Our aim in this study was to determine differentially expressed genes of CD34 + cell from bone marrow and G CSF mobilized peripheral blood. CD34 + cells isolated from bone marrow and G CSF mobilized peripheral blood of the same donor were identified for differentially expressed genes in CD34 + cells using the technique of suppression subtractive hybridization. Eleven genes were expressed with higher levels in CD34 + cells from mobilized peripheral blood, as compared with data of GenBank. These genes included nuclear proteins, transcriptional regulatory molecules, zinc finger proteins and interferon induced molecules. These results demonstrate that there are some differences between the two groups of CD34 + cells. Further study would give us more extensive understanding about mobilization and homing of hematopoietic stem cells.

Objective: To investigate the expression of MDR1 and GFP in the human hematopoietic cells mediated by adeno-associated virus. Methods: The GFP gene was transferred into the human hematopoietic cells by AAV vectors and created strong visible fluorescence by purely molecular biological means. Using adeno-associated virus vectors, we have transferred human mdr-1 gene into human hematopoietic cells and investigated the drug resistence of human hematopoietic cells modified with mdr-1 gene. PCR analysis confirmed that mdrl cDNA had been successfully transferred into the human hematopoietic cells. An assay of MTT proved that the human hematopoietic cells modified by mdrl gene had resistance to colchicine. Results: It was about 30% of the hematopoietic cells that expressed the green fluorescent proteins. The resistance of hematopoietic cells was increased parently when the cells were infected by the crude virus stocks. Conclusion: It is conducted that the AAV vector could successfully transfer the foreign gene into the human hematopoietic cells. The cells modified with mdrl gene have increased the resistance to drugs.

Objective To explore the possible risk factors for autism spectrum disorders (ASD),and to pro-vide a basis for exploring the etiology of the disease.Methods This case -control study included 68 patients diag-nosed as ASD for a first lifetime(according to the fourth edition of the American Diagnostic and Statistical Manual of Mental Disordersfor ASD diagnosis)from May 201 4 to January 201 5,and 77 non -ASD controls (normal children, matched on gender)in Jiangxi Children′s Hospital were selected to undergo the risk factor survey for ASD.The survey content included 1 0 categories:general status,birth,feeding,the past history,mother′s pregnancy and her health condi-tion during pregnancy and environmental exposure,parents′occupational exposure,family history and relevant test re-sults.Logistic regression analysis was performed to analyze the results of the survey.Results The possible risk factors for ASD increased if mother had virus infection 2 years before pregnant (OR =7.97,95%CI:2.42 -26.31 ),had occu-pational exposure (OR =3.99,95%CI:1 .27 -1 2.52),volatile organic compounds exposure during pregnancy (OR =22.21 ,95%CI:2.28 -21 6.09),as well as living closely to transport passage ways during pregnancy (OR =0.59,95%CI:0.38 -0.93)or having a family heredity history (OR =58.50,95%CI:5.81 -589.57).Breastfeeding (OR =0.81 ,95%CI:0.66 -0.98)might be a protective factor in ASD.Conclusions In addition to genetic factors,the ute-rine environment from conception to birth and growth environment play an important role in the pathogenesis of ASD.

Objective:To clone and express human Fas activation domain(FasAD)with the bioactivity.Methods:The FasAD cDNA was amplified by seminested RT PCR,and then inserted into the prokaryote vector of pTYB12 expressed intein protein to construct the recombinant plasmid of FasAD pTYB12.The FasAD peptide was expressed and purified in IMPACT TM CN system by the method of one step affinity purification.Results:Sequence analysis revealed that cloned FasAD cDNA sequence was completely same as that of Genebank record(M67454).Soluble fusion protein was successfully expressed by induction of IPTG.The FasAD peptide with a molecular weight of 5 000 was obtained by purification and was recognized by the rabbit anti human Fas polyclonal antibody in Western blot analysis.It’s activity of inhibition of apoptosis induced by rhFasL can each to 70% in primary biological detection.Conclusion:The above results indicated that FasAD peptide could be prepared by using the IMPACT TM CN system,thus laid a relatively experimental foundation for further research of relationship between structure, function and interaction with it’s ligands,and for further development of biological immune modulator. [

BACKGROUND: Rabbit serum samples are widely used in basic researches, and two-dimensional gelelectrophoresis (2-DE) is the most classic technique for protein separation. Therefore, it is of great significance to establish a stable technique system of 2-DE for rabbit serum.OBJECTIVE: To establish a 2-DE technique system for rabbit serum protein separation. DESIGN, TIME AND SETTING: A single sample observational experiment was performed at Tianjin Key Laboratory of Biomarkers for Occupational and Environmental Hazard of Chinese People's Armed Police Forces Medical College from June to July in 2008. MATERIALS: Six healthy rabbits were provided by the Animal Experimental Center of Tangshan Vocational Technical College METHODS: Health rabbit serum was dissolved in rehydration sample loading buffer before and after eliminating high abundance proteins to make proteins in it schizolysis adequately. After reductive alkylation, the samples were loaded into the rehydration tray to undergo passive rehydration for 14 hours. Isoelectric focusing (IEF) electrophoresis was followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). After sliver staining, the gel was analyzed by PDQuest7.4. MAIN OUTCOME MEASURES: ①Efficiency of eliminating high abundance proteins. ②Two-dimensional eleotrophoregrams (2-D electrophoregrams)RESULTS: Distinct 2-D eleotrophoregrems were obtained with high resolution and good reproducibility. The removal of high abundance proteins in serum failed to result in better 2-D electrophoregrams.CONCLUSION: We have successfully established a 2-DE technique for rabbit serum proteome, which can lay the foundation for the further study of serum proteomics of diseases.

