Animals ; Arteries ; drug effects ; physiology ; Blood Pressure ; drug effects ; Epimedium ; Flavonoids ; pharmacology ; Rats ; Rats, Inbred SHR ; Rats, Wistar

This study examined the role of PI3K/Akt pathway in radiosensitization of DNA damage of cervical carcinoma cells. The 50% inhibition concentration (IC(50)) of cisplatin and docetaxel in HeLa cells was detected by Mono-nuclear cell direct cytotoxicity assay (MTT) in vitro. HeLa cells were treated by cisplatin/docetaxel of 10 percent of IC(20) alone or combined with LY294002 for 24 h, and then radiated by different doses of X-ray. The cell survival ratio was obtained by means of clone formation. One-hit multi-target model was fitted to the cell survival curve to calculate dose quasithreshold (Dq), mean lethal dose (D(0)), 2Gy survival fraction (SF(2)) and sensitization enhancement ratio (SER). The pAkt and total Akt expression was detected by Western blotting and DNA damage by neutro-comet electrophoresis. The HeLa cells were randomly divided into 7 groups in terms of different treatments: Control; radiation treatment (RT) group; LY294002+RT group; cisplatin+RT group; docetaxel+RT group; LY294002+cisplatin+RT group; LY294002+docetaxel+RT group. The apoptosis ratio of each group was measured by flow cytometry. The results showed that docetaxel and cisplatin significantly enhanced the phosphorylation of Akt in radiation-treated HeLa cells. The Dq, D(0) and SF2 in LY294002-contained groups were lower than those in docetaxel or cisplatin+RT group. The SER in the LY294002+docetaxel+RT group was 1.35 times that of the docetaxel+RT group, and that in the LY294002+cisplatin+RT group 1.26 times that of the cisplatin+RT group. The Comet electrophoresis showed that tail distance in the LY294002+cisplatin+RT group or LY294002+docetaxel+ RT group was longer than in the cisplatin+RT group or docetaxel+RT group. The apoptosis ratio in the LY294002+cisplatin+RT group or LY294002+docetaxel +RT group was higher than in the cisplatin+RT group or docetaxel+RT group. It was concluded that inhibiting PI3K/Akt pathway can increase the effect of docetaxel and cisplatin on the radiosensitivity of HeLa cells and DNA damage resulted from radiation.

The aim of this research is to refine the protocol of purification of SA and identify the character of SA. By utilizing the cold-denaturing method, most of other kinds of protein were screened out and SA was purified from the fermentation broth of L-183 by using the refined affinity chromatography method. The rate of recollection was checked to be 75%～85%. By identification, it is indicated that the molecular weight of self-made SA was 74.5kD, the biotin-combining number 3.2, the activity 11.2u/mg, the pI around 7.4. So, the essential characters of SA are same as described by documents.

OBJECTIVETo observe the effect of Bushen Tiaochong Recipe (BTR) on proliferation and secretion of ovarian granulocyte (OGC) cultured with excessive androgen by serum pharmacologic technique.

METHODSFollicular stimulating hormone (FSH) contained and BTR contained (high, moderate or low dose) serum prepared on 6-week-old Sprague Dawley rats by serum pharmacologic method were respectively added into cultured OGC cell-line after treatment with testosterone propionat (TP). Then the cells were incubated for 48 hrs. The quantity of 3 H-thymidine incorporated DNA was determined with a scintillation counter; the cell cycle distribution and proliferation index (PI) were estimated with flow cytometer (FCM); and the contents of estrodial and progestogen in culture fluid and intracellular cAMP were detected by radioimmuoassay.

RESULTSExcessive androgen obviously inhibited the proliferation and secretion of OGC, and reduced the intracellular cAMP. BTR contained serum could improve above-mentioned inhibition of androgen to promote the cell proliferation and secretion of E2 and P in dose-dependant manner.

CONCLUSIONBTR could alleviate the inhibitory action of excessive androgen on OGC, the ovarian function may be adjusted by BTR through promoting follicular development, increasing the levels of estrogen and progestogen, improving the ratio of estrogen/androgen.

Animals ; Cell Proliferation ; drug effects ; Cells, Cultured ; Culture Media ; pharmacology ; Drugs, Chinese Herbal ; pharmacology ; Estradiol ; secretion ; Female ; Flow Cytometry ; Granulosa Cells ; cytology ; drug effects ; secretion ; Ovary ; cytology ; Rats ; Rats, Sprague-Dawley ; Serum ; Testosterone ; pharmacology

OBJECTIVETo study the anti-tumor effect and immune function modulation of Xiaoliu decoction (XLD) and to explore the possible mechanism.

