OBJECTIVE:To study the effects of Shenfu qiangxin(SFQX)pills on the expression of autophagy-associated pro-tein LC3b and pro apoptotic gene Bax in myocardial cells of rats with cardiorenal syndrome (CRS). METHODS:Rats were ran-domly divided into sham operation group(water),model group(water),positive control group(Captopril tablets 2.3 mg/kg)and SFQX pills high-dose,medium-dose and low-dose groups [13.2,6.6,3.3 g(crude drug)/kg],with 10 rats in each group. CRS mod-el was induced in those groups by abdominal-aortae-constriction+acute renal ischemia reperfusion injury except for sham operation group;and they were given relevant medicine intragastrically 8 week after operation,once a day,for consecutive 4 weeks. Plasma contents of Cr and ALD,the protein expression of LC3b and Bax in myocardial tissue of rats were detected 24 h after last medica-tion;ventricular index was calculated,and morphological change of myocardial tissue was observed. RESULTS:Compared with sham operation group,the plasma contents of Cr and ALD,ventricular index and the protein expression of LC3b in myocardial tis-sue increased significantly in model group (P<0.05 or P<0.01);and myocardial cell suffered from endochylema red deletion, myocardial cross striation disorder,intercellular space fibrosis aggravation and so on. Compared with model group,the plasma con-tents of Cr and ALD(except for positive control group)and the protein expression of LC3b and Bax in myocardial tissue decreased significantly in positive control group and SFQX pills high-dose group(P<0.05 or P<0.01);myocardial pathological change was improved;the ventricular index decreased significantly in SFQX pills low-dose and medium-dose groups (P<0.05). CONCLU-SIONS:SFQX pills can decrease the plasma contents of Cr and ALD,inhibit myocardial cell autophagy and apoptosis in CRS rats.

The culture conditions of Saccharomyces cerevisiae sp.strain by 1.1 b were optimized for the production of D-(-)-mandelate dehydrogenase which is useful for the asymmetric bioreduction of benzoylformate to form D-(-)-mandelate.The optimum medium(per liter)consistes of 60 g peptone,30 g maltose, 0.5 g MgSO_4,0.01 g ZnSO_4,1.0 g KCl.After optimization of the culture medium,the enzyme production in shake flasks is enhanced from 2.56 to 20.21 U/L.The optimum fermentation conditions were determined as follows:medium volume 100 mL(i.e.,40%for a 250-mL shake flask),pH 6.5,inoculum size 10%,temperature 30℃,and cultivation time 25 h.

OBJECTIVETo study the inhibitory effect and mechanism of Ganoderma lipsiense extract (GLE) on the growth of triple-negative breast cancer (TNBC) cell line MDA-MB-231-HM in a mouse model.

METHODSThe mouse model of TNBC was established by subcutaneous injection of 1.5 x 10(6) of MDA-MB-231-HM cells into BALB/c-nu mouse. Twenty successfully modeled mice were divided into the GLE group and the negative control group according to random digit table, 10 in each group. GLE (0.2 mL 100 mg/mL) was peritoneally injected to mice in the GLE group, while equal dose of normal saline was peritoneally injected to mice in the negative control group. The medication was administered once per 3 days and discontinued after 45 days. The CD34 expression was detected using immunohistochemical assay for counting microvessels. Meanwhile, expressions of thrombospondin 1 (TSP-1) and cyclin D1 were detected using immunohistochemical assay.

RESULTSThe average weight was obviously lower in the GLE group than in the negative control group [(0.33 ± 0.16) g vs (0.68 ± 0.37)g, P < 0.05]. The tumor inhibition rate was 51.4% in the GLE group. The volume of transplanted tumor was obviously lesser in the GLE group than in the negative control group (P < 0.05). Results of immunohistochemical staining showed, the microvessel density (MVD) under every field was (20.7 ± 2.1), TSP-1 positive cell count was (66.2 ± 9.2), cyclin D1 positive cell count was (33.8 ± 16.4) in the GLE group, and they were 34.0 ± 2.0, 24.0 ± 6.6, and 168.2 ± 32.6, respectively in the negative control group. There was statistical difference in all indices between the two groups (P < 0.05).

CONCLUSIONGLE could inhibit malignant proliferation of tumor cells by suppressing angiogenesis of blood vessels in tumor tissues and regulating cell cycles, thereby inhibiting TNBC.

