Objectives: To observe ultrastructural changes of the corneal endothelium and to assess the significance of the changes after irrigation and aspiration(I/A) of the anterior chamber in rabbits.　Methods: Corneal thickness and intraocular tension were measured and the ultrastructure of corneal endothelium was observed in scheduling times following I/A.　Results:Obvious injury of the membrane structure of corneal endothelium was viewed with scanning electron microscope 1 day after injury. Transmission electron microscope revealed marked swelling of the organelles, increasing vacuoles in the cytoplasm and swelling, denuding, dissolving of the cellular nuclei 3 days after injury. The lipoballs resulted from aggregation of lipid in the cellular membrane adhered to the defect region of endothelium 5 days after injury. Scanning electron microscope displayed marked sign of migration of the corneal endothelium 7 days following injury. The corneal endothelium was heteromorphic and the degeneration of the epithelium occurred. The corneal thickness gradually increased and directly related to the degree of the corneal muddiness. The corneal thickness increased and the injury of endothelium aggravated when intraocular tension increased. 　Conclusions:The results of ultrastructural examination indicated that destruction of the corneal endothelium following injury seems to be a consequential sequel. Dynamic corneal thickness measurement contributes to the judgement of degree of corneal endothelium injury. Control of intraocular tension is one of the important methods for preventing secondary injury of the corneal endothelium.

Objective To evaluate carotid intima-media thickness(IMT) and elasticity in patients with pregnancy-induced hypertension(PIH) using ultrasound radio-frequency data(RF-data) technology.Methods Twenty-seven PIH women(mean brachial blood pressure:108 mm Hg,1 mm Hg=0.133 kPa)were included and thirty age- and gestational week-matched normal pregnant women served as controls.Carotid IMT and stiffness were evaluated using quality IMT(QIMT) and quantitative artery stiffness(QAS)techniques with color Doppler.Results Carotid IMT and elasticity parameters,including pulse wave velocity,pressure at T1,arterial augmented pressure(AP) and arterial augmented pressure index(AIx) were significantly higher in PIH group than in the normal group[IMT:(466.84±118.50)μm vs (386.58±125.79)μm;PWV:(7.09±1.97)m/s vs (5.95±1.11) m/s;PT1:(127.50±14.29) mm Hg vs (105.89±11.02)mm Hg;AP:(5.14±3.39) mm Hg vs (1.98±2.19)mm Hg;AIx:(7.58±8.73)% vs (-4.79±7.92)%)], and there were significant differences(t=2.660,2.660,3.460,3.460,3.460,all P＜0.01).Conclusions PIH women have significantly increased carotid IMT and decreased elasticity compared with normal pregnant women.Radio-frequency technique could reliably reflect the changes of the carotid arterial structure and elasticity in patients with pregnancy-induced hypertension.

Objective To construct lentivirus containing CXCR4 gene and transfect MCF-7 cells , and obtain CXCR4 high-expressing MCF-7 cells. Methods CXCR4 gene was amplified by RT-PCR to construct CXCR4/pSin-EF2, which was transfected into HEK293T cells with psPAX2 and pMD2G vector for lentivirus packing. Packaged lentivirus was used to transfect human breast cancer cells MCF-7, with empty lentivirus as control. CXCR4 mRNA and protein expression levels were detected by RT-PCR and Western blot before and after transfection. And flow cytometry was used to detecte cell surface CXCR4 expression. Results The recombinant plasmid CXCR4/pSin-EF2 was constructed successfully,identified by double digestion and sequencing, and transfected into HEK293T cells to obtain high-titer lentivirus. RT-PCR and Western blot confirmed that the expression of CXCR4 in MCF-7 cells increased significantly after CXCR4 lentivirus transfection. Flow cytometry results showed that the CXCR4 positive rate increased from 26.78% to 99.29%, while there is no significant difference in CXCR4 expression between vector-transfected MCF-7 cells and non-transfected MCF-7 cells. Conclusion CXCR4 lentivirus and the breast cancer cell line with high and stable expression of CXCR4 (MCF-7CXCR4) were successfully constructed.

Objective To discuss the accuracy and objectivity of student standardized patients (SSP) in objective structured clinical examination(OSCE). Methods On March 30 and 31, 2013, 168 seven-year program medical students of class 2006 and 2007 took part in OSCE. Differences in as-sessment results between SPP and those given by professional doctors at 3 SP sites(angina pectoris SSP station, acute cholecystitis SSP station, depression SSP station) were analyzed. Each site had 4 items for assessment, with a total score of 100. Scores were given in strict accordance with a set of unified scoring rules. Counting data were presented as x±s. Data were verified using t test. P<0.05 was considered statis-tically considerable. Results At angina pectoris station, respective scores of SSP and professional physi-cians were 85.2±7.1 and 85.5±6.6, P=0.688. At acute cholecystitis station, respective score of SSP and professional physicians were 89.1±5.2 and 88.2±6.2, P=0.150. At depression station, respec-tive score of SSP and professional physicians were 79.8±7.5 and 78.2±7.0, P=0.078. Conclusion There is no statistical difference between scores given by SSP and those given by the physicians in OSCE. This proves that SSP who received standardized training delivers fair and accurate results in OSCE , and therefore is recommended for future application.

