ObjectiveTo develop a method of location of prenatal female mouse reproductive tract based on paraffin block serial sections of female fetuses, and get high quality paraffin sections of fetuses containing female reproductive tract (FRT).And to investigate the role of Wnt5a during the development of FRT of prenatal mice. Methods The sex of fetuses at gravidity 15.5 days( G15.5d), G17.5d, G19.5d) and G21.5d was identified by polymerase chain reaction(PCR),and the female fetuses were embedded in paraffin block, the specimen was sectioned serially . One of every four sections was stained by hematoxylin-eosin(HE), and the next one being used later.The images of specimen were taken with digital camera and the serial sections were obtained. The paramesonephric duct /uterus were located and recognized. Immunohistochemistry was used to determine the location and intensity of Wnt5a staining in the middle of the paramesonephric duct / uterus. Results The morphology of the paramesonephric duct /uterus was recognized first and the sections of fetuses with paramesonephric ducts were obtained. The intensity of Wnt5a immunohistochemical staining was increased from G15.5d to G21.5d in mesenchymal cells(P<0.01).Conclusion Wnt5a plays a marked role in early period of female mouse reproductive tract, and is possible to be a key factor to induce uterus differentiation and development.

Objective To study the relationship between serum and cerebrospinal fluid neuron specific enolase (NSE) and myelin basic protein (MBP) concentrations with the disease progress and prognosis in the patients with craniocerebral injury.Methods Forty-five patients with craniocerebral injury treated in our hospital were selected and divided into the mild group,moderate group and severe group according to disease severity;which were divided into the subdural hematoma group,epidural hematoma group,cerebral contusion and laceration group and combined injury group according to the injury types;which were divided into the death and plant survival group,disability group and good recovery group.Other 15 individuals undergoing physical examination were selected as the control group.The concentrations of cerebrospinal fluid and serum NSE and MBP were detected at admission in the patients with craniocerebral injury and control group,on 1,7,14 d after injury in the patients with severe craniocerebral injury.Results The concentrations of cerebrospinal fluid and serum NSE and MBP in the patients with mild,moderate and severe craniocerebral injury were significantly higher than those in the control group,the severe group was significantly higher than the moderate group(P<0.05);the concentrations of cerebrospinal fluid and serum NSE and MBP in the patients with epidural hematoma were lowest,while which in the combine injury group were significantly higher than those in the subdural hematoma group,epidural hematoma group and cerebral contusion and laceration group(P<0.05);which in the death and plant survival group were significantly higher than those in the disability group and good recovery group(P<0.05);which on 1-14 d after injury in the patients with severe craniocerebral injury showed the decreasing trend,but which on 14 d were significantly higher than those in the control group(P<0.05).Conclusion The concentrations of cerebrospinal and fluid serum NSE and MBP are positively correlated with injury severity,which can serve as the basis for diagnosis and prognosis judgment.

ObjectiveTo evaluate the uncertainty for the determination of ethanol in human blood by auto-headspace gas chromatography (HS-GC) with internal standard curve method.Methods Each source of uncertainty, arising from the procedure of testing, was analyzed and conifrmed according to the guidelines of the uncertainty in measurement . After each uncertainty component was quantized, the combined standard uncertainty and the expanded uncertainty of the result were calculated.Results The relative uncertainty brought from the measurement repeatability, standard solution of the ethanol, the sample of blood, internal standard solution of the tert-butyl alcohol, the calibration cure, gas chromatography were 3.4%,0.71%,0.61%,0.41%,1.1% and 1.3% respectively; the relative expanded uncertainty of ethanol in blood was 3.9%.Conclusion The measurement uncertainty of the concentration of ethanol was came primarily from the measurement repeatability of sample, HS-GC and standard curve of ethanol.