Animals ; Biological Products ; pharmacology ; Cell Line, Tumor ; Cyclin D1 ; metabolism ; Disease Models, Animal ; Ganoderma ; chemistry ; Humans ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Microvessels ; Neoplasm Transplantation ; Neovascularization, Pathologic ; prevention & control ; Random Allocation ; Thrombospondin 1 ; metabolism ; Triple Negative Breast Neoplasms ; drug therapy

Objective: To establish the cardio-renal syndrome (CRS) model by coarctation of abdominal aorta (CAA) with renal ischemia reperfusion injury (RIRI), and to observe the mRNA expression of pro-renin receptor [(P)RR] in experimental rats. Methods: A total of 42 Wistar rats were randomly divided into 4 groups: Sham group, CAA group, RIRI group and CAA+RIRI group.n=10 in each group, 2 rats died during the modeling and all animals were treated for 16 weeks. Blood levels of BNP, creatinine (Cr), urea nitrogen (BUN), the activity of rennin, the contents of angiotensin-I (AT-I), AT-II and aldosterone were examined by laboratory test. The diastolic end inter-ventricular septum thickness (DEIVST), DELVPT, LVEF, ventricular weight index (VWI) and cardiac weight index were detected by small animal echocardiography. The histological changes of myocardium and kidney tissue were measured by HE staining, and the mRNA expressions of pro-renin receptor in myocardium and kidney tissues were measured by RT-PCR. Results: Compared with Sham group, blood levels of BNP were increased in the other 3 groups,P<0.05; compared with CAA group, CAA+RIRI group had increased levels of Cr and BUN,P<0.01; compared with Sham group and RIRI group, CAA+RIRI group showed increased blood level of aldosterone,P<0.05. Compared with CAA group, CAA+RIRI group presented increased rennin activity,P<0.05. Blood levels of AT-I and AT-II were not signiifcantly increased among 3 operation groups,P>0.05. Compared with CAA group, CAA+RIRI group had more obvious changes of DEIVST and LVEF,P<0.01. Compared with RIRI group, CAA+RIRI group had more obvious ventricular hypertrophy, higher VWI and cardiac weight index, allP<0.05. HE staining presented that CAA+RIRI group had broadening of myocardial cell bundle space, decreased left renal index, severe tubular atrophy and partial glomerular atrophy. RT-PCR demonstrated that compared with Sham group, the mRNA expressions of pro-renin receptor in myocardium and kidney tissues were decreased in the other 3 groups. Conclusion: Combined CAA+RIRI method may damage the cardial and renal tissues at the same time which was more severe than either CAA or RIRI. While CAA+RIRI model has better controllability and higher consistency that provides a methodological reference for pro-renin receptor in treating CRS in experimental rat’s model.

Objective To determine the changes of interleukin-12(IL-12) and IL-12 mRNA in gastric mucosa of children with helicobacer pylori (Hp) infection,and to study the effects of Hp infection on the expression levels of IL-12 and IL-12 mRNA,and to evaluate its possible roles in the pathogenesis of gastric mucosal inflammation in Hp related gastroduodenal diseases.Methods Biopsy specimens were taken from the antral mucosa on endoscopy in patients with or without Hp infection, which were diagnosed by urease test and Giemsa staining. The expression levels of IL-12 and IL-12 mRNA in gastric mucosa were determined by ELISA and RT-PCR.Results Inflammation of gastric antral mucosa was more severe in Hp-positive mucosa .The expression levels of IL-12 and IL-12 mRNA in Hp-positive mucosa were (66.42?15.15) ng/g and (59.21?15.03)%,which were more than those in (Hp-negative )(22.22?8.79) ng/g and (17.94?7.39)%(P

Objective Establish rabbit VX2 soft tissue tumor model,and treat it with percutaneous ethanol injection(PEI)under the guidance of magnetic resonance imaging.Make ready for the therapeutic evaluation with functional magnetic resonance imaging. Methods Fifteen healthy New Zealand white rabbits were included in this study.0.2 mL tumor tissue suspensions were injected into the rabbits’posterior limb.14 days later,all rabbits were underwent conventional MRI examination.PET were performed to all the tumors under the guidance of MRI in the next day of the examination.T2 WI was used as guidance and monitoring means.MR com-patible puncture needle with lateral hole was stabed into the lesion center,and inj ected anhydrous ethanol according to the volume of tumors’diameter (1 mL/cm )slowly.the tumors signal characteristics,morphological feature and pathological feature were ob-served pre and post-operation.Results All of the 1 5 rabbits were established soft tissue tumor model successfully;the success rate is 100%.The tumors were oval or round,3-4 cm in diam.MRI scanning showed low signal on T1 WI and high signal on T2 WI be-fore PEI.PEI was performed to all the tumors under the guidance of MR successfully with 3.5 mL ethanol injected into the tumors in average.T2 WI could monitor the ethanol in dispersion and distribution within the tumors clearly.Histologically,tumors were composed of large,uniform,oval/round cells arranged in solid nests which was intensive in the periphery of tumors.Necrosis tissue was apparent in the center of the tumors.10 days after operation,most tissue in the periphery of tumors was coagulative necrosis , only a few tumor cells left.Ranges of necrosis in the tumors center were obviously increased compared with pre-operation.Conclusion Rabbit VX2 tumor of soft tissue model is suitable for the therapeutic evaluation of tumor .It is an animal model which has the characteristic of simple to operate and high rate of suc-cessful.MR T2 WI can monitor the ethanol in dispersion and distribution within the tumors clearly.It is a good guidance and monitoring imaging method of tumor ablation.