METHODSS180 tumor bearing mice were treated with large or small dose of XLD and cyclophosphamide (CTX) to observe the tumor suppressive rate, lymphocyte transformation rate, natural killer cell (NK) activity, tumor necrosis factor alpha (TNFalpha) level in the mice, as well as the tumor cell apoptosis in vitro.

RESULTSThe tumor suppressive rate of XLD on tumor bearing mice was over 30%. XLD showed obvious trend in enhancing the tumor suppressing rate of chemotherapy, could increase the lymphocyte transformation rate, NK activity, and TNFalpha level. It also could accelerate the tumor cell apoptosis in vitro.

CONCLUSIONXLD has significant anti-tumor effect, it could elevate the immune function or organism and alleviate the chemotherapy induced lowering of immune function.

Adjuvants, Immunologic ; pharmacology ; therapeutic use ; Animals ; Antineoplastic Agents, Phytogenic ; pharmacology ; therapeutic use ; Cyclophosphamide ; adverse effects ; therapeutic use ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Female ; Killer Cells, Natural ; immunology ; Lymphocyte Activation ; drug effects ; Male ; Mice ; Phytotherapy ; Sarcoma 180 ; drug therapy ; immunology ; Tumor Necrosis Factor-alpha ; analysis

Mini health technology assessment (Mini-HTA) was developed from traditional HTA,based on the hospital needs.It is a very important decision making method and reference tool for the hospital policy makers.Currently there is no report of using Mini-HTA ease on introducing new equipment in China.Present paper introduces the Mini-HTA to provide reference for others hospitals in China.

Objective To identify the polyreactivity of a monoclonal natural anti-keratin autoantibody 3B4 and to analyze its possible molecular me chanism.Methods enzyme-linked immunosorbent assay (ELISA)and immunohistoche mistry were applied to test the binding reactivity of 3B4 against different anti gens and tissues.The variable region genes and their amino acid composition wer e sequenced.Results 3B4 could reacted with a range of antigens and tissues,i n addition to keratin and skin.The variable region genes of its light chain and heavy chain showed high homology with germline genes VK1 am4 and VH1 J558.42.H CDR3 region,which mainly composed of short side chain amino acids(from 294 to 324 nucleotides around the heavy chain),was the only motif that differs from ot her highly homologous immunoglobulin genes.Conclusions The monoclonal natural anti-keratin autoantibody 3B4,with its variable region genes highly homologo us to germline genes,is highly polyreactive.The flexibility of HCDR3 may contr ibute to the polyreactivity.

Objective To explore the cosmetic effect of oncoplastic volume displacement in breastconserving surgery.Methods 43 cases of breast cancer in stages I and Ⅱ with pathological diagnosis by core needle biopsy were operated by extended resection of tumor combined with axillary lymph nodes dissection.Among the patients,wound cavity was repaired by direct skin suture without suture of mammary gland deletion in 22 cases,and another 21 cases were repaired by oncoplastic volume displacement technique in which skin suture followed by freeing remaining breast tissue flap and translocation to repair the mammary defect.All of them were treated with three-dimension conformal radiotherapy,standard chemotherapy and endocrine therapy after operation.3 years after partial removal of breast,a retrospective historical control study had been fulfilled to analyze breast cosmetic results between two groups.Results In direct suture group,the cosmetology score was 4.3 points and fine rate was 40.9% (9/22).In breast reconstruction group.the score was 4.8 points and fine rate was 71.4% (15/21).The breast cosmetic effect in breast reconstruction group was significantly better than that in direct suture group (P<0.01).Conclusion It will get better breast cosmetic results after operation to utilize oncoplastic volume displacement suture technique in breast-conserving surgery.

Objective To observe the in vitro anti-Coxsackievirus B3m (CVB3m) effect of sophocarpine(SC) extracted from Sophora flavescens, a traditional Chinese herb. Methods HeLa cells were cultured and the micro-dose cytopathic effect (CPE) assays were applied to detect the toxicity of SC. CPE-inhibitory assays were used to observe the in vitro anti-CVB3m effect of SC. MTT and crystal assays were introduced to examine the anti-CVB3m effect of SC. HeLa cells were infected with CVB3m and added with SC in different concentrations 15 h later.The viability and number of survival of HeLa cells were determined by MTT and crystal violet assays, respectively. Results No toxicity was found on HeLa cells by SC with concentrations 100 ?g/mL, SC could accelerate and aggravate the CPE. SC could protect the CVB3m-infected HeLa cells with concentrations from 1.56 to 25 ?g/mL, and the viability and cell number measured by MTT and crystal violet assay in the SC-handled cells were higher and bigger than those in the virus infected ones. However, the inhibitory effect of virus was exacerbated with higher concentrations (50 and 100 ?g/mL), and the cell number and viability of the SC-handled cells were smaller and lower than those of the infected ones. Conclusion SC with a proper concentration has the in vitro anti-CVB3m effect and may protect HeLa cells from CVB3m infection.

