Objective To get a new approach for conserving the germplasm of Siraitia grosvenorii Methods Shoots, 0.8—1 cm in length, excised from test-tube plantlets which were subcultured 15 d, were precultured. Then its shoot-tips about 2—3 mm in length were dissected and loaded with 60% PVS2 at 25 ℃, and dehydrated with 100% PVS2 at 0 ℃, changed for fresh 100% PVS2 prior to directly plunging into liquid nitrogen. After cryopreservation for 24 h, the shoot tips were thawed, rinsed in MS+1.2 mol/L sucrose medium, blotted with filter papers, then plated on the MS + 1.0 mg/L 6-BA+0.05 mg/L NAA + 0.1 mg/LGA3 + 0.8% agar + 45 g/L sucrose medium at 25 ℃ for 7 d in dark prior to exposure to the light. The root medium for regeneration plantlets was 1/2 MS + 0.2 mg/L NAA + 30 g/L sucrose. Results Shoots were precultured for 3 d on MS + 0.7 mol/L sucrose medium, then its shoot-tips loaded for 40 min, dehydrated for 50 min. After cryopreservation, the shoot tips were rapidly thawed in water at 40 ℃, then rinsed for 40 min. The survival rate of shoot-tips plated on the recovering medium in one week was 100%, and regeneration rate after 30 d was 78.33%, which was the highest. The regeneration plantlets inoculated on root medium were reconstructed integrating plantlets. Conclusion The method of vitrification to cryopreserve the germplasm of S. grosvenorii is a simple way with high survival rate and normal regeneration plants.

Objective To study the effects of estrogens ( 17α-ethynylestradiol and 17β-estradiol ) on the ATP-sensitive potassium channel ( KATP ) activities in the enzymatically ( collagenase and protease ) isolated ICR mouse ventricular cardiomyocytes using inside-out and cell-attached patch clamp techniques .Methods In the inside-out patch configurations, the two estrogens reduced the K ATP channel activities in a dose-dependent manner at -60 mV holding potential ( HP) .Results The half-maximal inhibitory concentration ( IC50 ) of the 17α-esthynylestradiol and the 17β-estradiol were 0.3μmol/L and 0.1 nmol/L, respectively.However, these agents had no effects on the pinacidil-and DNP-induced KATP channel activities in the cell-attached patch configuration.However, the inhibitory effect of KATP channel activities by the 17-estrogens or the anti-estrogens in the excised inside-out patch configuration were abolished by 1 pmol/L PDBu.Conclusions The results of this study demonstrate that some estrogens decrease the K ATP channel activity acting on the inside of the isolated cardiomyocytes , and play a protective role to cardiomyocytes .

Objective To develop a sensitive and stable substrate for enzyme‐linked immunosorbent assay(ELISA)and carry out methodology evaluation tests on it .Methods A kind of substrate with tetramethyl benzidine and urea peroxide as its main compo‐nents were made up ,its sensitivity ,precision and storage stability were evaluated and compared with other three kinds of substrate . Results This substrate was more sensitive than two of the three commercial substrates .The variable coefficient of precision test was 3 .5% .The optical densities(OD) of the new substrate after stored one day ,seven days ,one month ,six months ,twelve months in 4 ℃ environment were 2 .268 、2 .403 、2 .358 、2 .278 、2 .330 respectively ,the standard deviation was 0 .056 185 ,the variable coeffi‐cient was 2 .4% .Conclusion The substrate proposed in this paper displays good sensitivity ,precision and stability .The preparation method is simple ,reproducible .This substrate not only satisfies the requirements of laboratory research ,but also meets with the de‐mands for commercial kit development due to its function as an important component of test kits .It is an economical and effective choice for both laboratories and research transformation .

Objective To study the prevalence of symptomatic osteoarthritis (OA) in rural areas of Shanxi,China.Methods All the residents above 16 years old from the villages chosen by multi-stage stratified cluster sampling in Yangcheng County and Pianguan County in Shanxi Province were investigated with COPCORD procedure of WHO.All the respondents signed the informed consent forms.All statistical analysis were carried out using SAS 9.2 and SPSS standard version 13.0.Chi-square test was used for statistical analysis.P value ＜0.05 was regarded as statistically significant.Results The investigation in these two counties covered 7 126 residents,including 1 734 symptomatic OA patients,with a prevalence rate of 24.3％,where knee OA had the highest prevalence rate [13.8％(983/7 126)],other figures were as follows:lumbar vertebra,neck,hand,shoulder,elbow,ankle,hip,wrist,thoracic vertebra and foot.There were statistical differences in the prevalence of knee,ankle,neck,shoulder and hand between the two genders (P＜0.05).Conclusion The prevalence of symptomatic OA in rural areas of Shanxi,China is relatively high,but the risk factors still need to be further studied and proved.

