Vitreoscilla Hemoglobin(VHb), with the function of increasing the growth of and product yield by a heterologous host, has been widely use in the area of fermentation, environment protection, transgenic animal and plant, recombinant protein expression, etc. Fusion protein of VHb with other enzyme or protein can enhance activity and stability of the enzyme or isolation efficiency of the protein. The reconstitution of VHb will be helpful to obtain ‘novel’ proteins which have better activity.

BACKGROUND:Electromagnetic fields can cause changes of the body,especially the nervous system.Effect of pulsed electromagnetic fields(PEMFs)on neural stem cells has been detected.OBJECTIVE:To investigate the effect of prenatal pulsed electromagnetic fields(PPEMFs)on neural stem cell proliferation and nestin protein expression in the hippocampus of rat offspdng.METHODS:Sprague Dawley female rats weighing 240-260 g were included and randomly divided into two groups:control and PPEMFs.Rats from the control group were given no interventions.Rats from the PPEMFs group were given PEMFs stress at gestational days 14-20.Each stress was given three times daily for 10 minutes.The male and female offspring rats were sacrificed at 1 month of age and their brains were sectioned to determine the expression of nestin protein and Brdu-positive cells in the hippocampus by immunohistochemistry.RESULTS:The expression of nestin-and Brdu-positive cells in the hippocampus of female and male PEMFS offspring were significantly higher compared with the control group(P<0.001),and there was a significant difference between female and male offspring(P<0.001).The nestin-and Brdu-positive cells in female offspring outnumbered those in male offspring(P<0.001);however,there was no significant difference between female and male offspring in the control group(P>0.05).CONCLUSION:PPEMFs can increase the number and proliferative capability of the neural stem cells in offspring.It may be a pdmary stage of the cascade reaction of the body to the brain damage caused by PPEMFs stress.

Objective To observe the effect of different thyroid hormone level on the expression of synaptotagmin Ⅰ(Syt Ⅰ) in adult rat hippocampus. Methods All 28 adult male SD rats were assigned randomly into hypothyroid, hyperthyroid and control group, hypothyroid group was established by daily intraperitoneal injections with propylthiou raci(PTU, 10.0 mg/kg body weight) for 6 weeks and hyperthyroid group with L-Thyroxine (L-T4, 0.5 mg/kg body weight) for 3 weeks. Radioimmunity method was used to assay the levels of serum T3 and T4, immunohistochemical S-P technology to assay the levels of Syt Ⅰ protein in hippoeampus CA1, CA3 and dentate gyrus (DG). The layers analyzed in the different subfields include the polymorphic cell layer(the stratum oriens, SO), pyramidal cell layer(PCL), stratum radiatum (SR), lacunosum-molecular layer (SLM) in CA1 and CA3, granular cell layer(GL) and molecular layer(ML) in DG. Results The levels of serum T3 and T4[(0.34±0.12), (41.03± 11.37)nmol/L]in the hypothyroid rats were significantly lower than those in the control group[(0.65±0.15), (55.20±10.68)nmol/L, P < 0.01 or < 0.05], and the positive granule of Syt Ⅰ was significantly lower in PCL and SR of CA1 and CA3, GL of DG. The average optical value responsible for Syt Ⅰ immunoreactivity was obviously reduced in SO(0.048±0.007), PCL(0.299±0.035), SR(0.042±0.007), SLM(0.038±0.006) of CA1, PCL(0.085± 0.019), SR(0.040±0.011), SLM (0.038±0.006) of CA3, GL (0.076±0.019) of DG than normal controls (0.068± 0.014, 0.376±0.053, 0.053±0.008,0.056±0.009,0.118±0.026,0.052±0.010,0.053±0.009,0.099±0.015; P< 0.01 or < 0.05). Serum T3 and T4 levels [(1.43±0.30), (157.18±19.95)nmol/L]of hyperthyroid rats were significantly higher than those of control group(P < 0.01). The value was reduced in PCL(0.322±0.050), SR(0.039±0.006), SLM (0.042±0.006) of CA1, PCL(0.098±0.034), SR(0.046±0.013), SLM(0.046±0.010) of CA3 and GL(0.085± 0.024), ML (0.042±0.009) of DG (P < 0.05 or < 0.01). Conclusion Adult-onset of hypothyroidism and hyperthyroidism can reversibly decrease the expression of Syt Ⅰ in CA1, CA3 and DG regions of hippocampus.

