （normal saline containing 0.3% carboxymethyl cellulose sodium） without modeling was additionally set up， with 6 mice in each group. Each group was intraperitoneally injected with the corresponding drug solution/solvent for 21 consecutive days. The inhibitory rate of the transplanted tumor， liver and spleen index， and the levels of tumor necrosis factor-α （TNF-α） and interleukin-6 （IL-6） in serum were detected after the last medication； the morphological changes of tumor tissue were observed； immunohistochemical staining was used to detect protein expressions of Toll- like receptor 4 （TLR4） and nuclear factor-κB subunit p65 （NF-κB p65） in tumor tissue of nude mice. Western blot assay was used to detect protein expressions of TLR4， myeloid differentiation factor 88 （MyD88）， TNF receptor-associated factor 6 （TRAF6）， interleukin-1 receptor-associated kinase 1 （IRAK-1）， NF-κB p65 and caspase-3 in tumor tissue of nude mice. RESULTS Compared with the model group， TF-MPL high-dose group showed a significant decrease in tumor weight （inhibitory rate of 36.91%）， liver and spleen index， serum levels of TNF-α and IL-6 and protein expressions of TLR4， MyD88， TRAF6，IRAK-1 and NF- κB p65 （P＜0.05 or P＜0.01）； the expression of caspase-3 protein was increased significantly （P＜0.05）， and more tumor cell shrinkage and deformation， nuclear pyknosis and fragmentation were observed. CONCLUSIONS TF-MPL can significantly inhibit the growth of transplanted tumor of colorectal cancer in nude mice， the mechanism of which may be associated with reducing inflammatory response， inhibiting TLR4/MyD88/NF-κB signaling pathway， and promoting apoptosis in colorectal cancer cells.

The effect and mechanism of anwulignan on improving hepatic fibrosis in rats / 中国药房

OBJECTIVE To investigate the effect and mechanism of anwulignan on improving hepatic fibrosis in rats. METHODS Fifty SD rats were randomly divided into the normal group， model group， colchicine tablet group （0.1 mg/kg）， and anwulignan high-dose and low-dose groups （2.8 and 0.7 mg/kg）， with 10 rats in each group. Except for the normal group， all groups of rats were intraperitoneally injected with 50% CCl4 olive oil mixed solution to replicate the rat model of liver fibrosis. At the end of the modeling， rats in each group were given the corresponding drugs or distilled water intragastrically from the 9th week， once a day， for 4 weeks consecutively. During the experimental period， the general condition of the rats was observed； the liver index was calculated； the serum levels of alanine aminotransferase （ALT）， aspartate aminotransferase （AST）， superoxide dismutase （SOD） and malondialdehyde （MDA） were detected by colorimetric assay； the pathomorphology of the liver tissues and liver fibrosis were observed by HE staining and Masson staining； Western blot was used to detect the expression levels of phosphatidylinositol 3-kinase （PI3K）/protein kinase B （Akt） pathway and apoptosis-related proteins in liver tissues. RESULTS Compared with the normal group， the dietary amount of rats in the model group decreased， with sparse and disheveled fur， slow response， and a slower rate of weight growth or weight loss； the liver index was significantly increased （P＜0.01）； the serum levels of ALT， AST and MDA were significantly increased， and the SOD level was significantly decreased （P＜0.01）； HE and Masson staining showed that a large amount of fibrous proliferation was present in the liver tissues of the rats， and the collagen volume fraction was significantly increased （P＜0.01）； the protein expressions of PI3K， Akt， phosphorylated Akt and B-cell lymphoma （Bcl-2） were down-regulated significantly， while the protein expression of Bcl-2-associated X protein was increased significantly （P＜0.01）. Compared with the model group， the above indexes of the anwulignan high-dose and low-dose groups and the colchicine tablets group were all reversed significantly. CONCLUSIONS Anwulignan may reduce oxidative stress and inhibit hepatocyte apoptosis by activating the PI3K/Akt signaling pathway， and play the role of anti-hepatic fibrosis.

