Objective To study the receptors for the axon guidance factor Netrin-4 and whether the known Netrin-1 receptors, DCC and UNC5H1, are coreceptors for Netrin-4. Methods The receptor for Netrin-4 was detected by affinitycytochemistry and in situ staining with the alkaline phosphatase (AP) tagged Netrin-4 fusion protein. The dynamic character of the binding between Netrin-4 and its receptor was evaluated through ligand-receptor binding assay and competitive binding assay. Results In situ staining of AP4-Netrin-4 protein bound to COS7 cells transiently transfected with eukaryotic expression coding DCC and UNC5H1 showed that the Netrin-4 could bind to the transfected cells. Competitive binding assays showed that Netrin-1 could compete with Netrin-4 for the same receptors. Conclusion DCC and UNC5H1 are coreceptors for Netrin-4. However, because the binding affinity of Netrin-4 with its receptor was lower than that of Netrin-1,it could be inferred that there should exist unique receptor(s) for Netrin-4 other than DCC and UNC5H1.

Objective To explore the efficacy and adverse reaction of nedaplatin (NDP)+gemcitabine (GEM)and cisplatin (DDP)+GEM for advanced lung squamous cell carcinoma.Methods A total of 1 01 cases advanced untreated patients from September 201 2 to December 201 3 were randomly divided into 2 groups using random number table method:69 patients in the observation group accepted NDP+GEM treatment and 32 patients in the control group received DDP +GEMtreatment.The objective response rate (RR),disease control rate (DCR ) and progression-free survival (PFS ) and adverse reaction were collected and evaluated. Results RR was 28.6%(1 8/63)in the observation group and 1 5.6%(5/32)in the control group,DCR was 81 .0%(51/63)in the observation group and 68.8%(22/32)in the control group (χ2 =1 .36,P=0.24;χ2 =1 .67,P=0.20).The median PFS was 4.52 months and 4.01 months in the observation group and control group (χ2 =0.09,P=0.73).The major adverse reaction was myelosuppression in both groups (33.3% vs.37.5%,χ2 =0.1 7,P=0.68).The incidence ofⅢ-Ⅳ grade nausea and vomiting was lower in the observation group, compared with the control group (1 4.5%vs.56.3%,χ2 =1 9.02,P=0.05).Conclusion NDP combined with GEM in advanced lung squamous cell carcinoma of the first-line treatment has equivalent efficacy to DDP+GEM, with lower incidence of adverse reaction,which is worthy of further dissemination of research.

Objective　To explore the role of tyrosine hydroxy lase (TH) gene in the treatment of Parkinson's disease (PD) by using ex vivo gene transfer. Methods　After the construction of TH gene in a retroviral vector, the astrocytes were cultured with the supernatant containing the recombinant DNA and then grafted into the cerebrum of PD rats. The reduction of the rat rotation beharior was evaluated. Results　The rotati on of PD rats was markedly improved in the rats with ex vivo gene transfer. Conclusion　TH has an obvious efficiency on the treatment of PD and the astrocytes can be used as effective gene transfer cells.

AIM To study the chemical constituents from the leaves of Cyclocarya paliurus (Batal.) lljinskaja.METHODS The ethyl acetate fraction of methanol extract from C.paliurus leaves was isolated and purified by silica,ODS and semi-preparative HPLC,then the structures of obtained compounds were identified by spectral data.RESULTS Eleven compounds were isolated and identified as (6S,7R,8R)-7α-[(β-glucopyranosyl) oxy] lyoniresinol (1),lyoniside (2),(-)-lyoniresinol 3 α-O-β-D-xylopyranoside (3),dihydrodehydrodiconiferyl alcohol 4'-O-β-D-glucoside (4),kaempferol-3-O-α-L-rhamnoside (5),kaempferol-3-O-α-L-(4"-Z-p-cumaroyl)-rhamnoside (6),naringenin (7),4'-hydroxywogonin (8),1-(3',4'-dihydroxyphenyl)-7-(4"-hydroxyphenyl)-4-hepten-3-one (9),grasshopper ketone (10),p-hydroxybenzoic acid (11).CONCLUSION Except for compounds 5,11,all the compounds are isolated from this plant for the first time.