Objective To investigate the relationship between alteration of gene in cyclic adenosine monophosphate(cAMP) signal transduction system in rats after myocardial damage and changes of cardiac function and ventricular remodeling. Methods Twenty eight male Wistar rats weighing 220 g to 250 g were randomly divided into three groups: acute myocardial damage group(AMD, n = 10), chronic myocardial damage group (CMD,n = 9 ) and sham-operation group (control, n = 9). Animal model of acute myocardial damage was established by ligation of rats left coronary artery in the AMD and the CMD groups. Rats in control group were treated similarly, except that the coronary suture was not tied. Hemodynamics and echocardiography were measured before rats were sacrificed 24 hours after operation in control and AMD groups but those in CMD groups were sacrificed 8 weeks later. Cadiocyte apoptosis were estimated by TUNEL method, cAMP levels in heart were tested by radioimmunity and the mRNA expressions for inducible cAMP early repressor (ICER), cAMP response element binding protein (CREB), phosphodiesterase 3A (PDE3A) and bcl-2 were assayed by real-time quantitative reverse transcriptase polymerase chain reaction (RT-PCR). Results The difference of left ventricular end diastolic diameter (LVEDD), left ventricular end diastolic pressure(LVEDP), maximal rising and falling rate of ventricular pressure,left ventricular systolic pressure (LVSP), eject fraction (EF) and fraction shortening (FS) were statistically significant among the three groups(F = 285.9, 196.8, 83.2, 80.4, 54.9, 196.6, 95.2, all P ＜ 0.01). LVEDD[(7.03 ±0.28), (8.20 ± 0.27)mm] and LVEDP[(11.19 ± 2.89), (19.76 ± 3.34)mmHg] in AMD and CMD groups were significantly increased, compared with those in control group[ (5.05 ± 0.30)mm, (- 5.62 ± 3.01 )mmHg, all P ＜0.01 ]. While maximal rising rate[ (2964 ± 449), (2214 ± 434)mmHg/s] and falling rate[(- 2617 ± 441),(- 1891± 424)mmHg/s] of left ventricular pressure, LVSP[ (94.19 ± 4.03), (85.85 ± 6.39)mmHg], EF[ (41.6 ±5.9)%, (35.9 ± 4.1 )%] and FS[ (36.9 ± 4.6)%, (23.1 ± 4.9)%] of left ventricular in the two groups were lower than those in control[(4759 ± 406)mmHg/s, (- 4327 ± 388)mmHg/s, (116.29 ± 8.25)mmHg, (80.9 ± 5.6)%,(53.1 ± 4.3)%, all P ＜ 0.01 ]. These changes in CMD group were more significant than those in AMD groups(P ＜0.05 or P ＜ 0.01 ). The difference of apoptotic index, cAMP and expression of ICER, CREB, PDE3A mRNA and bcl-2 mRNA were statistically significant among the three groups(F= 172.5, 141.0, 540.8, 246.8, 165.1, 563.9,all P＜ 0.01 ). Apoptotic index[ (32.8 ± 4.2)‰, (18.4 ± 3.9)‰] and cAMP in heart[ (9.95 ± 0.30), (5.60 ± 0.25)nmol/kg] in AMD and CMD groups were increased compared to control group[ (3.9 ± 1.7)‰, (2.48 ± 0.29)nmol/kg,all P ＜ 0.01 ], and those in CMD group were lower than in AMD group(all P ＜ 0.01 ). Expression of ICER mRNA (1.434 ± 0.093, 0.942 ± 0.076) and CREB mRNA(5.70 ± 0.50, 2.64 ± 0.51) in AMD and CMD groups were higher, and expression of PDE3A mRNA(48.98 ± 8.14, 16.68 ± 8.46) were lower than those in control group (0.154 ± 0.063, 1.08 ± 0.35, 105.94 ± 12.61, all P ＜ 0.01 ). The three genes in CMD group were fewer than those in AMD group(all P ＜ 0.01 ). bcl-2 mRNA was up regulated in AMD group(4.55 ± 0.27) and was down regulated in CMD group(0.35 ± 0.15) compared to control(2.18 ± 0.30, all P＜ 0.01). Conclusions There is PDE3A-ICER positive-feedback loop leading to myocyte apoptosis and heart failure after myocardial damage. The downregulation of PDE3A mRNA observed in chronic myocardial damage may play a causative role in the progression of ventricular remodeling and heart failure, in part, by inducing ICER mRNA and promoting cardiac myocyte dysfunction.