Mechanism of Improvement Effects of Fupi Rougan Granules on Hepatic Fibrosis Model Rats / 中国药房

OBJECTIVE：To study the mechanism of improvement effects of Fupi rougan granule （FRG）on hepatic fibrosis model rats. METHODS ：The rats were randomly divided into blank group ，model group ，Colchicine tablet group （chemical positive control ，0.2 mg/kg），Fuzheng huayu capsule group （TCM positive control ，0.415 g/kg），FRG low-dose ，medium-dose and high-dose groups （20，40，80 g/kg），with 10 rats in each group ，except for 11 rats in blank group and model group （one rat was used to judge whether the modeling was successful ）. Except for blank group ，other groups were given intraperitoneal injection of 50％ CCl4 olive oil solution and intragastric administration of 30％ ethanol to induce hepatic fibrosis model. After modeling ， administration groups were given relevant medicine intragastrically ；blank group and model group were given constant volume of normal saline intragastrically ，once a day ，for consecutive 4 weeks. After last administration ，morphology changes of liver tissue in rats were observed. The serum levels of HA ，LN，PCⅢ and Col Ⅳ in rats were detected ，and protein expression of Beclin- 1 and LC3-Ⅱin liver tissue were also determined. mRNA and protein expression of Akt ，AMPK，mTOR，p70S6K were detected in liver tissues of rats. RESULTS ：Compared with blank group ，the structure of hepatic lobules in the model group was disordered ，the proliferation of fibrous tissue was obvious ，and some pseudolobules were formed ；the serum levels of HA ，LN，PCⅢ and Col Ⅳ， the protein expression of Beclin- 1 and LC 3-Ⅱ in liver tissue as well as mRNA and protein expression of Akt ，AMPK，mTOR and p70S6K were increased significantly （P＜0.01）. Compared with model group ，the liver injury of rats in FRG groups was significantly relieved ，and the levels of the above indexes in serum and liver tissue （except for LN and PC Ⅲ in FRG low-dose group） were significantly reduced （P＜0.05 or P＜0.01）. CONCLUSIONS ：FRG can improve hepatic fibrosis in rats ，the mechanism of which may be associated with down-regulating the expression of autophagy associated protein and Akt/AMPK/mTOR/ p70S6K signaling pathway related protein.

Effect of plasma exchange in the treatment of severe hepatitis / 中国基层医药

Objective:To investigate the clinical effect of plasma exchange in the treatment of patients with severe hepatitis.Methods:From December 2011 to December 2018, 84 patients with severe hepatitis admitted to the Third People's Hospital of Linfen were selected, and they were divided into control group ( n=41) and observation group ( n=41)according to the random digital table method.The control group was treated with routine treatment, and the observation group was treated with plasma exchange at the same time.The therapeutic effect of the two groups was observed. Results:The total effective rate of the observation group was 73.17%(30/41), which was significantly higher than that of the control group[51.22%(21/41)] (χ 2=4.201, P<0.05). After treatment, the ALB, AST, ALT, TBIL levels in the control group were (36.74±4.25)g/L, (247.85±12.36)U/L, (214.57±10.14)U/L, (288.96±16.30)μmol/L, respectively, which in the observation group were (45.14±5.30)g/L, (162.65±8.30)U/L, (120.74±6.33)U/L, (241.74±15.02)μmol/L, respectively, the differences between the two groups were statistically significant( t=7.917, 36.642, 50.261, 13.641, all P<0.05). After treatment, the interferon gamma(IFN-γ), tumor necrosis factor alpha(TNF-α), interleukin 6(IL-6) levels in the control group were (318.96±92.15)ng/L, (334.74±102.58)ng/L, (65.89±6.33)ng/L, respectively, which in the observation group were (261.15±89.62)ng/L, (274.15±85.12)ng/L, (54.36±5.23)ng/L, respectively, the differences between the two groups were statistically significant( t=2.879, 2.910, 8.991, all P<0.05). There was no statistically significant difference in the incidence of adverse reactions between the two groups ( P>0.05). Conclusion:Plasma exchange in the treatment of severe hepatitis can improve the clinical therapeutic effect, improve its liver function, reduce the level of inflammatory cytokines, and has no adverse reactions.