Objective To validate the four chronic kidney disease epidemiology collaboration (CKD-EPI) predictive equations based on serum creatinine (SCr) and cystatin C (Cys C) in Chinese CKD patients,and try to develop the GFR predictive equations for Chinese CKD patients.Methods254 CKD patients were randomly selected from four Grade ⅢA hospitals in different regions in China from September 2007 to December 2010.Clearance of dual plasma sampling 99mTc-DTPA was used to measure glomerular filtration rate (rGFR) in 254 CKD patients.The serum concentration of Cr and Cys C were measured.CKD-EPI SCr equation,Cys C equation,Cys C equation adjusted for age,sex and race,SCr/Cys C combinated equation adjusted for age,sex and race were used to estimate GRF ( labeled as eGFR1,eGFR2,eGFR3 and eGFR4,respectively).The correlation,bias and precision of eGFRs were compared with rGFR by Wilcoxon signed rank test,intraclass correlation coefficient (ICC) and Spearman correlation analysis.The deviation degree between rGFR and different eGFRs was compared via Bland-Altman graph.The accuracy within 15％,25％,30％ ( P15,P25,P30) and the staging correctness of eGFR against CKD at different stages was calculated.ResultsThe rGFR in 254 CKD participants was [ 48.07 (26.19 -92.97 )] ml · min -1·(1.73 m2) -1.The Spearman correlatiou coefficients (CC) of eGFR and rGFR varied within the range of 0.873 - 0.896 ( P =0.000 ).The intra-class CC ( ICC ) varied within the range of 0.920 - 0.942.The correlation of eGFR4 was the best.The absolute deviations of 4 eGFRs and deviation precision were eGFR4 ＜eGFR3 ＜ eGFR2 ＜ eGFR1.The 95％ confidence intervals for the regression line of 4 eGFRs shown by Bland-Altman graphs were 92.5,87.3,83.0 and 76.1 ml · min-1 · ( 1.73 m2 ) -1,respectively,with the best result of eGFR4.For P30,the correctness of 4 eGFRs were eGFR4 ＞ eGFR3 ＞ eGFR2 ＞ eGFR1,but no significant difference was found by Chi square test (x2 =6.448,P =0.092).The overall correctness rate in 4 eGFRs against CKD stages were 48.4％ -57.5％,with the highest consistency of eGFR4,but their staging correctnessratewerenotideal(Kappa values were 0.405,0.348,0.366 and 0.463,respectively).Conclusions Compared with CKD-EPI SCr equation,no advantage was found in CKD-EPI Cys C equation.The Cys C equation adjusted by age and sex shows a little advantages over CKD-EPI Cys C equation in bias,precision,correlation and accuracy.The CKD-EPI SCr/Cys C combinated equation adjusted by age,sex and race has advantage over other three equations not only in bias,precision,correlation and accuracy,but also in staging correctness.However,the validation of this equation is still not fairly ideal for Chinese CKD patients.Based on these findings,it is essential for the Chinese CKD patients to develop SCr/Cys C combined predictive equation which adjusted by age,sex or other factors.(Chin J Lab Med,2012,35:798-804)

Adolescent ; Adult ; Cardiac Catheterization ; economics ; methods ; Cardiac Surgical Procedures ; economics ; methods ; Cardiac Valve Annuloplasty ; Child ; China ; epidemiology ; Echocardiography ; Female ; Heart Septal Defects, Ventricular ; complications ; diagnostic imaging ; surgery ; Humans ; Length of Stay ; statistics & numerical data ; Male ; Operative Time ; Postoperative Complications ; epidemiology ; Septal Occluder Device ; Tricuspid Valve Insufficiency ; complications ; diagnostic imaging ; surgery ; Young Adult

OBJECTIVETo observe the temperature change of traditional silver needle in the human body during the burning of moxa ball.

METHODSThirty-six healthy volunteers were randomly divided into a single-needle group and a multi-needle group, 18 cases in each group. For both groups, one silver needle (18 cm in length, 1.1 mm in diameter), which was adopted in this research to measure the temperature change, was punctured in the insertion point of the volunteer (inside the top of the left buttock, 7 cm under the edge of the highest point of the iliac crest, 7 cm lateral to the dorsomedian line), then another four silver needles were punctured 2 cm respectively anterior, posterior and lateral to the insertion point in the multi-needle group, and all the silver needles were inserted with 6 cm depth. Afterigniting the 1.3 g moxa ball on the needle tail, the temperature of the measuring points that were 3 mm, 33 mm, and 63 mm above the silver needle tip were recorded separately by digital temperature measuring instrument.