To improve the growth enhancement activity of Vitreoscilla hemoglobin(VHb), Vitreoscilla hemoglobin gene(vgb) was mutated by error-prone PCR and then reconstituted by DNA shuffling. The shuffling library was constructed by inserting the shuffled genes into the downstream of vgb natural promoter and transforming them into E.coli DH5?. Mutated active VHb proteins were first screened in test tubes according to host cell pellets color and then in shake flasks according to host pellets wet weight .One active mutant protein, VHb′042506, was obtained after second screening. It could increased the host wet weight by 31.25% and 58.75% than that of the control which bearing natural VHb under microaerobic and extremely microaerobic conditions, respectively. Sequencing and alignment results showed that 11 nucleotides were mutated, thus resulted in 4 amino acids changes occurred in this mutant protein. CO difference spectrum test also indicated that it had higher specific absorption.

AIMTo prepare dilitazem hydrochloride delayed-onset, sustained release tablet, which can not only provide the delay in release start, but also the constant release rate after a lag time. To analyze release mechanism and investigate the effect of outershell compositions on release behavior.

METHODSThe delayed-onset, sustained release tablets were prepared by dry-compression coated technology. The release profiles of uncoated cores and presscoated tablets were compared. Two parameters, time-lag (Tlag) and release rate (k), were used to evaluate the influence of factors, such as the amount of hydroxypropylmethylcellulose (HPMC) and poly-vinyl-pyrrolidinone (PVP) K30, the viscosity of ethylcellulose (EC) and HPMC, and the compression load on diltazam hydrochloride (DIL) release. Higuchi equation and Peppa's equation were used to analyze release mechanism.

RESULTSWith the increase of HPMC amount or HPMC viscosity, Tlag was prolonged and k was decreased; With the increase of PVP K30 amount, Tlag was shortened and k was increased; EC viscosity and compression load above certain degree showed no effect on DIL release.

CONCLUSIONThe drug release from delayed-release tablet is controlled by erosion mechanism, Tlag is determined by the erosion rate of outer-shell.

Cellulose ; chemistry ; Delayed-Action Preparations ; Diltiazem ; administration & dosage ; Lactose ; analogs & derivatives ; chemistry ; Methylcellulose ; analogs & derivatives ; chemistry ; Oxazines ; Polyvinyls ; chemistry ; Pyrrolidinones ; chemistry ; Tablets ; administration & dosage ; chemistry ; Technology, Pharmaceutical ; Viscosity

Objective To apply continuous subcutaneous pulse infusion of gonadotropin-releasing hormone (GnRH) to treat patients with central secondary amenorrhea,and to observe the induction of spontaneous ovulation and natural fertilization.Methods Using micro-pulse infusion pumps with each 90-minute infusion of LHRH 10 μg daily,the serum LH,FSH,estrogen,and progesterone levels ; and change in endometrium and ovarian size before and after treatment were monitored.The end point is natural pregnancy.Results The levels of serum LH,FSH,and estrogen were obviously increased after 4 weeks of treatment.The natural menstrual cycle was rebuilt after 8 weeks of treatment.The patient was pregnant and the treatment was stopped by 16 weeks.The experience of pregnancy was favourable.40 weeks later,the patient delivered a healthy female infant via caesarean section.Conclusions It was the first time in China that the technology of micro GnRH pump was applied in patients with secondary central amenorrhea.The result demonstrates that this technology can perfectly simulate the physiology of hypothalamic GnRH secretion.At present,more patients are included in this research to confirm the effectiveness.

