Objective To evaluate analytical performance of homocysteine (Hcy) by circulating enzymatic method .Methods Re‐ferring to CLSI evaluation project and pertinent literature ,and by combining our actual works .we designed a verification procedure and experimental method .By using these above ,the precision ,accuracy ,analytical measurement range ,clinical reportable range of Hcy by circulating enzymatic method were evaluated .Results would be compared with the declaration of the manufacturer (NingBo Medical System Biotechnology Co .,Ltd) or desirable specifications derived from biologic variation .Results The results showed that the within‐run CV were 1 .26% and 0 .84% ,and the total CV were 1 .36% and 1 .32% ,less than 10% of the manufacturer′s statement .The relative bias between the results measured for calibrator at tow levels and target value was 3 .69% and 0 .69% ,less 10% .AMR was 3 .38-51 .81 μmol/L ,and the most suitable dilution rate was 1∶3 ,so the CRR was 3 .38-155 .43 μmol/L .Con‐clusion The analytical performance of Hcy analyzed by circulating enzymatic method is consistent with the standards which manu‐facturers has proclaimed ,so it is conform to the requirements of clinical .

Objective Several studies have indicated that miR-15a,miR-15b and miR-16 may be the important regulators of apoptosis.Since attenuate apoptosis could protect myocardium and reduce infarction size,the present study was aimed to find out whether these miRNAs participate in regulating myocardial ischemia reperfusion (I/R) injury.Methods Apoptosis in mice hearts subjected to I/R was detected by TUNEL assay in vivo,while flow cytometry analysis followed by Annexin V/PI double stain in vitro was used to detect apoptosis in cultured cardiomyocytes which were subjected to hypoxia/reoxygenation (H/R).Taqman real-time quantitative PCR was used to confirm whether miR-15a/15b/16 were involved in the regulation of cardiac I/R and H/R.Results Compared to those of the controls,I/R or H/R induced apoptosis of cardiomyocytes was significantly iucreased both in vivo (24.4％ ± 9.4％ vs.2.2％ ± 1.9％,P ＜ 0.01,n =5) and in vitro (14.12％ ±0.92％ vs.2.22％ ± 0.08％).The expression of miR-15a and miR-15b,but not miR-16,was increased in the mice I/R model,and the results were consistent in the H/R model.Conclusions Our data indicate miR-15 and miR-15b are up-regulated in response to cardiac I/R injury,therefore,down-regulation of miR- 15a/b may be a promising strategy to reduce myocardial apoptosis induced by cardiac I/R injury.

With the widespread use of multi-slice spiral computed tomography(CT) and the increasing level of diagnosis and treatment,the incidence of pulmonary ground glass opacity(GGO) is increasing.The etiology of GGO is complex and the pathological types are diverse.In addition to benign lesions,GGO can be a specific type of lung adenocarcinoma or its invasive lesions.Some scholars think it is closely related to early lung adenocarcinoma.Histopathology of lung cancer is crucial for the prognosis and treatment of lung cancer.However,under normal circumstances,the pathology of the patient can be known by surgery,bronchoscopy or puncture.However,many patients may not have the pathological conditions for obtaining the pathological specimen.CT as the primary method of diagnosis of lung adenocarcinoma,lung adenocarcinoma in the new classification,based on the lung GGO CT findings to determine its ability to provide a characteristic indicator of tumor histopathology in order to help GGO diagnosis and guidance of clinical treatment.

Objective:In this study, we established a reliable surgical procedure of lung ischaemia-reperfusion(IR) injury in rats. The research progress of different lung IR injury models and application value was also discussed.Methods:Twenty-eight adult SD rats were randomly divided into SHAM group and lung IR injury group(IR group), 14 rats in each group. In IR group, rats underwent tracheotomy under general anesthesia and received mechanical ventilation. Chest was opened in supine position, and pulmonary hilum was blocked for 30 minutes then the occlusion was removed. Samples were harvested after reperfusion for 45minutes. Rats in SHAM group received surgery and exposure of the right pulmonary artery, and experienced the same amount of time before the chest closed. Arterial blood gas was extracted postoperatively. Gross view of the lungs and pathological changes were observed, and the dry/wet ratio(W/D) was determined. Protein level of pro-inflammatory factors, markers in oxidative stress pathway, and endothelial functional markers in lung were tested by western blot analysis.Results:In IR group, there was pink foamy secretion in the airway, and the lungs exhibited signs of edema and congestion. In IR group, the alveolitis score was significantly increased, the W/D ratio was also increased, p38MAPK and NF-κB signaling pathways were activated, and the expression of TNF-α was significantly increased, while the expression of eNOS was significantly decreased.Conclusion:Left hilum clamping and bilateral reperfusion injury in lung is a practical animal model, it is a simple, low-cost and repeatable animal model for further studies. No microsurgical instruments were required during the procedure. Lung IR injury is characterized by oxidative stress response, inflammatory response and endothelial cell dysfunction.

