To clone the differential genes in antibiotic resistence Neisseria gonorrhoeae,the library that contains the fragments of differential genes between antibiotic resistance strain and standard reference strain of Neisseria gonorrhoeae was constructed which used suppression subtractive hybridization technique. Then the antibiotic resistance relational genes fragments were cloned and analyzed. Antibiotic resistance Neisseria gonorrhoeae subtractive library that has high subtractive efficiency was set up successfully. The amplified library contained 2500 positive clones. Sequence analysis was performed to find the fragments of antibiotic resistance relational genes. Five sequences were unknown previously. The fragments of antibiotic resistance genes may provide an important clue for studying the mechanism of occurrence and development of antibiotic resistance Neisseria gonorrhoeae.

BACKGROUND: Traditional solid bone can not receive satisfied effects in repairing irregular bone defects in oral maxillofacial surgery due to uneven distribution of cells and growth factors. Therefore, it is a research direction to prepare injectable tissue engineering bone.OBJECTIVE: To explore the effects of chitosan/β-tricalcium phosphate/recombinant human bone morphogenetic protein-2(CS/β-TCP/rhBMP-2) composite on mandibular defect repair.DESIGN, TIME AND SETTING: A randomized controlled animal experiment. The experiment was performed atthe animal laboratory, Medical College of Jinnan University from May 2008 to March 2009.MATERIALS: The injectable tissue engineering bone was prepared by using complex of liquid CS and solid β-TCP as scaffold materials, and combined with freeze-dried rhBMP-2.METHODS: Twenty-four New Zealand white rabbits were prepared for double sides mandibular defect models, and randomized into 4 groups: ①CS/p-TCP/rhBMP-2 group: 1 mL CS/β-TCP/rhBMP-2 complex was injected into the defects.②CS/β-TCP group:0.5 mL CS/β-TCP complex was injected into defects. ③Autograft bone group: repairing the defects with sclerotin of the iliac crest.④Blank control group: no implantation. MAIN OUTCOME MEASURES: At weeks 2, 4 and 8 after surgery, the material degradation and new bone formation were evaluated with, haematoxylin-eosin staining, and electron microscope; the bone mineral density was detected by dual energy X-ray absorptiometry (DXA) to determine bone formation rate and quality.RESULTS: ①Gross observation demonstrated that the size and thickness of osteotylus in CS/β-TCP/rhBMP-2 group was equivalent with the autograft group, which were greater than that of the other groups.②Histologicalobservation demonstrated that there were more bone matrixes in the CS/β-TCP/rhBMP-2 group and autograft group than that in the CS/β-TCP group and blank control group at each time points. ③Scanning electron microscope image suggested that at 8 weeks after operation, the bone bed and the materials in CS/β-TCP/rhBMP-2 group were connected with bone, and the gap was diminished. The degradation of the materials was so obvious that the complete structure of materials could not be found. ⑤DXA detection appealed that the bone density of each group was gradually increased with time prolonged. The quantities of bone density in CS/β-TCP/rhBMP-2 group in weeks 2, 4 and 8 were significantly higher than CS/β-TCP group and blankcontrol group (P < 0.05).CONCLUSION: ①CS/β-TCP/rhBMP-2 has good biocompatibility, degradability and the capacity of bone guidance and bone induction. ②CS/β-TCP can be served as a promising carrier for BMP-2, which is a potential degradable biological material for repairing bone defects.

Fluorescence enhancement reaction of flavoxate hydrochloride (FX) in strong alkali solution was studied, the mechanism of the reaction was investigated, and a novel fluorimetric method for analysis of FX in drug sample was established. FX has no intrinsic fluorescence, but it can slowly produce fluorescence in strong alkali solution. Heating can promote the fluorescence enhancement reaction. In 3D fluorescence spectra of the decomposition product of FX, two fluorescence peaks, located respectively at excitation wavelengths λex/ emission wavelength λem =223/410 nm, and 302/410 nm, were observed. Using quinine sulfate as a reference, fluorescence quantum yield of the decomposition product was measured to be 0.50. The structural characteriza- tion and spectral analysis of the decomposition product reveal that ester bond hydrolysis reaction of FX is firstly occurred during heating process, forming 3-methylflavone-8-carboxylic acid (MFA), then a cleavage reaction of the γ-pyrone ring of MFA occurred, producing α, β-unsaturated ketone. This product includes adjacent hydroxyl benzoic acid group in its molecule, which can form intramolecular hydrogen bond under alkaline condition, so that increase the conjugate degree and enhance the rigidity of the molecule, and thereby cause fluorescence enhancement. Based on this fluorescence enhancement reaction, a fluorimetric method was proposed for the determination of FX. A linear calibration curve covered the concentration range 0.020 3-0.487 µg · mL. The regression equation was I(F) = 23.9 + 5357.3 c, with correlation coefficient r = 0.999 7 (n = 8), detection limit D = 1.1 ng · mL(-1). The method was applied to the analysis of FX tablets, with a spiked recovery rate of 100.2%. The reliability of the method was verified by a UV-spectrophotometric method.
Alkalies ; Calibration ; Chemistry, Pharmaceutical ; Flavoxate ; analogs & derivatives ; chemistry ; Fluorescence ; Limit of Detection ; Reproducibility of Results ; Solutions ; Tablets