Clinical multicenter study of carboprost methylate suppository for cervical ripening prior to diagnostic hysteroscopy / 中华妇产科杂志

Objective To evaluate the effectiveness of carboprost methylate suppository for cervical ripening before diagnostic hysteroscopy in premenopausal women. Methods From July 2014 to July 2015, 1614 women who were undergone diagnostic hysteroscopy in 12 hospitals were randomly assigned into study group(n=1209)and control group(n=405). The cases in study group were given 1 mg carboprost methylate suppository in vagina before hysteroscopy, the cases in control group were given 1 mg placebo. The extent of cervical ripening, the time of dilated cervix, pain scoring, incidence of drug side reactions after 24, 48, 72 hours, satisfaction degree of operators and patients, the time of hysteroscopy, incidence of complications between the two groups were observed and compared. Results (1) Mean cervical widths in the study and control groups were 6.11 ± 1.11 and 5.95 ± 1.11, and showed a significant difference(P=0.034);the percentage of women requiring cervical dilatation in study group was lower than the percentage in control group significantly [28.3％(342/1209)versus 34.6％(140/405), P=0.020].(2) The time of dilated cervix in study group was shorter than the time in control group significantly [(34 ± 25) versus(52 ± 49)s, P=0.028] for the patients whose mean cervical widths≤4.(3)There was no significant difference in pain scores between the two groups(P>0.05).(4)The incidence of side reactions 24, 48, 72 hours after operation were no significant difference between the two groups (P>0.05). (5) The satisfaction degree of operators and patients, the time of hysteroscopy, incidence of complications between the two groups were no singnifcant difference between the two groups (all P>0.05). Conclusion Application of carboprost methylate suppository by vagina before hysteroscopy is an effective and safe method of cervical ripening.

Effects of estrogen on the fibrosis process of intrauterine adhesions and the expression of forkhead box F2 / 解放军医学杂志

Objective To investigate the effect of estrogen on the fibrosis process of intrauterine adhesions and the expression of forkhead box F2 (FoxF2).Methods Primary human endometrial stromal cells (HESCs) were obtained by separation with 0.2％ collagenase Ⅰ digestion-mesh filtration-differential adherence,and identified by immunocytochemistry.HESCs affected with 10ng/ml transforming growth factor β1 (TGF-β1) for 48 hours.HESCs in model group were affected with 0,10-6,10-8,10-10 and 10-12mol/L estrogen,the expressions of smooth muscle actin alpha (α-SMA),Collagen I (COL Ⅰ) and FoxF2 were detected by quantitative PCR (qPCR) and Western blotting.Results HESCs with high purity and good activity were obtained by using 0.2％ collagenase Ⅰ digestion-mesh filtration-differential adherence separation method.Immunocytochemistry showed positive vimentin and negative cytokeratin 18 in HESCs.The results of qPCR and Western blotting showed that the mRNA and protein expression levels of α-SMA,COL Ⅰ and FoxF2 were higher in model group than in control group (P＜0.05),the model was built successfully.qPCR revealed that the mRNA expression levels ofα-SMA,COL Ⅰ and FoxF2 were significantly lower in 10-6,10-8 and 10-10mol/L estrogen groups than in model group (P＞0.05 in 10-10mol/L estrogen group,P＜0.05 in other groups),while in 10-12mol/L estradiol group,the expression levels of FoxF2 mRNA significantly decreased (P＜0.05),and of α-SMA and COL Ⅰ mRNA increased,but no significant difference were found (P＞0.05).Compared with the model group,the protein expression levels of α-SMA,COL Ⅰ and FoxF2 in 10-6,10-8 and 10 10mol/L estrogen groups decreased,but no significant difference was found (P＜0.05),while in 10-12mol/L estradiol group,the expression levels ofα-SMA protein increased (P＞0.05),and of COL Ⅰ and FoxF2 proteins decreased (P＜0.05).Conclusions The expression of FoxF2 in intrauterine adhesions is increased.Estrogen can reverse the fibrosis process of intrauterine adhesions in a certain range and inhibit the expression of FoxF2.

