The viability and life span of pollen were evaluated by TTC (2,3,5-triphenyl tetrazlium chloride) and the peroxidase solution, the stigma receptivity were estimated by benzidine-H2O2 method and the fruiting characteristics were investigated. The results showed that (1) Anoectochilus roxburghii and A. formosanus appeared the same up-and-down trend of the pollen viability, increased and then decreased. The storage temperature and storage time had significant impact on the pollen viability. With the extension of storage time, the pollen activity decreased. 4 degrees C refrigerator storage may be extended the pollen vitality. (2) The stigma had receptivity in 1st day and reached the highest level in the 4th day after blooming. A. roxburghii lost receptivity in the 8th day while A. formosanus lost receptivity in the 10th day after blooming. (3) The different pollination had significant impact on seed setting rate. The seed setting rate of artificial cross-pollination was higher than that of the artificial self-pollination. Collecting pollen in the 3rd day and carrying out artificial cross-pollination in the 4th day after blooming can significantly improve seed setting rate. The results provided technical assurance for A. roxburghii and A. formosanus breeding of new varieties and seed breeding.
Cell Survival ; China ; Flowers ; physiology ; Fruit ; growth & development ; Orchidaceae ; growth & development ; physiology ; Pollen ; growth & development ; Reproduction ; Temperature

This work was mainly studied the effects of the four alkaloids from Coptidis Rhizoma on the mouse peritoneal macrophages in vitro and preliminarily discussed the regulating mechanisms. The effect of alkaloids from Coptidis Rhizoma on the vitality of macrophages was measured by the MTT assay. The effect of alkaloids on the phagocytosis of macrophages was determined by neutral red trial and respiratory burst activity was tested by NBT. The expressions of respiratory-burst-associated genes influenced by alkaloids were detected by qRT-PCR. The conformation change of membrane protein in macrophages by the impact of alkaloids was studied by fluorospectro-photometer. Results showed that the four alkaloids from Coptidis Rhizoma could increase the phagocytosis of macrophages in different level and berberine had the best effect. Berberine, coptisine and palmatine had up-regulation effects on respiratory burst activity of mouse peritoneal macrophages stimulated by PMA and regulatory activity on the mRNA expression of PKC, p40phox or p47phox, whereas the epiberberine had no significant influence on respiratory burst. Moreover, alkaloids from Coptidis Rhizoma could change the conformation of membrane protein and the berberine showed the strongest activity. The results suggested that the four alkaloids from Coptidis Rhizoma might activate macrophages through changing the conformation of membrane protein of macrophages and then enhanced the phagocytosis and respiratory burst activity of macrophages. Furthermore, the regulatory mechanism of alkaloids on the respiratory burst activity of macrophages may be also related to the expression level of PKC, p40phox and p47phox.
Alkaloids ; pharmacology ; Animals ; Cells, Cultured ; Coptis ; chemistry ; Drugs, Chinese Herbal ; pharmacology ; Female ; Gene Expression ; drug effects ; Macrophages, Peritoneal ; drug effects ; Mice ; Phosphoproteins ; genetics ; metabolism ; Protein Kinase C ; genetics ; metabolism ; Rhizome ; chemistry

Objective To establish qualitative and quantitative analysis methods for identification and determination of multi-constituent of Chanfukang granules (CFKG) using high performance liquid chromatography-time-of-flight mass spectrometry (HPLC-TOF-MS) and high performance liquid chromatography-triple quadrupole mass spectrometry (HPLC-MS/MS),and a preliminary pharmacokinetic study was carried out.Methods An Agilent ZORBAX Eclipse Plus C1s column (100 mm × 3.0 mm,1.8 μm) and a Waters XBridge(R) BEH C18 column (150 mm × 4.6 mm,2.5 μm) were used with 0.1％ formic acid aqueous solution-acetonitrile as mobile phases by gradient elution.The compounds were detected by electrospray ion source in both positive and negative mode with multiple reaction monitoring (MRM) mode.SD rats were ig with CFKG (0.5,5 g/kg).Blood samples of 0.2 mL were taken from jugular vein at different time points and plasma components and contents were determined by HPLC-MS/MS.Results Twenty-four constituents were identified by HPLC-TOF-MS qualitative analysis and thirteen constituents were quantitatively detected by HPLC-MS/MS.The established HPLC-MS/MS method had a good separation,and all the legal verification met the requirements.The content of stachydrine,Leonurine,and astragaloside Ⅳ were high,which may be the main active ingredient The pharmacokinetic results showed that,only when the dose was 5 mg/kg,stachydrine and Leonurine can detected.Elimination of stachydrine was slow and elimination of Leonurine was fast.Conclusion A rapid and efficient method for studying the chemical constituents was established,which could provide reference for the quality control of Chanfukang granules.