Objective To determine whether homocysteine(Hcy)and monosodium glutamate (MSG)could lead to animal model of hypertensive disorder complicating pregnancy and its mechanism. Methods Female adult Wistar rats were randomly divided into 4 groups:pregnant control group(PN), pregnant Hcy group(PH),pregnant glutamic acid group(PG)and pregnant Hcy and glutamie acid group (PHG).The rats of each group were injected with Hey 200 mg/kg or physiological saline every day intraperitoneally and with MSG or 0.9% saline every other day via Hcy injection from the 10th day to the 20th day of pregnancy.The blood pressure,urine protein,function of liver and kidney,weight of placenta, length and weight of fetus were all measured.The histological change of the pallium and the change of behavior of pregnant rats were also observed.Results(1)The blood pressure in PH[(107?8)mm Hg, 1 mm Hg=0.133 kPa],and PHG group [(109?10)mm Hg] after the treatment increased significantly compared with those in other groups from the 12 th day after pregnancy(P

Objective To investigate the protective effects of electroacupuncture on the endothelial tissues of microvessels in the basal ganglia in the rats with cerebral ischemia-reperfusion.Methods The MCAO model was established by using Longa's method.The immunohistochemistry SABC(strepto-avidin-biotin-peroxidase complex) method was employed to detect the expression of ICAM-1,P-selectin in the microvessel of rats'ipsilateral basal gan- glia.Results The number of positive ICAM-1 and P-selectin endothelial cells of model group were significantly in- creased,as compared to normal group and sham operated group(P

AIM: To investigate the condition necessary for high-level expression of basic fibroblast growth factor (hbFGF) in Escherichia coli, a dicistron system was constructed. METHODS: The phage T7 gene ?10 N-terminal sequence, present in the expression plasmid pET-3c, was incorporated into the first cistron, the coding region for bFGF was incorporated into the second, and both were under the control of a transcription promotor recognized by the bacteriophage T7 RNA polymerase. RESULTS: Recombinants were transformed into BL21(DE3) and induced by IPTG for expression, accumulation of up to 20% of the total cell protein of bFGF were obtained, showing bioactivity indistinguishable from standard protein. CONCLUSION: It suggested that such dicistron system was effective for enhancing hbFGF expression.

Objective To explore the effects of different vitamin A(VA) levels on thyroid cells apoptosis and its gene expression of mice taking excessive iodine. Methods Kunming mice were randomly divided into 6 groups according to body weight 3 weeks after born: normal control(NI) group, high iodine(HI) group, low vitamin (LVA) group, high iodine plus low vitamin A(HI+LVA) group, high iodine plus vitamin A1 (HI+VA1) group, high iodine plus vitamin A2(HI+VA2) group. The VA was given in food(4000,4000,0,0,8000,16 000 U/kg), and the iodine was given as potassium iodate in water (I-:50,3000,50,3000,3000,3000 μg/L). The apoptosis was tested using in situ end labehng(TUNEL) method. Reverse transcription polymerase chain reaction (RT-PCR) were used to measure the level of mRNA of apoptosis gene(Fas, FasL, Bcl-2) in tissues. Results Apoptotic index measured by TUNEL method was rising along with the mice age. Compared to NI group[(14.09±5.68)%], apoptotic index was significantly increased in HI[(20.91±9.57)%], HI+LVA[(20.29±9.90)%]and HI+VA2 [(19.51±8.25)%]groups in the three months(P < 0.05). Compared to NI group[(16.80±9.90)%], apoptotic index was significantly increased(P < 0.05) in HI[(23.22±8.58)%],LVA[(22.56±6.17)%],HI+LVA [(25.99±9.62)%],HI+VA1 [(21.65±7.74)%]groups in the six months. Compared with the NI group(Fas: 1.29±0.25,1.27±0.26; FasL: 1.60±0.13,1.65±0.13), the mRNA levels of Fas and FasL in HI group(Fas: 1.57±0.36,1.49±0.35; FasL: 1.85±0.46,1.84±0.32) were increased, but the differences were not remarkable(P > 0.05) in the three and six months. Compared with the HI group, the mRNA levels of Fas in HI+ VA1, HI+VA2(1.33±0.35, 1.30±0.26) groups were decreased to the level in NI group in the six months. The mRNA levels of Fas and FasL were not different (P > 0.05) between HI+LVA(I.60±0.27,1.88±0.46) and HI groups in the three months. The mRNA levels of Bcl-2 were not remarkably differences in the three months (1.05±0.19,0.96±0.33,0.95±0.26,1.18±0.27,1.10±0.19,0.98±0.36, all P > 0.05), and in the six months (1.35±0.28,1.60±0.25,1.48±0.18,1.71±0.26,1.66±0.29,1.56±0.35, all P > 0.05). Conclusions Excessive iodine can cause thyroid cells apoptosis in mice. Supplementation of suitable amount of VA can regulate the levels of the apoptosis-related genes expression, and partly antagonize the apoptosis caused by high iodine.