Predictive efficacy of different intrauterine adhesion (IUA) classification systems on the prognosis of patients with IUA / 解放军医学杂志

Objective To explore the effects of different intrauterine adhesion (IUA) classification systems on predicting the IUA prognosis.Methods One hundred cases were selected as the subjects in present study from those diagnosed with IUA and underwent surgery in Zhujiang Hospital of Southern Medical University from Jan.2010 to Jan.2017,and were followed up for two years.According to the actual situation,all patients were scored by March,AFS,ESGE and Chinese classification for comparing the effects of different IUA classification systems on predicting the pregnancy rate,live birth rate and effective rate within 2 years after surgery.Results ESGE classification had a good effect on predicting the postoperative live birth rate and effective rate,and a certain predictive effect on pregnant rate,with the area under curve (AUC) of 0.722,0.754 and 0.635,respectively.March classification had a certain effect on predicting the postoperative live birth rate and effective rate with AUC of 0.635,0.754,respectively,but had a poor effect on predicting pregnant rate.AFS classification and China classification had poor effect on predicting the IUA prognosis.Conclusion ESGE classification system is better than the other systems including March,AFS and Chinese classification,on predicting the IUA prognosis,but further verification in large sample size is still required.

Expression of Ki-67 and estrogen receptor in the uterus of mice with autoimmune premature ovarian failure induced by peptide zona pellucida 3 / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the histomorphology and the expressions of the proliferation marker Ki-67 and estrogen receptor in the uterus of mice with autoimmune premature ovarian failure (POF) induced by zona pellucida 3 peptide (pZP3).</p><p><b>METHODS</b>Autoimmune POP models were established in 20 female BALB/c mice (7-8 weeks old) by immunization with pZP3 and another 20 mice served as the control group. The POP models were verified by vaginal cytology, serum sex hormones, ovary histomorphology and ZP3 antibody immunohistochemistry. The histomorphology and expressions of Ki-67, estrogen receptor α and estrogen receptor β in the uterus of the mice were detected.</p><p><b>RESULTS</b>Autoimmune POP models were established successfully in 80% of the mice at 8 weeks after the immunization. Compared with those in the control group, the mice in the model group showed a smaller volume of the uterus, thinner endometrium and a reduced number of glands. The luminal epithelial cells, glandular epithelial cells and stromal cells in the uterus of the model mice all presented with a lower expression of Ki-67 than those in the control group, and Ki-67 translocation from the nuclei to the cytoplasm was found in the model group. The luminal epithelial cells, glandular epithelial cells and stromal cells showed positive ERα immunoreactivity in the model group but not in the control group. No obvious ERβ expression was found in the uterus in either of the groups.</p><p><b>CONCLUSION</b>pZP3 can induce autoimmune POP, cause suppressed proliferation of the endometrial epithelial cells and stromal cells, and reduce the cellular expression of ERα in the uterus of mice.</p>

Effect of TGF-β1on the expression of MMP-9 in mouse endometrial stromal cells / 重庆医学

Objective To investigate the effect of transforming growth factor-β1(TGF-β1) on the expression of matrix metal-loproteinase-9(MMP-9) in cultured mouse endometrial stromal cells .Methods After separation and purification ,the mouse endom-etrial stromal cells were cultured with different concentrations of TGF-β1 ,and the final concentrations were 0 .1 ,2 .5 ,5 .0 ,10 .0 , 20 .0 ng/mL ,respectively .ELISA method was used to detect the content of MMP-9 in the the culture medium after 48 h culture . Results Compared with the control group ,the expression of MM P-9 in mouse endometrial stromal cells in the experimental groups increased with the TGF-β1 concentration elevation ,and the difference was statistically significant (P < 0 .01) .Conclusion In the mouse endometrial stromal cell system ,TGF-β1 may be involved in the regulation of extracellular matrix metabolism through regu-lating the expression of MMP-9 .