BACKGROUND: Hippocampus is an important region involved in learning and memory. Some methods for freshly isolated hippocampal pyramidal neurons have been described. These methods need multiple enzymes or the procedure is complex.OBJECTIVE: To acquire large quantities of hippocampal neurons from rats suitable for patch clamp study.DESIGN: An animal experimental observation.SETTING: Department of Physiology and Pathophysiology, School of Medicine, Xi'an Jiao Tong University.MATERIALS: The experiments were carried out in the Medical Experimental Center, School of Medicine, Xi'an Jiao Tong University between March and October 2004. The animals were the 10-15-day-old SpragueDawley rats, bothmale and female.METHODS: Hippocampal pyramidal neurons were freshly isolated. Delayed rectifier potassium current and voltage-gated Ca2 + currents were recorded by whole-cell patch clamp configuration.MAIN OUTCOME MEASURES: Acute isolated neurons were observed with a Leica microscope. Recordings of delayed rectifier potassium currents and voltage-gated Ca2+ currents were obtained.RESULTS: ① Under the inverted microscope, the acute isolated neurons had a smooth and glassy surface, a three-dimensional contour, a pyramidal shape with a longer apical dendrite and several basal dendrites. ② After the whole cell recording configuration was formed, voltage-gated barium currents through calcium channels were elicited by 200 ms depolarizing potential from -70 mV to +20 mV, with an increment of 10 mV, a holding potential was -90 mV. Delayed rectifier potassium current (Ikd) was elicited by a protocol consisting of a multiple depolarizing pulses (-60 mV to +50 mV),preceded by a single prepulse step to -50 rmV, with inactivated transient outward potassium currents from a holding potential of -80 mV.CONCLUSION: This method is simple and ideal for the dissociation of neurons from rat hippocampus, and offers a powerful tool for functional analysis of ion channels by patch clamp.

AIM To optimize the decolorization technique for Boletus edulis polysaccharides and to evaluate the antioxidant activity.METHODS With decolorization temperature,resin consumption,polysaccharide concentration and decolorization time as influencing factors,decolorization rate and polysaccharide recovery rate as evaluation indices,orthogonal test was applied to optimizing the decolorization technique on the basis of single factor test.Then the antioxidant activity was determined by Fe2+ chelating power,hydroxyl free radical scavenging activity and reducing power tests.RESULTS The optimal conditions were determined to be 30 ℃ for decolorization temperature,7.5 g/100 mg polysaccharides for S8 resin consumption,1 g/L for polysaccharide concentration,and 2 h for decolorization time,the decolorization rate and polysaccharide recovery rate were 89.34％ and 49.26％,respectively.The polysaccharides demonstrated strong Fe2+ chelating power and hydroxyl free radical scavenging activity with weak Fe3+ reduction power.CONCLUSION This stable and reliable method can be used for the decolorization for Boletus edulis polysaccharides with good antioxidant activity.

Objective To investigate effect of Wenjingtongluo prescription combined with acupuncture and moxibustion on ESR, Fib and hemorheology in patients with cervical spondylosis.Methods 110 cases of cervical spondylosis were divided into two groups, 55 cases in each group.The control group was treated with acupuncture and moxibustion.Experimental group on the basis of acupuncture treatment, were given Wenjingtongluo prescription.The PRI index, VAS score and blood rheology of the two groups were compared.Results The total effective rate of the experimental group was significantly higher than that of the control group (92.73% vs 76.36%) .There was a significant difference (χ2 =5.636, P <0.05) .After treatment, the two groups of PRI index ( emotional score, sensory score, total score ) , VAS score were significantly reduced ( P <0.05 ) .After treatment, the PRI index ( sensory score, total score) and VAS score of the experimental group were significantly lower than those of the control group after treatment.The difference was statistically significant(P<0.05).After treatment, two groups of ESR, Fib, PCV, whole blood viscosity, whole blood viscosity decreased significantly( P<0.05).The experimental group after treatment, ESR, Fib, PCV, whole blood viscosity, whole blood viscosity was significantly lower than the control group after treatment.The difference was statistically significant(P<0.05).Conclusion Wenjingtongluo prescription combined with acupuncture can significantly improve the clinical symptoms, reduce the pain of patients and improve the level of blood rheology.

OBJECTIVE To explore the relationship between urinary tract infection(UTI) and urological surgical site infections(SSI).METHODS The surgical site infections of 115 patients underwent urological operations were observed.Patients were screened for the presence of UTI before the operation and subcutaneous swabs for culture were collected at the end of the operation by brushing with a sterile cotton-swab just before skin closure.The resistance and similarity of the isolates were analyzed by antimicrobial susceptibility test and pulsed field gel electrophoresis(PFGE).RESULTS Surgical site infection occurred in 14 of 115 patients.The isolates from infected surgical sites showed higher resistance to antimicrobial agents than those from non-infected surgical sites(P