OBJECTIVE To analyze the values of drug-resistant Pseudomonas aeruginosa pneumonia in the older patients using thymosin ?1 combined with Sulperazone.METHODS The thirty five older patients with drug-resistant P.aeruginosa pneumonia were divided into two groups,one group was treated with thymosin ?1 combined with Sulperazone,the other group was treated with Sulperazone alone.We observed the changes in clinical symptom of the patients,results in culture of sputum,routine blood test,C-reactive protein(C-RP),albumin,pre-albumin,and T cell subset before and after drug use.RESULTS The clinical symptoms of the patients were improved,days of disappeared drug resistance of P.aeruginosa,reconverted normally days of routine blood test,decreased concentration of CRP,albumin,and pre-albumin reconverted to normalize,increased percent of CD4+,decreased percent of CD8+,CD4+ /CD8+ were more prominently reconverted to normalize in thymosin ?1 combined with Sulperazone treated group than in Sulperazone alone group.CONCLUSIONS Thymosin ?1 can improve immunological function,inflammation condition and protein metabolism.Its application with Sulperazone has a coordinated effect on drug-resistant P.aeruginosa pneumonia in the older patients.

Objective To investigate the expression of vascular endothelial growth factor C(VEGF-C) and vascular endothelial growth factor receptor-3(VEGFR-3) in rat colorectal cancer,and to provide an experimental material for the role of VEGF-C and its receptor VEGFR-3 in cancer progression and metastasis via lymphatics. Methods The expressions of VEGF-C and its receptor VEGFR-3 in colorectal cancer induced by MNNG were observed by immunohistochemistry staining. Results There was no expression of VEGF-C in normal colorectal tissues,but VEGFR-3 expressed in the lymphatic endothelium.The protein of VEGF-C mainly expressed in the cancer cells and the positive rates were 75% and 100% respectively in the early and the mid-terminal stages of cancer.The positive expression of VEGF-C in the early stage of cancer was higher than that in mid-terminal stage(P

Objective To explore the influence of EGFR gene interfering on the radiosensitivity of esophageal squamous cells.Methods Three kinds of siRNAs including three random sequences of positive EGFR siRNA (EGFR siRNA1、EGFR siRNA2、EGFR siRNA3),random sequence of negative EGFR siRNA,and blank control were transfected into Eca109 cells by lipofectamine.Cell proliferation was detected by Cell Counting Kit-8 assay.Protein and mRNA expressions of EGFR were detected by Western blot and RT-PCR,respectively.The ability of cell clone formation was used to evaluate the combination effect of X-rays and EGFR siRNA on the radiosensitivity.Results The positive expression rate of the EGFR mRNA in the Eca109 ceils transfected with EGFR siRNA1,EGFR siRNA2,EGFR siRNA3 was 26.74％,9.52％,4.61％,respectively,which was significantly lower than 42.44％ in the control cells transfected with blank siRNA (F =112.11,P ＜ 0.01).Meanwhile,the EGFR protein expression was reduced by 72.84％,53.01％ and 56.21％ after interfering of siRNA1,siRNA2,and siRNA3,respectively.CCK8 assay showed that the proliferation of Eca109 cells was decreased by 28.2％ since the siRNA interference.Moreover,the D0,Dq and SF2 of the combined treatment group were lower than those of irradiation alone group and the sensitization enhancement ratio was 1.50.Conclusions EGFR siRNA can effectively inhibit EGFR gene expression and enhance the radiosensitivity of Eca109 cells.