Bone marrow mesenchymal stem cell transplantation for treatment of cis-platinum-induced ovarian damage / 中国组织工程研究

BACKGROUND:Chemotherapy drugs can damage the ovarian function in women of childbearing age, and even lead to premature ovarian failure. Therefore, to improve and restore the ovarian function in patients has become an important issue. OBJECTIVE:To explore the therapeutic effect and feasibility of bone marrow mesenchymal stem cel therapy against chemotherapy-induced ovarian damage. METHODS:Rat models of chemotherapy-induced premature ovarian failure were established, and injected with PKH26-labeled bone marrow mesenchymal stem cels. At 15, 30, 45, 60 days after cel transplantation, five rats were selected respectively to detect folicle-stimulating hormone and estradiol levels, and then, the rats were kiled to take the right ovary for pathological examination. The number of ovarian folicles was detected under light microscope. At 30 days after cel transplantation, another two rats were selected to mate with male rats to observe the difference in the reproductive activity. RESULTS AND CONCLUSION:Four of 22 rats (18%) gradualy recovered their estrous cycle after cel transplantation, with the decreased folicle-stimulating hormone level and increased estradiol level. Moreover, the number of folicles was reduced. Al of these indicated that the ability to have children in rats was not damaged.These experimental findings suggest that bone marrow mesenchymal stem cels can partialy improve the ovarian function of rats under chemotherapy.

Correlation between interleukin-21 and formation of intrauterine adhesions in rats / 医学研究生学报

Objective Interleukin-21(IL-21) is likely to contribute to the development of liver fibrosis, but up to now, no study has been reported on the relationship between IL-21 and intrauterine adhesions ( IUA) .This study aimed to establish a rat model of IUA induced by mechanical injury and lipopolysaccharide (LPS) infection (dual injury), determine the expression level of serum IL-21, and confirm the association of serum IL-21 with the formation of IUA. Methods Forty healthy female SD rats were randomly divided into four groups of equal number,control, mechanical injury, LPSinfection, and dual injury.At 7 days after IUA modeling,uterine tissue-swere collected from all the animals for observation of the endometrial glands,detection of the degree of IUA by Masson staining, measure-ment of the serum IL-21 level by radioimmunoassay, and analysis of the correlation between the number of endometrial glands and the de-gree of fibrosis. Results The number of endometrial glandswas significantly smallerin the dual injury group (3.59±1.20) than in the mechanical injury (11.66±2.34) and LPSinfection group(11.59±1.47)(P<0.05), while the proportion of fibrosis area wassignificantly higher in the former group(0.65±0.03) than in the lattertwo(0.30±0.07 and 0.32±0.08)(P<0.05).The level of serum IL-21 was signifi-cantly increased in the dual injury group ([286.21±27.80]pg/mL) as compared with those in the control ( [ 118.65 ±22.55 ] pg/mL ) , mechanical injury([176.20±19.05]pg/mL), and LPS infection group ([187.98±16.51]pg/mL) (P<0.05), with a positive correlation be-tween the IL-21 level and theproportion of fibrosis area ( r=0.271, P<0.05) . Conclusion A rat model ofIUAwas successfully established by mechanical injury and lipopolysaccharide (LPS) infection.The evi-dent increase of serum IL-21 in the IUA model was positively correlated with the percentage of fibrosis area, suggesting that IL-21 may be involved indirectly in the formation of IUA.

Lentiviral vector-mediated short hairpin RNA targeting survivin inhibits abdominal growth of human endometrium xenograft in nude mice / 南方医科大学学报

<p><b>OBJEVTIVE</b>To investigate the inhibitory effect of lentiviral vector-mediated short hairpin RNA targeting survivin (LV-survivin shRNA) on the growth of human endometrium xenograft in the abdominal cavity of nude mice.</p><p><b>METHODS</b>The endometrium xenografts from 8 women with endometriosis were injected into the peritoneal cavities of 45 nude mice. The mice were then randomly assigned to receive intraperitoneal injection of LV-survivin shRNA, pGCL-NC-GFP (negative control) or PBS (blank control). Two weeks later, the number and morphometry of endometriotic lesions were quantified and the expression of survivin protein were detected by immunohistochemistry.</p><p><b>RESULTS</b>The formation of endometriotic lesions was significantly suppressed in mice receiving LV-survivin shRNA injection as compared with those in the two control groups (P/0.001). The mice in LV-survivin-shRNA group showed significantly down-regulated expression levels of survivin protein compared with those in the negative and blank control groups, presenting also necrosis in the endometriosis-like lesions in microscopic observation.</p><p><b>CONCLUSION</b>Lentiviral vector-mediated shRNA can effectively inhibit the expression of survivin in human endometrium xengrafts and suppress the formation and growth of endometriotic lesions in the abdominal cavities of nude mice.</p>