Objective To investigate the effects of post-operative analgesia with oxycodone or morphine for patients undergoing colon cancer radical surgery on platelet activation and cellular immunity.Methods Forty colon cancer patients scheduled for radical surgery, 23 males and 17 females, ASA physical status Ⅰ or Ⅱ, were randomly divided into 2 groups (n=20 each): oxycodone group (group O) and morphine group (group M).Patient-controlled intravenous analgesia (PCIA) was used for post-operative analgesia.PCIA solution contained oxycodone 1 mg/kg and tropisetron 6 mg in 100 ml normal saline in group O or morphine 1 mg/kg and tropisetron 6 mg in 100 ml normal saline in group M.Blood samples were obtained from the patients at 5 min before anesthesia induction (T0), 4 h after surgery (T1), 24 h after surgery (T2) and 48 h after surgery (T3).The levels of glycoprotein (GP)Ⅱb/Ⅲa, P-selection (CD62P), natural killer (NK) cells, NKT cells, and natural Treg (nTreg) cells were detected.The platelet aggregation rate (PAR) was determined.Results Compared with T0, the levers of GPⅡb/Ⅲa, CD62P, PAR and nTreg cells were significantly higher at T1 in group O and at T1, T2 in group M (P<0.05).Compared with T0, the levels of NK and NKT cells were decreased significantly at T1 in group O and at T1-T3 in group M (P<0.05).The levels of GPⅡb/Ⅲa, CD62P, PAR and nTreg cells at T2 and T3 in group O were decreased significantly as compared with group M (P<0.05).The levels of NK cells, NKT cells at T2 and T3 in group O were significantly higher than those in group M.Conclusion Post-operative analgesia with oxycodone for patients undergoing colon cancer radical surgery exhibits a more significant effect of decreasing platelets activity and presents a less disturbance on cellular immunity as compared with morphine.

Objective To analyze the value of the potential risk factors on predicting primary graft dysfunction (PGD) after bilateral lung transplantation for the patients with idiopathic pulmonary fibrosis (IPF).Methods A retrospective study was conducted. Fifty-eight patients with IPF who underwent the bilateral lung transplantation admitted to Wuxi People's Hospital Affiliated to Nanjing Medical University from June 2014 to March 2017 were enrolled. The grade 3 PGD happened within 72 hours after transplantation was taken as the outcome event, and these patients were divided into PGD and non-PGD groups. The age, gender, body mass index (BMI), underlying disease, and N-terminal-probrain natriuretic peptide (NT-proBNP) before operation, pulmonary artery systolic pressure (PASP), pulmonary artery diastolic pressure (PADP), and mean pulmonary artery pressure (mPAP) before and after operation, duration of operation, the volume of blood transfusion during operation and postoperation, the use of extracorporeal membrane oxygenation (ECMO) during the operation, blood purification treatment after operation, and shock within 3 days after operation were recorded. The differences of parameters mentioned above between the two groups were compared. The predictive factors of PGD were searched by binary logistic regression analysis, and the receiver operating characteristic curve (ROC) was plotted to analyze the predictive value of preoperative PADP for grade 3 PGD after transplantation.Results Among 58 patients who underwent the bilateral lung transplantation, 52 patients were enrolled. The rest patients were excluded because of incomplete clinical data. There were 17 patients in the PGDgroup, with a mortality rate of 47.06%. The non-PGD group included 35 patients with a mortality rate of 8.57%. PADP and mPAP ahead of operation, the dosage of red cells suspension after the operation, and the total amount of blood transfusion during and after the operation in PGD group were significantly higher than those in non-PGD group [PADP ahead of operation (mmHg, 1 mmHg = 0.133 kPa): 33.7±10.5 vs. 25.3±10.1, mPAP ahead of operation (mmHg): 40.4±14.1 vs. 32.8±11.1, the dosage of red cells suspension after the operation (mL): 700 (300, 1500) vs. 300 (300, 500), the total amount of blood transfusion during and after the operation (mL): 2250 (1850, 4275) vs. 1800 (1550, 2800)], with statistically significant differences (all P < 0.05). There were no significant differences in age, gender, BMI, underlying disease, NT-proBNP before operation, PASP before and after operation, PADP and mPAP after operation, duration of operation, amount of plasma and red cells suspension as well as total amount of blood transfusion during operation, plasma amount and total amount of blood transfusion after operation, amount of plasma and red cells suspension during and after operation, use of ECMO during operation, blood purification treatment after operation, and shock after operation between the two groups (all P > 0.05). It was shown by binary logistic regression analysis that the preoperative PADP was the independent risk factor of grade 3 PGD after lung transplantation [odds ratio (OR) = 1.084, 95% confidence interval (95%CI) = 1.016-1.156,P = 0.015]. It was shown by ROC curve that the area under the ROC curve (AUC) of the PADP before operation for predicting the grade 3 PGD after lung transplantation was 0.728. When the cut-off value was 36 mmHg, the sensitivity was 47.1%, and the specificity was 91.4%.Conclusions Compared with the non-PGD group, the patients with higher preoperative PADP were more common in the PGD group, and the patients in the PGD group were more likely to be characterized by grade 3 PGD after lung transplantation. The preoperative PADP was an effective predictor of grade 3 PGD after lung transplantation.

