BACKGROUND: Conventional opinions believe tissues of central nervous system(CNS) cannot regenerate after injury when they have been developed and maturated. However, recent researches have validated that neural stem cells exist in human and adult animal nervous system, and most of them are in static status in vivo. The therapeutic effects of neural stem cells in cerebral infarct have been a key point in the researches.OBJECTIVE: To observe the reactive process of endogenous neural stem cells after cerebral infarct to explore the effects of endogenous neural stem cell in CNS traumatic rehabilitation, which provides theoretical gist for the self-restoration after cerebral infarct.DESIGN: A randomized controlled trial by using experimental animals as subjects.SETTING: Department of Neurosurgery of Peking Union Hospital.MATERIALS: The study was conducted in the Laboratory of Neurosurgery Department of Peking Union Hospital from March to October in 2003. Totally 82 healthy male Wistar rats with a body mass between 250 g and 300 g were selected.METHODS: Cerebral infarct model was established in rats by thread-ligation method. Rats were divided into post-infarct 1 day, 3 days, 7 days, 14 days and 28 days groups with 14 rats each. Control group was sham-operation group( n= 12). The expressions of bromodeoxyuridine (BrdU) and Nestin in the brain of the rats were dynamically detected by immunohistochemical method.MAIN OUTCOME MEASURES: The changes of BrdU and Nestin positive cell numbers after cerebral infarct in rats.RESULTS: Only very few BrdU and Nestin positive cells had survived in hippocampal dentate gyrus and subventricular zone(SVZ) area in control group. BrdU positive cell significantly increased compared with control group in hippocampus and SVZ area after 1 day of cerebral infarct( P ＜ 0.05),which reached its peak on the 7th day( P ＜ 0.05), started to reduce after 14 days but still in the level significantly higher than normal( P ＜ 0.05), and closed to normal level after 28 days. And moreover, BrdU and Nestin positive cells were significantly more in infarct side than the opposite side ( P＜ 0.05), which migrated towards the opposite side through callus.CONCLUSION: Cerebral infarct can activate the proliferation in situ and migration of endogenous neural stem cell.

Objective To investigate whether quantitative dynamic contrast enhanced MR can predict final pathologic response in primary breast cancer patients undergoing neoadjuvant chemotherapy (NAC).Methods Forty seven patients who were pathologically proved infiltrating ductal carcinoma with core needle puncture biopsy were examined before NAC and after 2 cycles of treatment and the quantitative parameters (Ktrans,Kep and Ve) were analyzed prospectively.Histological response is categorized as non-major histological response (NMHR) and major histological response (MHR).Quantitative parameter changes measured after 2 cycles of NAC were compared between MHR and NMHR using non parametric tests (Mann-Whitney U test) and pretreatment parameters were compared using independent samples t tests.Receiver operating characteristic curve (ROC) was used to determine the best predictor and cutoff value.Results Fifteen patients were grouped into MHR and 32 patients were NMHR.Pretreatment parameters(Ktrans,Kep and Ve) were(1.51 ±0.33) /min,(2.97 ± 1.06) /min and (0.55 ±0.16) in MHR and (1.53 ±0.40) /min,(2.82 ± 0.99) /min and (0.57 ± 0.20) in NMHR.There was no significant difference between the two groups (t values were-0.123,0.450 and-0.380,respectively,P ＞ 0.05).Changes inkinetic parameters(K,Kep and Ve) were-88.2％ (-96.0％ to 1.5％),-62.5％ (-94.3％ to -8.7％) and-57.0％(-82.1％ to 55.5％) in MHRand-8.1％(-88.5％ to 32.4％),-18.2％ (-62.1％ to 145.9％) and-4.7％ (-83.1％ to 95.7％) in NMHR.There were significant difference between the two groups (Z values were 4.359,4.359 and 3.332,respectively,P ＜ 0.01).The areas under ROC curve of AKtrans,AKep and Ktrans after 2 cycles of NAC were all 0.898.Sensitivity of the three parameters for predicting NMHR were 87.5％,90.6％ and 78.1％,and specificity were 86.7％,80.0％ and 93.3％,respectively.Conclusion Quantitative dynamic contrast enhanced MRI can predict final pathologic response in primary breast cancers after 2 cycles of NAC.