Effective administration model for gestational diabetes mellitus in the community / 实用医学杂志

Objective To investigate the effective administration model for gestational diabetes mellitus (GDM) in the community. Methods In a prospective study, the 75 g oral glucose tolerance test (OGTT) was performed in 4 713 resident pregnant women over 20 years old who received antenatal care in a general hospital or a special hospital from Sep. 2011 to Aug. 2012. Five hundred and thirty-three pregnant women were diagnosed as GDM, 198 patients who labored in a general hospital were enrolled in thegroup A , and the rest who labored in a special hospital were enrolled in the group B. 198 cases with non-GDM were enrolled in the group C. Results The incidence of GDM during this study period was 11.3%. The maternal age , gestationl weeks and OGTT results of patients in the three groups were significantly different (P 0.05). However, significant differences were found in the incidence of postpartum hemorrhage and cesarean section. The incidence of postpartum hemorrhage increased significantly in the patients of group B (χ2= 7.156, v = 2, P = 0.028). The incidence of cesarean section increased significantly in the patients of group A (χ2= 63.592, v = 2, P = 0.000). Conclusion Establishing an effective administration model for gestational diabetes mellitus in the community could control the incidence of GDM associated complications.

Apoptosis of human primary ovarian granulose cells infected with lentivirus carrying bcl-2 gene / 中国组织工程研究

BACKGROUND: Lentivirus can infect divided and undivided cells. It remains uncertain whether the lentivirus can successful y infect primary ovarian granulosa cells. OBJECTIVE: To investigate infecting ratio and cel apoptosis of lentivirus carrying bcl-2 gene in primary human ovarian granulose cells cultured in vitro. METHODS: The lentiviral vector carrying bcl-2 gene was constructed using molecular biology, and packaged into lentivirus with high titer. The resulting recombinant lentivirus carrying bcl-2 genes were then used to infect primary human ovarian granulosa cells in vitro at different multiplicity of infection, 10, 50, 100, 200, and 400. Infection efficiency and cel proliferation were observed at 24, 48, 72, and 96 hours fol owing infection. Cel apoptosis was detected by flow cytometry, and bcl-2 gene transcription was assessed using reverse transcription PCR. RESULTS AND CONCLUSION: Primary human ovarian granulosa cells adhered at 24 hours, and exhibited polygon- or fusiform-shape and colony-like growth. When multiplicity of infection was 100, cel appearance and growth remained unchanged, and infection efficiency was high, which reached the peak up to 72 hours. Moreover, the positive rate was up to 60% in granulosa cells. Lentivirus carrying bcl-2 gene could increase expression of Bcl-2 protein and inhibit apoptosis of primary ovarian granulosa cells.

Effect of cervical conization through hysteroscopy for cervical intraepithelial neoplasia III / 南方医科大学学报

<p><b>OBJECTIVE</b>To evaluate the therapeutic effect of cervical conization through hysteroscopy in the treatment of cervical intraepithelial neoplasia (CIN) III.</p><p><b>METHODS</b>Seventy-four patients with CIN III underwent cervical conization through hysteroscopy (TCRC group), and 65 received cold knife conization (CKC group). The operating time, volume of blood loss, concordance rate with pathology, recurrence rate, rate of cervix adhesion and pregnancy rate were compared between the two groups.</p><p><b>RESULTS</b>The operating time, mean blood loss, cure rate, and recurrence rate were 15.1∓3.2 min, 12.5∓1.8 ml, 94.6%, and 5.4% in TCRC group, respectively, as compared with those of 25.8∓3.8 min, 21.6∓2.4 ml, 81.5%, and 18.5% in CKC group, all showing significant differences between the two groups (P<0.05).</p><p><b>CONCLUSION</b>Compared with CKC, TCRC has such advantages as less blood loss, shorter operating time, more accurate lesion localization, fewer complications, higher cure rate, and lower recurrence rate without significant adverse effect on pregnancy.</p>