Objective To use the colloidal gold immunochromatography (GICA ) method to conduct the methodological prelimi-nary evaluation on pepsinogen Ⅰ (PGI) reagent kit .Methods The method provide by the Preliminary Evaluation of Clinical Quan-titative Experimental Methods :Approval Guide Second Edition (EP10-A2) formulated according to the Clinical Laboratory Stand-ards Institute (CLSI) was used to continuously detect the low ,moderate and high concentrations of serum for 5 d ,Then the related data were collected for analyzing the dispersion degree ,linearity ,offset ,precision and so on .Results The PGⅠ quality control ser-um (concentration 25 ,50 ,100 μg/L) was detected at low ,medium and high concentration levels ,the linear regression equation ob-tained by analysis was Y=0 .9939X+0 .7433 ,correlation coefficient (R2 )=0 .9992 ;the offsets were 0 .37 ,0 .77 ,0 .78 μg/L re-spectively ,total imprecision was 3 .04% ,1 .17% and 1 .08% respectively .Conclusion The GICA related technical indicators of PGⅠreagent kit reach the standards of EP10-A2 document ,the detection results are accurate with high sensitivity and good stability , and conform to the requirements of clinical applications .

Objective: To establish a method for determination of methone and pulegone in Herba Schizonepetae. Methods: The contents of menthone and pulegone were determined by GC (equipped with FID) with HP-5 fused capillary column (5% phenyl methyl siloxane 30m?0.32mm?0.25?m) after the samples were extracted by the solvents. Results: The linear ranges were 0.002-5.0g/L (r=0.9999) for menthone and 0.002-5.0g/L (r=0.9996) for pulegone, the recoveries of menthone and pulygone were 96.30%-103.9% and 95.7%-102.5%, respectively. Conclusion: The method was simple and accurate, which could be applied to the determination of menthone and pulegone in Herba Schizonepetae.

Objective To probe into the strategies for improving the accessibility to orphan drugs for patients of rare diseases in China. Methods Analysis of typical cases of orphan drug use in recent years in China, interviews of hospital administrators and clinical doctors, and analysis of the present health insurance policies in China for orphan drugs, definition of the concept of drug accessibility, clarification of the factors hindering orphan drug accessibility. Results Four factors are found to hinder orphan drug accessibility in such aspects of science and technology, supply, information transfer and medical assurance in China's medicine and healthcare system. These subjective and objective factors affect drug accessibility of the patients of rare diseases, denying them of drug accessibility, of drug use in time, and of affordability of such drugs. Conclusion To raise the orphan drug accessibility in China, it is necessary to define basic concepts and incentive mechanism of rare diseases and orphan drug, design and raise the response mechanism of the medicine and health system in orphan drug supply, build a three dimensional cooperation model between such parties as the government, enterprise and patient, and reduce patients' economic burdern.

Objective To study ovarian development in vitrificatiou of embryos born mice and expression of growth differentiation factor 9 ( GDF-9 ) in its.MethodsThe vitrification recovery embryos (vitrified-embryo group) and fresh embryos (fresh-embryo group) were transplanted into pseudopregnant mice,respectively.The female offspring mice in two groups were sacrificcd on the 3rd,7th,14th,21st,28th and 60th day after birth,the ovarian tissues were taken,6 mice in each time point of each group.The ovarian development was observed by HE staining,the expression of GDF-9 mRNA and protein at each time point of two groups were detected by reverse transcription(RT)-PCR and western blot.ResultsHE staining showed that no abnormal ovarian development was observed in offsprings at two groups.On the 3rd,7th,14th,21st,28th and 60th day after birth,the expression of GDF-9 mRNA in vitrified-embryo group were 0.14 ± 0.07,0.42±0.16,1.00±0.24,1.59±0.28,2.05 ±0.32 and 2.23 ±0.21,respectively,which in fresh-embryo group were 0.13 ±0.06,0.45 ±0.18,1.00 ±0.21,1.56 ±0.26,2.01 0.37 and 2.26 ±0.23,respectively,there was no statistical difference between two groups ( P ＞ 0.05 ) ; the expression of GDF-9 protein in vitrified-embryo group were 0.040 ± 0.030,0.120 ± 0.060,0.170 ± 0.030,0.250 ± 0.040,0.320± 0.060 and 0.330 ± 0.010,respectively,which in fresh-embryo group were 0.030 ± 0.020,0.110 ± 0.040,0.150 ± 0.010,0.210 ± 0.020,0.360 ± 0.070 and 0.350 ± 0.030,respectively,there was no statistical difference between two groups ( P ＞ 0.05 ).Conclusion The ovarian morphology in vitrification of embryos born mice and expression of GDF-9 in ovary has no any obvious change.