Objective To establish a method for detection folylpolyglutamate syntbetase (FPGS),explore the change of FPGS in the drug-resistant A549 cells induced by methotrexate(MTX) enantiomer,and provide new tools to further investigate drug resistant mechanism. Methods A549 cell lines induced by L-( + )-MTX and D-( - )-MTX (25 μmol/L) were chosen to raise three cell lines as compared with MTX-sensitive cell line. Then FPGS were extracted for the CEIA-LIF and western blot was performed. After validation, FPGS antibodies were labeled by fluoreacein isothiocyanate (FITC) and produced a immune response with former-extracted FPGS. CELA-LIF can separate and detect labeled proteins according ruination time of the protein with different size and detect FPGS in drug resistant cell lines induced with L-(+)-MTX and D-(- )-MTX. The accuracy was evaluated as compared with western blot assay. Results The separation time of CEIA-LIF for labeled FPGS antibody and the immune complexes were7. 1 min and 8.9 min, and the resolving power was 4. 5. The process of protein separation and detection can be accomplished in less than 10 minutes. Western blot analysis showed there was no non-specific bands appears in the extract of these three cell lines after the freeze-thaw in liquid nitrogen. The minimum detection level in sensitive cell strains was 0. 68 mg/μl. The consent of FPGS in L-(+)-MTX and D-( - )-MTX induced cells were 46. 59% and 48. 36% compared with drug sensitive cell strains with CELA-LIF. Conclusions CELA-LIF was established in this experiment. It is efficient and sensitive for detecting of FPGS, which is similar to western blot method. The level of FPGS in L-( + )-MTX and D-( - )-MTX induced drug resistant cell lines is significantly lower, indicating the expression of FPGS is damaged.

Objective Analyzing the construction process and application of health appropriate technology database in Zhejiang province and proposing related considerations and recommendations for the database construction.Methods To design and construct the technology database system,and using excel software to calculate statistic data.Results Established a health appropriate technology database,which contains 826 technologies that can be selected and promoted by the demonstration bases in the whole province.Conclusions The establishment of the technology database increased the efficiency of health appropriate technology transformation at the provincial level,and also provided a reference for the national-wide generalizing of health appropriate technology.

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Objective To investigate the value of X-ray guided desmopressin (DDAVP) stimulated bilateral inferior petrosal sinus sampling (IPSS) in diagnosing the recurrence of ACTH dependent Cushing disease or ineffectiveness after surgery or radiotherapy. Methods Retrospective analyses of patients with recurrent ACTH dependent Cushing disease (31 cases) or ineffective (3 cases) treatment after surgery or radiotherapy from January 2013 to October 2014 in our hospital was conducted. Bilateral inferior petrosal sinus angiography showed the same side of the cavernous sinus to prove successful intubation. The cases with discontinuous of the inferior petrosal sinus and cavernous sinus were excluded by this study. Finally, there were 34 cases of the patients in this study. Diagnosis was based on the ratio of ACTH level in IPS to peripheral vein after desmopressin test.The gradient≥2 at baseline or gradient≥3 after desmopressin test suggested the sources were in the pituitary. Diagnosis was confirmed by gold standard to investigate the value of X-ray guided desmopressin (DDAVP) stimulated bilateral IPSS. Results The IPS gradient≥2 at baseline or gradient≥3 after desmopressin test suggested the sources were in the pituitary in 30 patients. A total of 22 (22/30) patients underwent surgery with a final diagnosis of ACTH adenoma. The symptoms were obviously relieved in 8 (8/30) cases after sellar area gamma knife treatment and lesions were confirmed in the pituitary. IPS gradient<2 at baseline or gradient<3 after desmopressin test was found in 4 cases. One case (1/4) was found to have for ACTH adenoma after pituitary surgery. The other 3 cases (3/4) were confirmed to have lung carcinoid and clinical symptom alleviated after surgery. The sensitivity of desmopressin stimulated IPSS was 96.8%, the specificity was 100%, and the accuracy was 97.1%. Conclusion Desmopressin stimulated IPSS is an effective diagnostic procedure in diagnosing ACTH dependent Cushing disease recurrence or ineffectiveness after surgery or radiotherapy.