Objective To study the clinical characteristics, diagnostic methods and therapeutic efficacy of pneumocystis pneumonia (PCP) in patients with acquired immunodeficiency syndrome (AIDS). Methods Sixty-nine AIDS cases of PCP were diagnosed according to the criteria of USA Centers for Disease Control and Prevention revised in 1993. The clinical symptoms and signs of the patients were observed. The peripheral blood lymphocyte counts, blood gas analysis and bronchoalveolar lavage fluid (BALF) were checked and transbronchoscopic lung biopsy was performed. Results All studied patients were in the late stage of AIDS. The main clinical manifestations included fever (100.0%), cough (97.1%), and dyspnea (92.80%). Pulmonary rales could be heard in 42 cases (60.9% ). Peripheral CD4+ T lymphocyte counts ranged from 1 × 106 -88 × 106/L. Fifty-two cases (75.4% ) had low arterial partial pressure of oxygen value of less than 10.7 kPa (1 kPa = 7.5 mm Hg). Sixty-one cases (88.4 %) had elevated serum lactate dehydrogenase (LDH) level. Bilateral diffused interstitial change (46.4%) and ground-glass shadow (29.0%) were the most common abnormal chest radiological findings. Pneumocystis organisms were detected in the BALF from 2 patients and in the transbronchial biopsy (TBB) tissue from 35 patients. All patients were treated with compound sulfamethoxazole. Thirty-three were treated with corticosteroid simultaneously and 27 were assisted with mechanical ventilation. Fifty patients recovered or got improved, eleven died, and eight left hospital because of deteriorated condition. Conclusions When an AIDS patient represents with fever, cough, dyspnea, hypoxemia, elevated serum I.DH level, CD4+ T lymphocyte count below 100 × 106/L, and interstitial pneumonia or ground-glass shadow in chest images, the diagnosis of PCP could be made presumptively. It is difficult to make a nosogenic diagnosis of PCP, but TBB considerably increases the positive rate of pneumocystis. Compound sulfamethoxazole is recommended as the first selected drug. In severe cases, corticosteroid and assisted mechanical ventilation combined with compound sulfamethoxazole could remarkably improve the prognosis of PCP.

An analytical method of ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC/PDA-QDa) for the qualitative and quantitative analysis of Δ9-tetrahydrocannabinol (Δ9-THC), cannabidiol (CBD), and cannabinol (CBN) in confiscated cannabis was developed.The seized cannabis was extracted in methanol by sonication.The binary mobile phase consisted of methanol (containing 0.1% formic acid) and water.After centrifugation, the supernatant was separated on Waters UPLC BEH C18 column (50 mm×2.1 mm, 1.7 μm) with isocratic elution at a flow rate of 0.2 mL/min.The three cannabinoids were analyzed by photodiode array (PDA) detector at 220 nm and confirmed by mass spectrometer QDa.The correlation coefficient of standard curve for the three cannabinoids in linearity range was not less than 0.999, as well as the recoveries were 82%-102% with the relative standard deviations (RSD) of 0.36%-4.12% at three spiked levels.The method is specific, easy, quick and suitable for confirmation of the cannabinoids in seized cannabis.Cannabis plants in different areas were classified by their chemical phenotype as drug-type or fiber-type plants, taking into account the phenotypic index Δ9-THC, (Δ9-THC+CBN)/CBD, or the Δ9-THC/CBD and the (Δ9-THC+CBN)/CBD ratios.The analysis of the original composition of plant material is necessary for the detection and the quality control of cannabis plants.

Objective To study the pathogens and drug resistance profiles of pulmonary infection in patients with AIDS. Methods The pathogens and their drug susceptibility of pulmonary infection diagnosed by fibrobronchescopy-induced brunchoalveolar lavage fluid (BAI.F) culture and/or transbronchial biopsy in 116 AIDS cases were analyzed. Results Monopathogenic infection in lungs were detected in 18 cases(15.5%) and mixed infection in 98 cases ( 84.5%). Of the 116 cases, bacteria were present in 91 patients, fungi in 62, tubercle bacillus in 49, pneumocystis jiroveci in 29, and cytomegalovirus in 11.Ninety-five bacterial strains were isolated from BALF, mainly including Streptococci (34), coagulase negative Staphylococcus (20), Klebsiella pneumoniae (10) and Escherichia (7). The isolated bacteria were resistant to β-lactam, macrolides, quinolones and aminoglycosides, of which were 14 methicillin-resistant Streptococci (MRS) strains and 12 extended spectrum β-lactamases (ESBL) strains. Sixty-eight fungal strains were isolated, including 36 Candida mycodermas, 19 Penicilliums, 6 Aspergilli and 5 Mold fungi;they were sensitive to amphotericin B but resistant to fluconazol (5.6% -50. 0% ) and itraconazole( 10. 5%-60. 0% ). Conclusion Pneumonia in AIDS patients are usually caused by multiple pathogens,predominantly consisting of multiresistant bacteria and fungi. Therefore, antibiotics should be rationally chosen according to drug susceptibility test.