BACKGROUND: If the generative cell DNA of each grade is damaged or mutated, it is possible to transmit to the further generations by means of fertilized ova. As a traditional antiepileptic, the mutagenic effects of phenytoin sodium on so matic cells had been confirmed by many researches, but it is still unknown whether phenytoin sodium has the mutagenic effects on generative cells. OBJECTIVE: To detect the mutagenic effects of phenytoin sodium on human sperm chromosomes. DESIGN: In present study we measured human sperm chromosomes in vitro by means of randomized control observa tion. SETTING: Mental Health Center of the First Affiliated Hospital, Chongqing University of Medical Sciences. MATERIALS: Phenytoin sodium was purchased from Sigma Company. The human sperms were collected from the healthy males who had not contacted with any physicochemical mutagen within recent 6 months. Ova were collected from female golden hamsters of 6-8 weeks, which were purchased from Shanghai Institute of Family Planning Science. Medium for washing sperms and ova was the BWW solution containing 0.3% human serum albumin (HSA); Medium for capacitation was the BWW solution containing 3.5% HSA; Medium for post-fertilization was an oval one containing 10% hamster serum. METHODS: After washing, centrifugation and capacitation, the sperms were made into suspension and dispended into 5 centrifuge tubes (5 mL each): Bleomycin A5 (40 mg/L) was added in the first tube as positive control group, phenytoin sodium (10, 20 and 40 mg/L) were added in three tubes respectively, another tube did not contain any reagent as blank control. Hamster ova without pellucid zone were prepared, and equally divided into five portions, which were mixed with the above-mentioned sperms in the five groups respectively, so as to make the hamster ova fertilize, finally human sperm chromosomes were prepared with the fertilized hamster ova. The rate of chromosomal structural aberration (rate of aberrant sperm) and number of chromosomal breakages were examined. We examined the rate of chromosomal structural aberration (rate of aberrant sperm) and number of chromosomal breakages. The effects of phenytoin sodium of three different concentrations on human sperm chromosomes were detected in vitro, and the results were compared with those in the positive control group and blank control groups. MAIN OUTCOME MEASURES: The structural aberration of sperm chromosomes, rate of aberrant sperm and number of chromosomal breakages were observed. RESULTS: ① Chromosomal structural aberration: The structural aberrations of sperm chromosomes including the chro mosomal breakage, monome breakage, fragments, crossing-over aberration, double centromere and ring-like chromo somes were observed in the phenytoin sodium groups, positive control group and blank control group, especially in the phenytoin sodium 40 mg/L group and positive control group. ② Rate of aberrant sperms and number of chromosomal breakages: The rate of aberrant sperms and number of chromosomal breakages were higher in the phenytoin sodium groups and positive control group than in the blank control group, but there was the significant differences between phenytoin sodium 40 mg/L group and positive control group (P ＜ 0.005, P ＜ 0.05-0.01). CONCLUSION: Phenytoin sodium has obvious influence on the structural aberration of human sperm chromosomes and may have mutagenic potential to human sperm cells.

BACKGROUND: Clozapine is a common antipsychotic drug for treating psychotic patients. Some reports suggest that it can cause chromosomal aberration of human cells. This study was designed to analyze the effect on mutagenesis of human generative cells and genetics toxicity of generative cells.OBJECTIVE: To research the effect of clozapine on human sperm chromosome with testing system ex vivo.DESIGN: Randomized controlled observation on the basis of human sperm chromosome.SETTING: Center of Psychological health, the First Hospital affiliated to Chongqing Medical University.MATERIALS: Human sperm was selected from healthy adult males who were not received mutagenesis factors within half a year. Clozapine was provided by the Ninth Pharmaceutical Factory of Shanghai. Ovum was selected from female golden shrewmouse aged 6-8 weeks. Ovum was fertilized and washed with BWW culture medium containing 0.3% or 3.5% human serum albumin. After fertilization, ovum was cultured with ovi-culture medium containing 10% serum of shrewmouse.METHODS: At three days before experiment, shrewmouse was muscularly injected with 40 unit/ampoule preganat mares esrum gonadotrophin, and then with 30 unit/ampoule human chorionic gonadotrophin. Semen was maintained in aseptic beaker and made 5 mL sperm suspension after washing, centrifugation and capacitation. The suspension was equally put into 5centrifuged tubes. 40 mg/L bleomycin A5 was added into one tube to regard as positive control, one tube was regarded as negative control without adding any reagent, and other tubes were added with clozapine at the concentrations of 200, 400 and 800 μg/L, respectively. Ovarium mound cells and pellucid zone in ovum were wiped out with 0. 1% alidase and pancreatin, and then, equally transplanted into a blank gutta in five culture medium. Sperm chromosome was established with stepped fixed air technique.MAIN OUTCOME MEASURES: Sperm rate and broken amount of chromosomal structural aberration.RESULTS: When concentration of clozapine was at 200, 400 and 800 μg/L,respectively, sperm rate and broken amount of chromosomal structural aberration were not significantly different from those in blank control group, and there was also no significant difference among three concentration groups. However, there was significant difference between 40 mg/L bleomycin A5 group and negative control group (P ＜ 0.01).CONCLUSION: Clozapine cannot damage human sperm chromosome through detecting the effect of mutagen on breakage of chromosome with testing system ex vivo, but it has other genetics toxic mechanisms on human sperm chromosome.