OBJECTIVETo investigate the association of SOX9 expression and clinicopathologic factors and prognosis of gastric cancer.

METHODSA retrospective cohort study including 112 gastric cancer patients admitted to the Zhejiang Provincial People's Hospital from 2004 to 2006 was performed. Immunohistochemical analysis was used to evaluate the expression of SOX9 in the 112 specimens of gastric cancer tissues and 70 non-cancerous tissues adjacent to the tumor.

RESULTSLow expression of SOX9 was seen in 5(7.1%) tissues out of 70 non-cancerous tissues adjacent to the tumor. A total of 94(83.9%) patients had varying expression of SOX9, of whom 51(45.4%) had overexpression. Univariate analysis demonstrated that the expression of SOX9 was significantly associated with Lauren classification (P<0.05), tumor invasion(P<0.01), lymph node metastasis(P<0.05), distant metastasis(P<0.05) and tumor stage(P<0.05), however there was no significant association between SOX9 expression and sex, age, histological type, histology differentiation or tumor size. Kaplan-Meier analysis showed that the 5-year survival rate of patients with SOX9 over-expression was significantly lower than that of patients with low expression(29.4% vs. 49.2%, P=0.031). Multivariate Cox regression analysis showed that histology differentiation(P=0.046), tumor invasion(P=0.001), and distant metastasis(P<0.01) were independent prognostic factors for gastric cancer, however the over-expression of SOX9 was not significant(P=0.948).

CONCLUSIONSThe expression SOX9 is associated with the growth, invasion, and metastasis of gastric cancer, as well as the prognosis. However, SOX9 expression is not an independent factor for the prognosis in patients with gastric cancer.

Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Kaplan-Meier Estimate ; Male ; Middle Aged ; Prognosis ; Retrospective Studies ; SOX9 Transcription Factor ; metabolism ; Stomach Neoplasms ; metabolism ; pathology

Objective To obtain guiding recommendations for the development of fertility preservation in female patients of reproductive age,so as to promote the scientific,standardized and orderly implementation of fertility preservation work.Methods A previous questionnaire survey was used to determine the preliminary recommendations,a modified Delphi method was used to invite 18 domestic experts of fertility preservation to conduct two rounds of expert consultation.Then we adjusted and summarized recommendations according to the results of the consultation.Results Finally,14 guiding recommendations for the development of fertility preservation in female patients of reproductive age were obtained according to the results of the Delphi consultation,including 5 recommendations at technical level,5 recommendations at political level and 4 recommendations at educational level.At technical level,it was proposed to establish a standardized operation process;at political level,it was proposed to improve the relevant legislation of fertility preservation in China to protect the safety and interests of patients;at educational level,it was suggested to take various forms to promote the mass dissemination of fertility preservation knowledge.Conclusion Guiding recommendations for fertility preservation in female patients of reproductive age can provide detailed and comprehensive guidance for promoting fertility preservation in female patients of reproductive age in Shanghai,and provide reference for carrying out fertility preservation in other regions.

OBJECTIVETo study the incidence and risk factors of HIV and syphilis seroconversion among men who have sex with men (MSM) in Beijing.

METHODSA total of 550 MSM were recruited on the basis of community and followed up after 6 and 12 months in Beijing. Each subject was investigated by only one investigator at one time to collect information on demographics and behaviors. Blood samples were collected to test HIV and syphilis seroconversion. ELISA was used for screening test, west blotting (WB) and Particle agglutination were used for confirmatory test.