To observe the incidence of varicella vaccine after breaking through the case of varicella vaccine , immunization strategy ,popularized in the city.Inoculation Population living in the Binhai New Area of full age to 12 years old children in December,has been vaccinated or who have had chickenpox varicella vaccine except .Controls were four districts around the city girls without varicella in children.Methods: Implementation of vaccination for the target population.All vaccinees was observed from 42 days to 2 and a half years later ,the incidence of varicella break cases.In the observation group and the control group was observed in two groups of varicella vaccine protection rate calculation.Results: The gelatin free attenuated varicella vaccine breakthrough in 134 cases,the incidence rate was 0.35%;no gelatin attenuated varicella vaccine protection rate of 80.92%, with domestic and foreign reports consistent.Conclusion:After vaccination from 42 to 2 and a half years ,varicella vaccine can effectively protect children from the onset, while reducing the prevalence of children 's pain and the economic burden of the family.Varicella vaccine is still a breakthrough occurred ,therefore to consider two inoculations ,with further observation of two times after inoculation the body to produce antibody level and epidemiological protection effect.

Objective To compare a novel mirror-overlap method with the traditional manual measurement and the subjective assessment in assessing breast symmetry after oncoplastic surgery.Methods 59 patients with breast cancer who underwent conservation/reconstruction surgery were recruited.The post-operative assessment for breast symmetry was conducted using manual measurement,subjective assessment by the patient,their family member and a nurse,and the mirror-overlap method respectively.The latter method involved using the Photoshop to assess the area differences in breast outlines and shadows created by a flashlight when bilateral images were overlapped in the axis of midline.Results The distance differences between bilateral breasts measured using manual method were negatively correlated with symmetric coefficients in the mirror-overlap method (P＜0.05).There was a positive correlation between the average scores in the subjective assessment and symmetric coefficients of mirror-overlap method (P＜0.05).Conclusion The mirror-overlap method is an easy,feasible and cheap method which collects 3D breast information and provides an effective approach to assess breast symmetry after oncoplastic surgery.

Objective To investigate a more pedect method for a nice outward appearance of a reconstructed thumb.Methods A free one-stage plasty second toe transfer for thumb reconstruction by interchanging the whole skin-nail flap from the great toe with another one from the second toe.Results There were 12 cases in this group,following-up 6-9 months in 8 cases,7 cases was excellent and 1 cases was good.The reconstructed thumb got a nice looking and more normal function while no blight to the great toe occurred.Conclusion It is an effective new procedure in ameliorating outward appearance of the reconstructed thumb by transferring the free moulded second toe.

OBJECTIVE: To further investigate the mechanisms of hair cell generation or regeneration, the primary culture systems of cochlear sensory epithelial cell (CSEC) of rats were established. METHOD: Cochlear sensory epithelial (containing the organ of Corti) of postnatal day 1 (P1) rats was isolated by mechanical dissociation. The explants were grown on sterilized disposable plastic dishes,cultured in Dulbecco modified Eagle medium(DMEM), and observed daily by inverted microscopy. The culture medium was changed twice a week. The pure CSEC was harvested by the limiting dilution method. CSEC was identified by immunocytochemical method with cytokeratin 18, vimentin, Brn3. a, Calretinin, BrdU and ultrastructural examination with transmission electron microscopy. The markers of epithelial cell and the markers of hair cell were used to identify the origin and character of CSEC. CSEC mitotic division was detected by BrdU staining of nuclear DNA. RESULT: The fresh explants were light yellow. The morphology of CSEC couldn't be seen clearly under the inverted microscope because of the complex structures of cochlear sensory epithelial. CSEC grew out of the explants usually at 2nd culture day. Fibroblast like cells (FLC) around the CSEC grew faster than CSEC, but could easily be excluded. Pure CSEC may grow into monolayer with 'cobblestone-like' appearance and show a large, flat, polygonal epithelial morphotype with big, round nuclei. Some cells showed 'Dome' formation, probably due to fluid collection underneath the cell monolayer. The culture CSEC coexpressed cytokeratin 18 and vimentin has rich microvilli and complex tight junction,which indicated the epithelial origination of CSEC. Coexpressed of the Brn3. a and Calretinin of the hair cell characteristic markers suggested the culture cell may represent rat progenitor hair cell. BrdU staining showed CSEC produced new hair cell-like cell (progenitor hair cell) by the mitotic division. CONCLUSION The primary culture systems of cochlear sensory epithelial cell of rats were successfully established in vitro. CSEC coexpressed the characteristic markers of the immature hair cell,which identified the culture cell may represent progenitor cell of rats hair cell. It may be a suitable model for in-depth investigation the molecular mechanisms of hair cell generation or regeneration.
Animals ; Cell Culture Techniques ; methods ; Cell Division ; Cells, Cultured ; Cochlea ; cytology ; Epithelial Cells ; cytology ; Hair Cells, Auditory ; cytology ; Rats ; Rats, Wistar