BACKGROUND: Heroin, which is characterized by strong liposolubility and immediate effect, can rapidly enter central nervous system through blood brain barrier; however, mechanism of its dependence is still unclear up to now. Therefore, it becomes a hot topic to investigate mechanism of molecular biology of drug dependence, especially to research changes of gene expression.OBJECTIVE: To find out the expression of specific gene related to heroin dependence so as to elucidate the mechanism of molecular biology of heroin dependence.DESIGN: Randomized controlled observation on the basis of experimental animals.SETTING: Center of Psychohygiene, the First Hospital affiliated to Chongqing Medical University.MATERIALS: Eight C57BL/6J mice were randomly divided into experimental group and control group with 4 in each group. Drug and equipments: heroin, Tripure separating agent, PolyATtract mRNA separating system Ⅲ, etc.METHODS: Heroin-dependent models in mice were established by dose-increasing hypodermical injection of heroin for 7 days. On the 7th day,80 mg/kg heroin was injected once, and 2 hours later, 5 mg/kg naloxone was injected'. Mice in control group were injected with the same volume of saline and the same dosage of naloxone. Then, mice were put in glass cage to observe jumping times and numbers of animals which had ptosis, diarrhea, wet-dog movement and trembling of anterior claws within 15 minutes.Mice were immediately sacrificed and total RNA and mRNA were extracted from the brains in experimental and control groups with Trripure separating kit, and then, suppressive subtractive hybridization was proceeded.The hybridization sample was amplified by PCR and analyzed with gel electrophoresis.MAIN OUTCOME MEASURES: Quantity of total RNA and mRNA in brain tissue and different expression of cDNA sections in brain.RESULTS: Mice in each group were involved in the final analysis. ①Results of abstinent symptom after injection of naloxone: Abstinent symptom was observed on mice in experimental group. Jumping times within 15 minutes were 9.75±1.65, which was significant difference from those in control group (P ＜ 0.01). ② Comparison of total RNA and mRNA in brain tissue and different expression of cDNA sections in brain: Quantities of total RNA and mRNA were higher in experimental group than those in control group, and difference of mRNA was significant (P ＜ 0.05). After the second suppressive subtractive hybridization, mixture was amplified with PCR and nest-like PCR to differently express cDNA sections.CONCLUSION: Withdrawal of heroin dependence may cause changes of gene expression, and there is probably an expression of the specific gene in the brain of heroin-dependent mice.

Objective To explore which scoring methods can prevent the ischemic stroke better.Methods 80 padents with atrial fibrillation were randomly divided into two groups,the patients in A group took anticoagulant therapy using CHADS2 score;the patients in B group took anticoagulant therapy using CHADS2-VAS score;follow-up the ischemic stnoke,death and major bleeding event rate.Results 7 cases of ischemic stroke in A group,1 case in B group,the difference was statistically significant ( P ＜ 0.05 ).Conclusion According to CHA2DS2-VAS score for risk stratification,anticoagulant therapy was safe and effective,and did not increase bleeding event rate.