RESULTSThe peak temperature of the three measuring points in the single-needle group was all around 41 degrees C, while those in the multi-needle group were around 43 degrees C, which had significant differences (all P < 0.05), but no significant differences among the highest temperature of the measuring points in the same group could be found (all P > 0.05). The highest temperature of moxa ball in the single needle group was (611.16 +/- 6.91) degrees C, while that of the central moxa ball in the multi-needle group was (628.94 +/- 8.99) degrees C, the difference of which was significant difference (P < 0.01).

CONCLUSIONThe temperature conductivity of the silver needle is very well, so the heat of the moxa ball could pass from the tail of needle to the tip during the warming treatment. The peak temperature on the body, tip of the silver needle in the multi-needle group is higher than those in the single needle group. Also, the peak temperature of multi-moxa ball is higher than that of single moxa ball.

Acupuncture Therapy ; instrumentation ; Adult ; Female ; Hot Temperature ; Humans ; Male ; Middle Aged ; Needles ; Young Adult

In vitro method as indispensable biological research and testing tools, has been widely used in cosmetics compliance test, biological mechanism research and functional materials screening. These methods include alternative methods which have been accepted by reguations and standardized test guide, non-testing method used for risk assessment and prediction, and a lot more diversification and individualization of in vitro methods. With the consensus formation of the whole industry and increase of application experience, innovation and optimization in vitro method will be constantly emerge. These technologies will be beneficial to improve competitiveness and to develop products to meet the needs of consumers.

miR-214 plays a major role in the self-renewal of skin tissue. However, whether miR-214 regulates the proliferation and differentiation of human hair follicle stem cells (HFSCs) is unknown. Primary HFSCs were isolated from human scalp skin tissue, cultured, and identified using flow cytometry. An miR-214 mimic and inhibitor were constructed for transfection into HFSCs. The MTS and colony formation assays examined cell proliferation. Immunofluorescence detected the localization and expression levels of TCF4, β-catenin, and differentiation markers. Luciferase reporter and TOP/FOP Flash assays investigated whether miR-214 targeted EZH2 and regulated the Wnt/β-catenin signaling pathway. Western blot determined the expression levels of enhancer of zeste homolog 2 (EZH2), Wnt/β-catenin signaling-related proteins, and HFSC differentiation markers in cells subjected to miR-214 transfection. miR-214 expression was remarkably decreased during the proliferation and differentiation of HFSCs into transit-amplifying (TA) cells. Downregulation of miR-214 promotes the proliferation and differentiation of HFSCs. Overexpression of miR-214 led to decreased expression of EZH2, β-catenin, and TCF-4, whereas downregulation of miR-214 resulted in increased expression of EZH2, β-catenin, and TCF-4 as well as TA differentiation markers. Immunofluorescence assay revealed that inhibiting miR-214 triggered the entry of β-catenin and TCF-4 into the nucleus. The luciferase reporter and TOP/FOP Flash assays demonstrated that miR-214 directly targets EZH2 and affects Wnt/β-catenin signaling. The miR-214/EZH2/β-catenin axis could be considered a candidate target in tissue engineering and regenerative medicine for HFSCs.
Antigens, Differentiation ; Blotting, Western ; Cell Proliferation ; Down-Regulation ; Flow Cytometry ; Fluorescent Antibody Technique ; Hair Follicle* ; Hair* ; Humans* ; In Vitro Techniques* ; Luciferases ; Regenerative Medicine ; Scalp ; Skin ; Stem Cells* ; Tissue Engineering ; Transfection

Objective To study the changes of substance P (SP) content in plasma of asthmatic children and in theblood, bronchoalveolar lavage fluid (BALF) and lung tissues of asthmatic guinea pigs and SP mRNA expression, and toinvestigate the relationship between SP and asthma of children. Methods We determined the contents of plasma SP bythe method of immunoassay at the stage of acute attack and clinical remission in 35 children with asthma. On the basis ofasthma model of guinea pigs, we examined the contents of SP in the blood, BALF and lungs. The expression of Spthat of the stage of clinical remission and normal controls ( P ＜0.01). At the stage of clinical remission, the differencethe blood, BALF and lungs were higher than those of normal ones ( P ＜0.01), and the expressions of SP mRNA inlungs were higher ( P ＜0.01). In repeatedly attack group, the SP content of guinea pigs was higher than that of asthmagroup ( P ＜ 0.05), and the expression of SP mRNA in lungs was also higher ( P ＜ 0.01 ). Conclusions SP levelsincreased in asthmatic children and asthmatic guinea pigs, and the expression of SP mRNA was up-regulated in the lungsof asthmatic guinea pigs. They were positive related to the severity of asthma. So SP has close relationship with asthmaand may play a role in the mechanism of asthma.