RESULTSA total of 550 MSM investigated, among which 4.5% (25/550) were HIV-positive and 29.3% (161/550) were syphilis-positive. For 525 HIV-negative MSM, 87.0% (457/525) retained during the 12-month investigation. Seroincidence for HIV and syphilis were 3.37/100 person-years (95%CI = 1.66 - 5.08) and 9.32/100 person-years (95%CI = 5.87 - 12.77) respectively. HIV seroconversions for those who performed and did not perform rectal douching after homosexual anal intercourse in the past 3 months were 7.11/100 and 0.76/100 person-years respectively. Multivariate Cox regression analysis revealed that rectal douching after homosexual anal intercourse in the past 3 months (HR = 9.23, 95%CI = 2.08 - 40.88) was significantly associated with HIV seroconversion. Syphilis seroconversions for those who met male sex partners in parks, public washrooms or bathhouses in the past 3 months were 41.77/100 and 7.97/100 person-years respectively. Syphilis seroconversions for those who performed and did not perform rectal douching after homosexual anal intercourse in the past 3 months were 16.17/100 and 4.92/100 person-years respectively. In the past 3 months, meeting male sex partners in parks, public washrooms or bathhouses (HR = 4.67, 95%CI = 1.77 - 12.34) and performing rectal douching after homosexual anal intercourse (HR = 3.09, 95%CI = 1.40 - 6.83) were significantly associated with syphilis seroconversion.

CONCLUSIONThe seroconversions of HIV and syphilis during the follow-up visits in this MSM cohort study in Beijing were very serious, and that the associated factors for seroconversions were rectal douching after homosexual anal intercourse and meeting male sex partners in parks, public washrooms or bathhouses.

Adolescent ; Adult ; Antibodies, Bacterial ; blood ; China ; epidemiology ; HIV ; immunology ; HIV Antibodies ; blood ; HIV Infections ; blood ; epidemiology ; HIV Seropositivity ; blood ; epidemiology ; Homosexuality, Male ; Humans ; Incidence ; Male ; Risk Factors ; Sexual Behavior ; Syphilis ; blood ; epidemiology ; Treponema pallidum ; immunology ; Young Adult

OBJECTIVETo explore the effects of the magnetic c-erbB-2 antisense probe of different concentrations on the morphology and expression of SK-Br-3 cancer cells in vitro.

METHODSBreast cancer SK-Br-3 cells were transfected for 24 h by antisense probe at an iron concentration of 5, 10, 25, 50, and 100 mg/L, respectively. The distribution and content of iron particles in SK-Br-3 cells was determined by Prussian blue staining, electron microscopy, and atomic absorption spectrometry. Cell viability was observed by trypan-blue exclusion and CCK-8 test. The protein expression of c-erbB-2 was assessed by the Western blot analysis. The changes of the signal strength were considered by magnetic resonance imaging (MRI).

RESULTSc-erbB-2 antisense probe was uptake by SK-Br-3 cells in a concentration-dependence manner within a certain range (5, 10, and the Medicine Scientific Research Project of Chongqing Health Bureau (062025)25 mg/L). When the probe concentration was 25 mg/L, iron content in cells was (18.38±0.28) pg, the cell vitality, survival, and c-erbB-2 protein expression were reduced significantly (all P<0.05), and the T2 value was lower significantly (P<0.05). However, the results of 50 mg/L or 100 mg/L group showed no significant difference with the 25 mg/L group (P>0.05).

CONCLUSIONThe magnetic c-erbB-2 antisense probe can effectively transfect and specifically inhibit the expression of SK-Br-3 cell lines at the iron concentration of 25 mg/L.