Protective effect of lentivirus-mediated Bcl-2 gene transfection against phosphoramide mustard-induced apoptosis of human ovarian granulosa cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effect of lentivirus-mediated Bcl-2 gene transfection in protecting human primary ovarian granulosa cells against phosphoramide mustard (PM)-induced apoptosis.</p><p><b>METHODS</b>Granulosa cells were isolated from the follicle fluid of women undergoing in vitro fertilization and embryo transfer. The lentiviral vectors carrying Bcl-2 gene (pGC-FU-Bcl-2) and enhanced green fluorescence protein (pGC-FU-EGFP) were constructed and packaged into high-titer lentiviruses. The resulting recombinant lentivirus carrying Bcl-2 and EGFP genes or the empty vector were used to infect the primary human ovarian granulosa cells, followed by addition of PM in the cell culture, with untreated granulosa cells as the control. The cell apoptosis was detected by Annexin V and Hochst 33258 staining, and the expression of Bcl-2 protein was assessed using Western blotting.</p><p><b>RESULTS</b>The control granulosa cells showed an apoptotic rate of (1.93±0.28)%. The cells infected with pGC-FU-Bcl-2 prior to PM exposure had a apoptotic rate of (6.99±10.55)%, significantly higher than that of the control cells, but significantly lower than that of the cells with PM exposure only and those infected with the empty vector before PM exposure (P<0.05). The expression of Bcl-2 was the highest in the cells infected with pGC-FU-Bcl-2 prior to PM exposure (P<0.05).</p><p><b>CONCLUSION</b>Lentivirus-mediated Bcl-2 gene transfection can protect human ovarian granulosa cells against PM-induced apoptosis by upregulating Bcl-2 protein expression.</p>

Effect of lentiviral vector-mediated short hairpin RNA targeting survivin against endometriosis-like lesions in chick embryo chorioallantoic membrane / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the inhibitory effect of lentiviral vector-mediated short hairpin RNA targeting survivin (LV-survivin shRNA) on angiogenesis and growth of endometriosis-like lesions in chick en embryo chorioallantoic membrane.</p><p><b>METHODS</b>Eutopic endometrium from women with endometriosis was transplanted onto the non-vascular region of (CAM), where LV-survivin shRNA was delivered subsequently. The angiogenesis and the growth of endometriosis-like lesions in the CAM model were evaluated.</p><p><b>RESULTS</b>The angiogenesis and formation of endometriosis-like lesions were significantly suppressed in the CAM model by treatment with LV-survivin shRNA in comparison with those in the untreated CAM models and models treated with empty LV or DMEM (P<0.001). LV-survivin shRNA also caused a significantly higher cell apoptotic rate in the endometriosis-like lesions than the other treatments (P<0.001) and induced necrosis in the lesions.</p><p><b>CONCLUSION</b>LV-survivin shRNA can effectively inhibit angiogenesis induced by the eutopic endometrium and markedly suppress the formation of endometriosis-like lesions in the CAM model.</p>

Application of micro-computed tomography-based 3D visualization in diagnosis and treatment of cervical cancer / 南方医科大学学报