Objective To explore incidence of child obesity in Zhengzhou area and intervention measures.Methods In 2001, spot check was conducted on 5688 cases of high and primary school students ,including 2848 boys and 2840 girls at the age of 7-18.Child obesity was diagnosed by meeting reference BMI value published by Cole et al,conducted comprehensive treatment consisting of massage intervention along channel of traditional Chinese medicine, behavior modification, dietetic and sport adjustment for 22 cases of simple obesity children (7-15 years old, 18 boys, 4 girls) selected for one month, and follow-up survey 6 months after treatment.Results Five thousands,six hundreds and eighty-eight high and primary school students investigated had an overweight incidence of 15.4%, an obesity incidence of 3.2%,boys' overweight incidence and obesity incidence(19.2%,4.6%) were remarkably higher than those of girls (11.5%,1.7% P<0.001). Overweight incidence of various age groups evidently differed(χ2=42.88 P<0.001) with the group of 8-15 years old children as popular. Incidence of various age groups also differed(χ2=21.28 P<0.05) with 7-10 years old and 14-15 years old children as popular. After one-month treatment, weight of all the 22 cases of fat children decreased from (76.45±19.87) kg upon hospitalized to (69.06±17.98) kg with a decrease of (7.43±2.58)kg, BMI value decreased from (31.05±3.96) before treatment to (27.72±3.54).Weight and BMI value before and after treatment differed evidently (t=13.6,12.88 P<0.01), and weight and BMI value decrease were remarkably related with those upon hospitalized (r=0.77 P<0.01;r=0.49 P<0.05).Conclusions Incidence of child obesity has been increasing in recent years, comprehensive treatment consisting of massage intervention along channel of traditional Chinese medicine, behavior modification, dietetic and sport adjustment have good curative effect and are comfortable, well received by children and suitable to promote and apply.

mRNAs of alpha-adrenoceptor (α-AR) subtypes are found in neurons in dorsal root ganglion (DRG) and change after peripheral nerve injury. In this study, the distribution of α-AR subtype proteins was studied in L5 DRG of normal rats and rats with chronic constriction injury of sciatic nerve (CCI). Using immunofluorescence technique, it was found that α1A-, α1B-, and α2A-AR proteins were expressed in large, medium, and small size neurons in normal DRG, and significantly increased in all size neurons 14 days after CCI. α1D- and α2C-AR was also expressed in all size neurons in normal DRG. However, α1D-AR was significantly increased and α2C-AR was decreased in small size neurons 14 days post CCI. α2B-AR neurons were not detectable in normal and CCI DRG. Co-expression of α1A- and α2A-AR in the same neuron was observed in normal DRG and increased post CCI. Collectively, these results indicated that there is distinct distribution of α-AR subtypes in DRG neurons, and the distribution and levels of expression of α-AR subtypes change differently after CCI. The up-regulation of α-AR subtypes in DRG neurons may play an important role in the process of generating and transmitting neuropathic pain.

Tutor system has been widely applied in the cultivation of talents, such as undergraduates, graduate students and young scientist.Scientific research tutor system promotes the interaction between tutors and students, and improves the level of personnel training.In view of the problems of cultivation of the young scientists in hospital, we established a model of cultivation of young scientists by setting mentors.After a preliminary practice, more significant results have been achieved.