Objective:To analyze the law of distant metastasis in patients with small lung adenocarcinoma with different CT findings, and to explore the feasibility of different preoperative examination methods for small lung adenocarcinoma with different imaging characteristics.Methods:Clinicopathological data of cT1a-cN0 lung adenocarcinoma patients admitted to the respiratory center of China-Japan Hospital from January 2017 to December 2018 were retrospectively collected. A total number of 785 patients were included, including 289 males and 496 females. SPSS 22.0 was used for statistical analysis.Results:A total number of 785 patients were included in this study, including 287 pure ground-glass nodule (GGN) patients, 111 GGN predominant patients, 221 solid predominant patients and 166 solid nodule patients. Among the included patients, 8 had distant metastasis, including 6 with bone metastasis, 1 with brain metastasis and 1 with brain and adrenal metastasis. No distant metastasis was observed in the patients with pure GGN and GGN predominant nodule, while 1 solid predominant patients had distant metastasis, and 7 patients with solid nodules had distant metastasis. The probability of distant metastasis was 0.5% for the solid predominant patients and 4.2% for the solid nodule patients. Univariate analysis results showed that CEA level ( P=0.030), the largest diameter of the lung window tumor ( P=0.003), the largest diameter of the solid component of the lung window tumor ( P<0.001), the largest area of the lung window tumor ( P=0.002), mediastinal window tumor maximum area ( P<0.001), CTR ( P<0.001), TDR ( P<0.001), and pleural indentation sign ( P=0.037) were risk factors for distant metastases. Multivariate analysis showed that CEA ( OR=1.019, 95% CI: 1.002-1.037, P=0.028) and TDR ( OR=0.000, 95% CI: 0.000-0.310, P=0.001) were independent risk factors of distant metastasis. Conclusion:For patients with pure GGN and GGN predominant nodule, preoperative examination could not be required, but for pure solid nodules, it is necessary to perform relevant preoperative examination including skull MRI before surgery to exclude distant metastasis.

Objective To evaluate the value of endobronchial ultrasound-guided transbronchial needle aspiration in the early diagnosis of non-central small cell lung cancer.Methods 141 patients wereselected from July 1999 to June 2017,who were diagnosed with small cell lung cancer (stage N2 and N3).They were divided into two groups according to the approach of obtaining histopathological tissuefor diagnosis.49 cases in the experimental group were obtained by EBUS-TBNA,92 cases in the control group were done by video assisted thoracic surgery (40 cases,43.5％),mediastinoscopy (1 case,1.1％),and open procedure (51 case,55.4％).Survival outcome,time of waiting for the treatment and lengthof stay were analyzed.Results There werel09 males,32 females,the range of age is from 16 to 79 years old [(56.21 ± 11.48) years].62 lesions located in the upper lobe,12 in the middle lobe,56 in the lower lobe,11 in the middle-lower/-upper lobes.Considering the T stage,there are 7 cases of T1,61 cases of T2,45 cases of T3,and 26cases of T4,2 patients with Tx stage.Compared with conventional approach,EBUS-TBNA saved 10.78 days from admission to the time of receiving treatment [(4.62 ± 0.66)days vs.(15.40 ± 1.61) days,P ＜ 0.05],and saved 18.13 days of length of stay [(5.30 ± 0.76) days vs.(23.43 ± 2.44) days,P ＜0.05].5-year survival rate was 31.0％ for traditional group and 27.5％ for EBUS-TBNA group,there was no significant difference between two groups(P =0.308).Conclusion EBUS-TBNA couldsave the waiting days from admission to the time of receiving treatment,and also shorten the total length of stay.EBUS-TBNA was more efficient than conventional approaches (VATS,mediastinoscopy or open procedure) for diagnosing non-central small cell lung cancer with enlarged mediastinal lymph nodes.