Antisense Elements (Genetics) ; genetics ; Apoptosis ; Breast Neoplasms ; metabolism ; pathology ; Cell Line, Tumor ; Cell Proliferation ; Humans ; Magnetics ; Receptor, ErbB-2 ; genetics ; metabolism ; Transfection

Objective To observe the therapeutic effect of Lai's acupuncture therapy combined with Chinese herbal decoction for the treatment of infertility with ovulatory dysfunction, and to summarize the experience of Professor Lai Xin-Sheng in treating infertility. Methods Ninety-six infertility patients with ovulatory dysfunction were randomized into treatment group and control group, 48 cases in each group. The treatment group was treated with Lai's Tongyuan acupuncture therapy for regulating Governor Vessel to tranquilize mind, inducing meridian Qi to original Qi, and was also given oral use of Chinese herbal decoction of Lai's Yiyun Recipe for promoting gestation. The control group was treated with Chinese herbal decoction alone. The treatment began exactly after the completion of menstruation period and lasted 15 days for one treatment course, covering 3 courses. And then the patients of the two groups were followed-up for 3 months. Therapeutic effect and pregnancy rate were evaluated after treatment, serum levels of estradiol and progesterone were examined, and abdominal ultrasonography was used for monitoring the maximum diameter of the dominant follicles and endometrial thickness before and after treatment. Results (1) Three cases from the treatment group and 5 cases from the control group were dropped out, and the final case number in the treatment group and control group was 45, 43 respectively. (2) The total efficiency of the treatment group was 88.9％, and that of the control group was 81.4％. The effect of the treatment group was better than that of the control group (P < 0.05). (3) The pregnancy rate of the treatment group was 73.3％, higher than that of the control group (51.2％), the difference being significant (P < 0.05). (4) After treatment, the maximum diameter of the dominant follicles and endometrial thickness in the two groups were obviously improved(P < 0.05 compared with those before treatment), and the improvement in the treatment group was superior to that in the control group (P < 0.05). (5) After treatment, the serum levels of estradiol and progesterone in the two groups were obviously improved(P < 0.05 compared with those before treatment) , and the improvement in the treatment group was superior to that in the control group(P < 0.05). Conclusion The therapeutic effect of Lai's acupuncture therapy combined with Chinese herbal decoction is superior to that of Chinese herbal decoction alone, showing an obvious synergistic effect by shortening the course of treatment and enhancing pregnancy rate.

OBJECTIVETo study whether combined detection of the methylation status of vimentin, sFRP1, and HPP1 gene can increase the positive methylation rate in colorectal cancer.

METHODSTissue samples were collected from 90 patients with colorectal cancer, 60 patients with adenomatous polyp, and 20 healthy controls. DNA was extracted and the methylation status of vimentin, sFRP1, and HPP1 gene was detected by Methylation-specific PCR (MSP). The relationship between clinicopathologic features of colorectal cancer and gene methylation was analyzed.

RESULTSThe methylation rates of vimentin, sFRP1, and HPP1 were 66.7%, 68.9%, and 72.2% in colorectal cancer, 53.3%, 55.0%, and 50.0% in colorectal adenomas, and 0, 0, and 5.0% in healthy controls, respectively. The methylation of each of the three genes in colorectal cancer tissues was higher than colorectal adenomas and healthy controls(P<0.05). The diagnostic sensitivity by combining three methylation markers was 93.3% in colorectal cancer, 76.7% in colorectal adenomas, which was higher than the sensitivity using single gene testing(P<0.05). No significant associations existed between the methylation status of the three genes and clinical characteristics including sex, age, tumor location, lymph node metastases, distant metastasis, and TNM stage(P>0.05).

CONCLUSIONSDNA methylation levels of vimentin, sFRP1 and HPP1 are significantly higher in colorectal cancer tissue. Combined detection significantly improves the positive rate of methylation, and may be used as early diagnosis method for colorectal cancer.

Adult ; Aged ; Aged, 80 and over ; Ataxia Telangiectasia Mutated Proteins ; genetics ; Case-Control Studies ; Colorectal Neoplasms ; diagnosis ; genetics ; DNA Methylation ; Female ; Humans ; Male ; Membrane Proteins ; genetics ; Middle Aged ; Neoplasm Proteins ; genetics ; Promoter Regions, Genetic ; genetics ; Vimentin ; genetics

Biopsy ; Calcitriol ; therapeutic use ; Calcium ; therapeutic use ; Fanconi Syndrome ; diagnosis ; drug therapy ; Humans ; Immunosuppressive Agents ; therapeutic use ; Pilot Projects ; Prednisone ; therapeutic use