Microglia are the inherent macrophages and immune surveillance cells in the central nervous system (CNS) and compose the first guard of immune defense in CNS .The activation of microglia is one of the pathological features of many CNS diseases and acts as an important role during the multiple sclerosis (MS) process.MS is a CNS disease characterized by neuroinflammatory infiltration , demyelination and axonal damage.Accumulation of activated microglia at the injury site has been observed in brains of MS patients and experimental animals with complicated mechanisms.Microglia have both detrimental and beneficial roles .For instance, microglia have been shown to recruit and reactivate T cells in the CNS and release many detrimental molecules such as proteases , inflammatory cytokines, and free radicals.Conversely, they have also been observed to aid in axonal regeneration and remyelination as well as assist in the clearance of inhibitory myelin debris .In addition, microglia have been shown to release a variety of neurotrophic factors . Cuprizone [oxalic acid bis (cyclohexylidene hydrazide )] is a well-known copper-chelating agent.Cuprizone ingestion in mice induces a highly reproducible demyelination of distinct brain regions .Discussion on the detrimental and beneficial aspects of microglia in cuprizone animal models will serve to better understand the development of MS and find out new therapeutic targets.This review will further our understanding of the dichotomous roles of microglia in cuprizone -induced demyelination in animal models of multiple sclerosis .

Objective To evaluate the value and safety of flexible bronchoscopy in etiological diagnosis and treatment of hemoptysis in children. Methods The flexible bronchoscopy findings and clinical data of 22 children with hemoptysis were ana-lyzed retrospectively. Results The causes of hemoptysis were as follows, bronchiolitis and pneumonia in 8 cases (2 cases with bronchiectasis), foreign body in bronchus in 3 cases , idiopathic pulmonary hemosiderosis in 3 cases, neoplasms in airway in 2 cases, suspi-ciion of bronchial pulmonary vessel dysplasia in 4 cases (requiring angiogram) and unknown causes in 2 cases. Two cases suffered hypoxemia. Active bleeding was found in 6 cases and topical hemostasis treatment was effective. The bronchial foreign body was removed from right lower lobe bronchus in 3 cases. One case with atelectasis caused by granulation was treat-ed with bronchoscopic cryotherapy 2 times, no granulation was found during the third bronchoscopy and chest CT showed the atelectasis was cured 2 weeks later. Conclusions Flexible bronchoscopy is one of the necessary and safe examinations in chil-dren with hemoptysis and is of important value in clinical diagnosis and treatment.

Objective:To evaluate p53,c erbB 2,p21 and nm23 oncoprotein expression in diagnosis and treatment of gastric cancer.Methods:The expression of p53,c erbB 2,p21 and nm23 oncoproteins was detected with immunohistochemical method in 63 surgically resected specimens and its endoscopic biopsy specimens of gastric cancer.Results:The positive rates of p53,c erbB 2,p21 and nm23 oncoprotein expression were 37 6%～46 2%,34 6%～56 8%,37 8%～61 5%,70 3%～30 8% respectively.Oncoprotein expression was not observed in non tumor endoscopic biopsy specimens.The expression of c erbB 2,p21 was correlated with grade of tumor differentiation and the expression of p21,nm23 oncoproteins was correlated with the depth of tumor invasion and clinical different stage,namely tumor metastasis.Positive rates of p53,c erbB 2,p21 and nm23 oncoproteins between biopsy and resected specimens was of coincidence.Conclusion:Determination of p53,c erbB 2,p21 and nm23 oncoprotein expression was might be useful in diagnosis and treatment of gastric cancer,differentiating benign and malignant tumor and clinical stages,of gastric cancer. [