Objective To grasp the general situation of patent application and authorization of 77 medical and health institutions in Zhejaing,to provide reference and basis for promoting the patent application and conversion.Methods Patent data were searched through Bai Teng net patent database,and SPSS 19.0 was used for statistical analysis.Results The total number of patent application showed a trend of generally rise.There were 1430 valid invention patents in total,among which 264 were invention patents and 1166 utility model patents.The total number of patens as well as invention patents were positive correlated to province GDP GDP and doctors per thousand population (r=0.824,0.812,0.784,0.771 respectively,P＜0.001).The rate of collaborative patent application was 6.28％,and there was a significant difference (α=0.05/3,P＜0.001) in the collaborative patent applications around Hangzhou,provincial and other cities medical and health institutions.Conclusions The total number of patents in 77 medical and health institutions in Zhejaing has reached a certain scale,and the number of patents increase with the economic development and the number of doctors per thousand population.The effective patents present in concentrating in regions,and the proportion of invention patents was low.

AIM:To determine the biological feature of circulating endothelial cells (CECs) in acute promye-locytic leukemia ( APL) patients before and after treatment , and to analyze the relationship between CECs and the clinical characteristics .METHODS: The CECs were sorted from peripheral blood by magnetic-activated cell sorting and then counted by 3-color flow cytometry.The cells were identified by immunofluorescence staining for the expression of CD 146, CD31, CD144, VEGFR-2, CD45 and CD133.The CECs were cultured in vitro, and the tube formation and colony-forming rate were determined .RESULTS:Increased quantity of CECs was observed in CD 34 positive group and group with WBC >10 ×109/L (P<0.05).The quantity of CECs had a significant difference among low risk , medium risk and high risk groups (P<0.05).The positive rate of CD133 and quantity of CECs significantly reduced in 32 APL patients when they gain complete remission after treatment (P<0.05).The amount of tube formation and colony-forming rate were significant-ly reduced after treatment (P<0.05).The ratio of CECs quantity from APL patients after treatment to that before treatment had a negative correlation with arsenic concentration in urine on day 7 during As2O3 treatment (P<0.05).CONCLU-SION:Accurately counting CECs may be helpful for evaluating prognosis and designing treatment strategy .

Objective:Cushing′s disease(CD) is caused by the pituitary adrenocorticotroph hormone(ACTH) secreting adenomas, leading to increased serum cortisol levels and various abnormal metabolic processes. Untreated CD is linked to high mortality, thus it is critical to elucidate its pathogenesis. This study aims to explore the pathogenesis of pituitary ACTH adenomas using whole-genome sequencing analysis.Methods:Fresh tumor tissues and peripheral blood samples were collected in 9 confirmed cases of pituitary ACTH adenomas who underwent surgery. Whole genome sequencing was then performed, followed by analysis and verification of single nucleotide mutations, copy number variation(CNV) and chromosome structure variations.Results:Somatic USP8 mutations(p.Ser718del, p. Ser718Pro, p. Pro720Arg, p. Pro720Gln) were found in 5 patients, with a rate of 55.6%; CNV of USP8 was detected in 1 patient; TP53(p.Cys135Tyr), NF1(p.Val1049Glufs*11) and KMT2C(c.3323+ 1G>A) mutations were identified in 1 patient harboring wild-type USP8. CNV analysis showed a loss of heterozygosity in multiple chromosomes in a wild-type USP8 patient. Structural variations were found in 2 with unknown significance. No germline gene mutations were detected in this study.Conclusion:Somatic USP8 mutations, increased copy number of USP8, variations of tumor-related genes such as TP53 and extensive somatic CNV all contribute to pathogenesis of CD. Chromosomal structure variations may suggest high-risk pituitary ACTH adenomas, and call for frequent follow-up and aggressive treatment.