OBJECTIVETo perfect the current standard of Rhizoma Diosoreae and Rhizoma Diosoreae stir-baked with bran by improving quality standards of the two processed pieces.

METHODThe quality standards were established according to 9 batches of processed pieces, separately. The standards contains items of identification, water, total ash, acid-insoluble ash, extractives, heavy metals limit, organochlorine limit, microbial limit and assay.

RESULTThe TLC of the two pieces was characteristed. The contents of acid-insoluble ash in the two pieces were increased, not more than 0.5%, 0.3%, respectively. The content limits of five kinds of heavy metals and harmful elements, two kinds of residual organochlorine pesticides and three microbial limits were increased. There were no more than 2 x 10(-7) of lead, 2 x 10(-7) of cadmium, 1 x 10(-5) of copper, 3 x 10(-7) of arsenic, 1 x 10(-7) of mercury, 1 x 10(-7) of hexachlorocyclohexane (BHC) and 1 x 10(-7) of chlorophenothane (DDT) in the two processed pieces, respectively. There were no more than 2 000 and 600 cfu x mL(-1) in the two pieces, respectively and no more than 30 MPN x 100 g(-1) and fungi can not be tested in the two pieces. The contents of allantoin in the two pieces were no more than 0.15%.

CONCLUSIONThis method is simple and suitable for the quality control of the two processed pieces.

Chemistry, Pharmaceutical ; methods ; standards ; Chromatography, Thin Layer ; Dioscorea ; chemistry ; Quality Control ; Rhizome ; chemistry

OBJECTIVETo study the processing technics of prepared slice of Rhizoma Dioscoreae for its industrial production.

METHODThe effect factors, such as stiring temperature (60, 80, 100 degrees C), stiring time (10, 20, 30 min) and the proportion of pieces and wheat bran (100: 5, 100: 10, 100: 15) were evaluated by orthogonal method. The content of allantoin in Rhizoma Dioscoreae was determine by HPLC method.

RESULTThe amount of wheat bran could affect the content of allation significantly, and stiring temperature and stiring time almost had no effect on content of allantoin. The processing technics was tested by industrial produce, according to 11 batches from 4 growing regions. The result showed that the quality of industrial product was stable.

CONCLUSIONThat the amount of wheat bran can affect the quality of Rhizoma Dioscoreae greatly. The determined technics is simple and suitable for prepared slice of Rhizoma Dioscoreae industrival manufacture.

Allantoin ; analysis ; Dietary Fiber ; analysis ; Drug Compounding ; methods ; Drugs, Chinese Herbal ; chemistry ; Pinellia ; chemistry ; Technology, Pharmaceutical

Objective To investigate the evaluation index of melasma staging by clinical manifestations and non-invasive skin detection technology.Methods A total of 195 patients with a clinical diagnosis of melasma were enrolled from the First Affiliated Hospital of Kunming Medical University.The skin with lesion enlarged,color darker,erythema,red occured after scratching or lesion faded after compressing with glass belonged to the active stage;on the contrary,it was in the stable stage.Reflectance confocal microscopy (RCM),dermoscopy,Mexameter 18 and LAB were used to observe skin lesions of different stage of melasma.Results There were 115 patients (59.0 ％) in the active stage of melasma and 80 patients (41.0 ％) in the stable stage.DMA score in active stage 35.08± 10.59 were significantly higher than that of the stable stage 15.06-4-9.20 (P＜0.05).There were statistically significant difference in the quantity of inflammatory cell and blood vessels between two stages of melasma (P＜0.05).Erythema index (EI) in active stage of melasma 376.35±61.39 were high-er than that of the stable stage 320.364± 62.40 (P＜0.05).A-value in active stage of melasma 13.28± 1.75 were higher than that of the stable stage 12.34± 1.78 (P＜0.05).However,there were no siginificant differences in the quantity of melenin,melanin index (MI),L-value and B-value.Conclusions Melasma could be divided into active stage or stable stage,respectively,according to its clinical manifestations.DMA score,quantity of inflammatory cells and blood vessels,EI and A-value could be used as the reference index of melasma staging.