Objective To investigate the correlation between serum levels of folic acid and vitamin B12 and depression. Methods Thirty patients with depression were selected into depression group whose Hamilton de- pression scale score exceeded 17 and/or Hamilton anxiety scale score exceeded 14. Meanwhile, 30 subjects with- out clinically significant depression were selected into control group. Their serum levels of folic acid and vitamin B12 were determined. Zung Self-rating Depression Scale and Self-rating Anxiety Scale were used to evaluate the depression and anxiety. Results The serum levels of folic acid [ (4. 06±1. 61) vs. (6. 48±2. 36) ng/ml, P < 0. 05 ] and vitamin Bl 2 [ (427.96 126. 20) t . (498.46±119.08) pg/ml, P < 0. 05 ] were significantly lower in depression group than those in control group. No correlation was found between Hamilton depression score and serum folic acid level or vitamin B12 leveL Conclusion Patients with depression have lower serum folic acid and vitamin B12 levels than those of the healthy people.

Interleukin(IL),myelocytic-macrophage colony stimulating factor(GM-CSF),interferon(IFN)and other eytokines can promote body's immune response,and inhibit the incidence and development of cutsrleous malignant melanoma,whose experimental research progress provide an important theory for the clinical application.

Congenital heart disease(CHD) is one of the most common birth defects,which is caused by either hereditary or environmental factors,and more commonly by the combination of both.Although CHD is known to be a polygenetic disease,its molecular pathogenesis is still poorly understood.Many genes including the transcription factors have recently been identified to be involved in the development of heart as well as in the pathogenesis of CHD.At the molecular level,this paper reviews the roles of the transcription factors of GATA4/5/6,Nkx2.5,Nkx2.7,TBX1,TBX5,and TBX20,and the interactions among them in the abnormal development of the heart.

Food allergy is the earliest and most common allergic disease in infants that affects children' s growth and development.It is closely related to other allergic diseases with a great increase in prevalence.It is important to learn the mechanisms and related factors,investigate the prevention and treatment,and explore the influence to other allergic diseases with food allergy in infants.

ObjectiveTo observe the influence of mini-invasive evacuation of hematoma for myelin basic protein (MBP) and neuron-specific enolase (NSE) in patients with cerebral hemorrhage.Methods Eighty patients with cerebral hemorrhage were divided into group A and group B by random digits table method with 40 cases each.The patients in group A were given conservation treatment.The patients in group B were given mini-invasive evacuation of hematoma treatment.At the same time 30 cases of healthy people randomly were selected as control group.The serum MBP and NSE were determined before treatment,after treatment 1,3,7,14 d in group A and group B.ResultsThe effective rate in group B was 92.5％ (37/40),which was higher than that in group A [80.0％(32/40)],there was significant difference (P ＜ 0.05 ).The serum NSE before treatment in group A and group B was higher than that in control group [ (12.8 ± 2.9),( 13.1 ± 2.8 ) μg/L vs.( 5.3 ± 2.4) μg/L],there was significant difference(P ＜ 0.05 ).The serum NSE after treatment in group B was obviously decreased compared with group A.The serum MBP after treatment 1 d was increased,7 d was decreased,that in group B was obviously decreased compared with group A [ (5.4 ± 1.5) μ g/L vs. (6.9 ± 1.3) μ g/L,P ＜ 0.05 ].ConclusionThe method of mini-invasive evacuation of hematoma in treatment of cerebral hemorrhage patients has good effect,and can decrease the serum MBP and NSE.

Mutations in CD2AP, mapped to 6pL2. 3 and encoding CD2 - associated protein ( CD2AP), are responsible for autosomal recessive or autosomal dominant steroid - resistant nephrotic syndrome (SRNS). CD2AP plays a key role in the slitdiaphragm network of the kidney, which is necessary for the structure and ultrafiltration functions of the slitdiaphragm. The CD2AP homozygous mutation results in early - onset SRNS while heterozygous expression of the CD2AP mutation has increased susceptibility to glomerular injury.No recurrence of proteinuria was observed in the patient with SRNS with CD2AP homozygousmutationafter the renal transplantation. Therefore, detection of the CD2AP gene in the patients suffering from SRNS will be beneficial to making therapeutic decisions and predicting prognoses.