Utilizing standardized patient(SP) for classroom simulation is common in current medical education. In this paper, incentive measures and combination of SP with theoretical examination, simulated people and clinical practice were proposed after in-depth analysis of advantages and disadvan-tages of using medical students as SP in terms of recruitment, training, and application. All these mea-sures were intended to promote the development of simulative medical education that in turn to cultivate students to be competent in practice.

[Objective] To investigate the pathogen spectrum and characteristics of diarrheal disea-ses among children under five and to provide the basis for prevention and control measures . [ Methods] From February 2012 to January 2013, a total of 619 cases from 2 sentinel hospitals were collected for case survey .From them were collected stool specimens for detection of enteric pathogenic bacteria and virus nu-cleic acids. [Results] The virus detection rate was norovirus (11.95%), rotavirus(7.27%), enteric adenovirus(2.91%), saporovirus(1.45%) and astrovirus(0.97%).The bacteria detection rate was E. coli(5.65%), nontyphoidal salmonella (2.42%), campylobacter (0.81%), aeromonas hydrophila (0.65%),shigella(0.48%) and yersinia enterocolitica (0.32%).The virus detection rate was 27.70%from August to next March, which showed higher than that of other months (18.48%) (χ2 =7.18,P <0.05).The bacteria detection rate was 15.34%from April to October, showing higher than the rate of oth-er months(3.15%)(χ2 =31.22,P<0.05). [Conclusion] Norovirus is the main pathogen for viral diarrheal diseases in children under five and E .coli is the main pathogen of bacterial diarrheal diseases . All study results suggested that summer and autumn should be as the focus time in prevention and control of bacterial diarrhea , while those of viral diarrhea should cover the whole year .

To investigate the effect of recombinant human growth hormone (rhGH) on JAK2-STAT3 pathway and the growth of gastric cancer cell lines at different GHR expression status, the eukaryotic expression vector targeting human GHR (pGPU6/GFP/Neo-shGHR and pGPU6/GFP/Neo-scramble) was constructed and transfected into MGC803 cells by Lipofectamine 2000. Stable expressive cell lines were obtained by G418 screening. The expression of GHR was analyzed by Western blotting. After being stimulated with rhGH, cell growth was detected by MTT assay. Cell cycle and apoptosis were examined by flow cytometry. The components of JAK2/STAT3 signaling pathway were detected by Western blotting. There is no significant difference of GHR expression between MGC803 and pGPU6/GFP/Neo-scramble-transfected cells (named as MGC803-NC) (P > 0.05). Compared with MGC803, the GHR expression in pGPU6/GFP/Neo-shGHR-transfected cells (named as MGC803-shGHR) decreased significantly (protein decreased 50%). The cells were treated with rhGH at 0, 150 and 300 ng x mL(-1), the growth rate of MGC803 and MGC803-NC increased significantly, PI and the number of G2/M phase cells all increased significantly, and apoptosis decreased significantly. Western blotting revealed that the expression of pJAK2 and pSTAT3 was up-regulated after being treated with rhGH in MGC803 and MGC803-NC cells. In contrast, similar change was not observed in MGC803-shGHR cells. Knockdown of GHR gene may decrease the sensitivity of gastric cancer cells to rhGH, and down-regulating of components of the expression of JAK2/STAT3 signaling pathway may be the potential mechanisms.
Apoptosis ; Cell Cycle ; Cell Line, Tumor ; Cell Proliferation ; Down-Regulation ; Human Growth Hormone ; genetics ; pharmacology ; Humans ; Janus Kinase 2 ; metabolism ; RNA, Messenger ; metabolism ; RNA, Small Interfering ; genetics ; Receptors, Somatotropin ; genetics ; metabolism ; Recombinant Proteins ; genetics ; pharmacology ; STAT3 Transcription Factor ; metabolism ; Signal Transduction ; Stomach Neoplasms ; metabolism ; pathology ; Transfection

Animals ; Carps ; genetics ; Cloning, Molecular ; Fish Proteins ; genetics ; Hydroxylation ; Hypoxia-Inducible Factor 1, alpha Subunit ; genetics ; Lakes

OBJECTIVETo study the correlation between the inflammatory factors in the serum and the induced sputum and the hypothalamic-pituitary-adrenal (HPA) axis function in advanced lung adenocarcinoma patients of different syndromes.

METHODSTotally 71 patients with advanced lung adenocarcinoma were assigned to three groups according to syndrome differentiation, i.e., Shen-yang deficiency (SYD) group (28 cases), Fei-qi deficiency (FQD) group (23 cases), and yin deficiency fire excess (YDFE) group (20 cases). Another 41 healthy subjects were enrolled as the normal control group. Sputum was induced and blood samples were collected for measurement of cytokines including tumor necrosis factor alpha (TNF-alpha), transforming growth factor beta (TGF-beta), interleukin-6 (IL-6), and interferon gamma (INF-gamma). The cytokine assay was performed using Bio-Plex Pro multi assay technology. 24-h collection of urine was performed and salivary samples of the diurnal rhythm profiles [including urinary free cortisol (UFC), urinary 17-hydroxycorticosteroids (17-OH), urinary 17-ketosteroid (17-KS), and cortisol in the serum and saliva] were obtained for assessment of the HPA axis activity.