<p><b>OBJECTIVE</b>To define the optimal development time of micro-computed tomography (micro-CT) venography in cervical cancer patients and establish 3D CT-based digital pelvic model of the patients.</p><p><b>METHODS</b>Thirty patients with cervical cancer stratified by FIGO surgical staging underwent micro-CT scanning of the arterial phase and the venous phase with a delay time of 70, 90 and 120 s. The images were interpreted independently by two experienced radiologists to define the optimal development time in venous phase and establish the imaging diagnosis. Based on the pelvic CT scan data, we segmented the images using the abdominal medical image-3D visualization system followed by 3D image reconstruction to establish the 3D digital pelvic model using FreeForm Modeling System to modify the reconstructed images.</p><p><b>RESULTS</b>The optimal images were obtained by scanning with a 90-sec delay time. Micro-CT was not sensitive to IB1 phase or earlier phases (1/5), but efficient in advanced stages (≥IB2 phase). In our cases, 25 were diagnosed by micro-CT with a diagnostic accuracy of 64%. Based on these CT data, the pelvic 3D model covering the main organs, vessels, cervical neoplasm, and supplying vessels of the tumor in the pelvic cavity were successfully reconstructed to allow spatial observations of the uterus and the neighboring organs.</p><p><b>CONCLUSION</b>The 3D digital pelvic model reconstructed provides a means for staging cervical cancer and facilitates further surgical simulation studies.</p>

Stereological Study on Syncytial Cell of Human Placenta in Early-onset Severe Preeclampsia / 实用妇产科杂志

Objective:The characteristics and changes of the three-dimensional micro structure of human placenta in Early-onset severe preeclampsia (EOSP) were quantified, and the relationship of morphologic features and the poor fetal prognosis was explored. Methods:10 patients were recruited in EOSP group, 10 normal blood pressure premature pregnancy were as control group. Ultrathin sections were observed by elec-tron microscope. The Vv,Sv,Nv of plancenta were measured by stereology method. These parameters were compared between the two groups. Results:①In EOSP group, the miorovillus at the bottom of syncytial was sparse and swollen. The mitochondria and endoplasmic reticulum were swollen and dilated. The deposition of lipid granules were increased.②The Vv, Sv of mitochondrial and endoplasmic reticulum in EOSP group were significant augmented than control groups( P<0.01 ). There was no significant difference of Nv in mitochon-drion and endoplasmic reticulum between two groups ( P >0.05). ③The Vv,Sv, Nv of microvilli in EOSP group were smaller than those in control group ( P < 0.01 ). Conclusions. In EOSP, the microstructure of placenta is impaired, thus the synthesis and transportation function of placenta were affected, so fetus could be injured subsequently.

Cloning and sequencing of rat bcl-2 gene riched guanine and cytosine / 中国组织工程研究

BACKGROUND: Excessive apoptosis of ovary granulosa cell is a dominant cause of premature ovarian failure and bcl-2 gene is able to inhibit cell apoptosis. But studies demonstrate that,the guanine and cytosine (GC) content reaches 60% in the rat bcl-2 gene sequence. This gene cannot be amplified using routine polymerase chain reaction method. OBJECTIVE: To clone and identify the bcl-2 gene riched GC. DESIGN,TIME AND SETTING: Open experiment was finished in the Laboratory of Medicine and Molecular Biology,Life Science School of Sun Yat-sen University from May to December in 2007. MATERIALS: Wistar rats were purchased from Experimental Animal Center of Sun Yat-sen University. Ecoli DH5?was preserved by Laboratory of Medicine and Molecular Biology,Life Science School of Sun Yat-sen University; pMD18-T vector was purchased from Takara Biotechnology (Dalian) Co.,Ltd. METHODS: The bcl-2-cDNA,in which GC accounted for 60.6%,was obtained by modified reverse transcription-polymerase chain reaction from kidney tissue of Wistar rats,and was cloned into vector-pMD18-T. Characterizations and sequencing of the pMD18-T-bcl-2 were carried out by polymerase chain reaction screening of individual bacterial colonies. MAIN OUTCOME MEASURES: Cloning and purification of bcl-2 gene cDNA; results after connecting bcl-2 gene cDNA to pMD18-T vector and transducting Ecoli DH5?,identification of positive clone and results of sequencing. RESULTS: The bcl-2 gene was identified by the clone and DNA sequencing. DNA sequence analysis was consistent with Genebank sequence,with a 99% homology. CONCLUSION: The gene riched GC is difficult to be amplified,bcl-2-cDNA can be cloned and constructed into cloning vector pMD18-T successfully by the efficient technique for other genes riched GC.