OBJECTIVETo investigate the correlation of bcl-2 and Bax protein with nuclear matrix in glioblastoma cell line U87 as well as the effect of EGFR-cDNA transfection on the expression of bcl-2 and Bax in U87 cells.

METHODSThe correlation of bcl-2 and Bax protein with nuclear matrix in glioblastoma cell line U87 was studied by confocal microscopy and Western blot. The expression of bcl-2 and Bax in EGFR-cDNA transfected and untransfected glioblastoma cell lines was studied by Western blot.

RESULTSConfocal microscopic images showed that bcl-2 protein was localized in the periphery of the nuclear matrix and Bax in the nuclear matrix. A 26 kDa bcl-2 band and a specific band of Bax at about 66 000 were detected in nuclear matrix proteins by western blot. The expression of bcl-2 was lower but that of Bax was higher in EGFR-cDNA transfected cells than the control.

CONCLUSIONBcl-2 and Bax, being nuclear matrix associated proteins, are probably involved in the EGFR-cDNA induced malignant conversion of glioblastoma cells by introducing EGFR cDNA into the tumor cells.

Apoptosis ; Cell Line, Tumor ; Glioblastoma ; pathology ; Humans ; Nuclear Matrix ; chemistry ; Proto-Oncogene Proteins c-bcl-2 ; analysis ; physiology ; Receptor, Epidermal Growth Factor ; genetics ; physiology ; Transfection ; bcl-2-Associated X Protein ; analysis ; physiology

OBJECTIVETo study the effect of dihydroartemisinin (DHA) combined irradiation on the apoptosis of human lung cancer GLC-82 cells and to study its mechanism.

METHODSThe growth inhibition rate of GLC-82 cells acted by different concentrations DHA was detected using MTT assay at 24, 48, and 72 h, respectively. Clone forming test was used. With multi-target single-hit model, the radiosensitization effect was assessed by calculating sensitizing enhancement ratio (SER).The effect of DHA combined irradiation on the apoptosis of GLC-82 cell cycle distribution and apoptosis were measured by flow cytometry. The protein expression of p53, p21, Bcl-2, and Bax were detected by Western blot.

RESULTSDifferent concentrations DHA (4, 8, 16, 32, 64, and 128 μg/mL) had cytotoxicity on GLC-82 cells. The IC50 for 24, 48, and 72 h was 38.25,20.58, and 10.36 μg/mL, respectively, in obvious dose- and time-dependent manner. The growth inhibition rate was more significantly increased than that of the blank control group (P < 0.01, P<0.05). DHA had sensitization enhancement effect on GLC-82 cells, with SER of 1.4. DHA combined irradiation could obviously change the structure of GLC-82 cells cell cycle and induce apoptosis (with the apoptosis rate of 21.5%), which was significantly different from that of the blank control group (P < 0.05). Western blot showed the expression of p53 and p21 protein could be increased by DHA combined irradiation, and the expression of Bcl-2 protein down-regulated (P <0.01, P <0. 05).

CONCLUSIONSDHA had stronger cytotoxicity and radiosensitization on GLC-82 cells. Its mechanisms might lie in making the arrest of GLC-82 cells' growth at G0/G1 phase, decreasing the ratio of cells at S phase, restoring the function of p53, decreasing the expression of Bcl-2 protein, and inducing apoptosis in GLC-82 cells.

Apoptosis ; drug effects ; Artemisinins ; pharmacology ; Cell Cycle ; drug effects ; Cell Proliferation ; drug effects ; Down-Regulation ; drug effects ; Flow Cytometry ; Humans ; Lung Neoplasms ; metabolism ; Neoplasm Proteins ; metabolism ; Radiation-Sensitizing Agents ; pharmacology ; Tumor Cells, Cultured ; bcl-2-Associated X Protein ; metabolism

OBJECTIVETo explore the significance and the role of the p53 gene mutation in the exon 4 to 8 in keloid fibroblasts.

METHODSTissue samples from twelve patients with keloid and twelve hyperplastic scar respectively were harvested for in vitro culture of fibroblasts, and normal skin samples from the same patients were employed as the control. Polymerase chain reaction-based single-strained conformational polymorphism (PCR-SSCP) and DNA sequencing were employed to detect p53 gene mutations of the fibroblasts.

RESULTSThe points and frameshift mutations in the exon 4, 5, 6, 7 of p53 gene were identified in 9 of the 12 keloid tissue samples. No p53 gene mutation was detected in all hyperplastic scar and normal skin samples.

CONCLUSIONp53 gene mutation might play an important role in the formation and development of keloids.

Female ; Fibroblasts ; metabolism ; Genes, p53 ; Humans ; Keloid ; genetics ; Male ; Mutation

Carcinoma, Adenosquamous ; complications ; diagnosis ; surgery ; Cystectomy ; Humans ; Male ; Middle Aged ; Prostatectomy ; Prostatic Neoplasms ; complications ; diagnosis ; surgery ; Urinary Bladder Neoplasms ; diagnosis ; secondary ; surgery