RESULTSA higher level of serum IL-6 and a lower level of 24-h UFC and 17-OH were found in the SYD group (P < 0.05). The urinary 17-KS was obviously lower in the SYD group than in the normal control group and the YDEE group (P < 0.05). Compared with the FQD group and the normal control group, a higher serum level of TNF-alpha and a lower level of IFN-gamma were found in the SYD group and the YDFE group (P < 0.05). The TNF-alpha and TGF-beta levels in the induced sputum obviously increased in the SYD group (P < 0.05). The IFN-gamma level in the induced sputum obviously decreased in the YDFE group (P < 0.05). The serum and salivary cortisol obviously decreased from 8: 00 am to 8:00 am the next morning in the SYD group (P < 0.05). The serum cortisol level was negatively correlated with serum TNF-alpha (r = -0.26, P = 0.03) and serum IL-6 (r = -0.25, P = 0.03). The salivary cortisol level was negatively correlated with IL-6 in the induced sputum (r = -0.29, P = 0.02). The serum IFN-gamma was positively correlated with urinary 17-OH (r = 0.21, P = 0.03).

CONCLUSIONSThe inflammatory factors of advanced lung adenocarcinoma patients of SYD syndrome were up-regulated, with the most obvious decreased or disarranged HPA axis functions. The levels of IL-6, TNF-alpha, and IFN-gamma were closely correlated with the HPA axis functions. The transformation from qi deficiency, yin deficiency to Shen-yang deficiency existed in lung adenocarcinoma patients. The levels of IL-6, TNF-alpha, and IFN-gamma in the serum and the induced sputum, as well as the HPA axis functions are important indices for microscopic syndrome typing of lung adenocarcinoma.

Adenocarcinoma ; blood ; physiopathology ; Adolescent ; Adult ; Aged ; Case-Control Studies ; Cytokines ; blood ; Female ; Humans ; Hypothalamo-Hypophyseal System ; metabolism ; physiopathology ; Inflammation ; Lung Neoplasms ; blood ; physiopathology ; Male ; Middle Aged ; Neoplasm Staging ; Pituitary-Adrenal System ; metabolism ; physiopathology ; Sputum ; chemistry ; Young Adult

OBJECTIVETo explore the biomarkers and inflammatory characteristics for microcosmic syndrome differentiation of cold-phlegm syndrome (CPS) and heat-phlegm syndrome (HPS) in patients with bronchial asthma.

METHODSPatients with bronchial asthma of chronic persistent condition were distributed into three groups according syndrome differentiation, the CPS group (27 patients), the HPS group (32 patients) and the non-cold/heat-phlegm syndrome group (NP group, 31 patients), besides, a control group was setup with 33 healthy persons. Percentages of neutrophils and eosinophils (NEU, EOS) in sputum sample (collected by induction) and peripheral blood were counted; and levels of interleukin-8, -5, and -4 (IL-8, IL-5 and IL-4), interferon-gamma (IFN-gamma), leukotriene B4 (LT-B4), eosinophil cationic protein (ECP), and C-reactive protein (CRP) in sputum supernatant and serum were determined by enzyme-linked immunosorbent assay.

RESULTSPercentage of NEU in sputum of HPS group was higher than that in the other three groups (P < 0.05); while percentages of EOS in serum and sputum of CPS group were higher than that in the other three groups (all P < 0.01). Level of ECP (a parameter closely associated with EOS) also was high in the CPS group, but IL-8 (a parameter closely associated with NEU) showed no significant difference in various groups (P > 0.05). Moreover, the CPS group showed a higher serum IL-4 (P < 0.05) but a lower IFN-gamma/IL-4 level as compared with those in the NP group (P < 0.01).

CONCLUSIONPhlegm, which is considered by Chinese medicine as an inveterate root of asthma, might be closely related with the inflammation in modern medicine. The inflammatory characteristics of asthma in patients with CPS partially present as increase of EOS, possibly show Th2 dominant trend, similar to that presented in eosinophilic asthma. Asthma with HPS embodies increase of NEU in respiratory tract. EOS and ECP might be the important markers for microcosmic syndrome differentiation of CPS, and NEU might be that for HPS.

Adolescent ; Adult ; Aged ; Asthma ; complications ; diagnosis ; pathology ; Biomarkers ; metabolism ; Case-Control Studies ; Diagnosis, Differential ; Eosinophil Cationic Protein ; metabolism ; Eosinophils ; pathology ; Female ; Humans ; Inflammation ; complications ; diagnosis ; pathology ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Neutrophils ; pathology ; Young Adult