BACKGROUND:Large-scale expansion of undifferentiated and multipotential adipose-derived stem cells using serum-free culture system is a difficult issue to be resolved. OBJECTIVE:To establish an in vitro culture system combined with the extracellular matrix in order to investigate the efficiency, effectiveness and security of extracellular matrix on expanding adipose-derived stem cells. METHODS:In vitro isolated adipose-derived stem cells were seeded in traditional two-dimensional plastic plates and extracellular matrix-coated plates supplemented with serum-free medium respectively. After in vitro expansion, total cellnumber, expression of cellsurface markers, cellsenescence degree and multipotent differentiation ability (adipogenic, osteoblastic and chondrogenic differentiation) of adipose-derived stem cells cultured under both conditions were detected and compared. Moreover, the clinical safety of adipose-derived stem cells expanded in extracellular matrix-coated plates was investigated. RESULTS AND CONCLUSION:Total cellnumber of passage 5 adipose-derived stem cells cultured in extracellular matrix-coated plates was 10 times more than that in traditional two-dimensional plastic plates. Flow-cytometric analysis showed that adipose-derived stem cells cultured with extracellular matrix expressed stem cellsurface markers. cellular senescence examination showed that almost al of passage 15 adipose-derived stem cells cultured with extracellular matrix showed no aging, while most passage 5 adipose-derived stem cells cultured by the two-dimensional system aged and lost their proliferation ability. Multidirectional induction of adipose-derived stem cells showed that passage 15 adipose-derived stem cells cultured with extracellular matrix could stil differentiate into adipocytes, osteoblasts and chondrocytes as passage 5 adipose-derived stem cells did, which performed much better than the induced differentiations of passage 5 adipose-derived stem cells cultured by the two-dimensional system. Karyotype analysis and in vivo invasion experiment insured the clinical safety of adipose-derived stem cells expanded with extracellular matrix. Al above results suggest a safe and more efficient expansion system of extracellular matrix for clinical application using the serum-free culture system combined with extracellular matrix.

Objective To investigate the epidemiological characteristics of incipient neonatal hyperbilirubinemia. Methods The clinical data of admitted neonates with hyperbilirubinemia were retrospectively analyzed from June 2012 to May 2013. Results Two hundred and eight-four neonates with hyperbilirubinemia were enrolled and the ratio of male:female was 1.51:1. For the causes of hyperbilirubinemia, the incidences of ABO hemolytic and sepsis were higher in term infants than those in preterm infants, and the incidences of pneumonia, necrotizing enterocolitis and intracranial hemorrhage were higher in preterm infants than those in term infants (P<0.05). Compared with the preterm infants, the term infants had jaundice appearance and peak at earlier time, shorter duration of jaundice, faster decline rate of jaundice, higher levels of albumin and indirect bilirubin at the peak of jaundice (P<0.01). In the term infants, the time of jaundice appearance and peak were earlier in hemolytic group than those in non-hemolytic group (P<0.05). In preterm infants, the peak of transcutaneous bilirubin was higher in hemolytic group than that in non-hemolytic group (P<0.05). Six cases with bilirubin encephalopathy had abnormalities cranial MRI imaging, and the MRI was not entirely consistent with the peak level of bilirubin. Conclusions There are clinical differences between hemolytic and non-hemolytic hyperbilirubinemia in both term and preterm infants.

Objective To investigate the effect of intra-articular carboxymethylated ehitosan(CM- CTS)injection on inducible nitric oxide synthase(iNOS)expression in cartilage at the early stage of os- teoarthfitis(OA).Methods Thirty-two white rabbits were underwent unilateral anterior cruciate ligament transection(ACLT)and were randomly divided into 4 groups 5 weeks after transection.Rabbits of group A re- ceived 0.3 ml of 2% high molecular weight CMCTS(H-CMCTS)once every two weeks.Rabbits in group B were treated using 2% low molecular weight(L-CMCTS)CMCTS at:the same intervals.Group C rabbits were injected intra-articularly with 0.3 ml of 1% sodium hyaluronate(Na-HA)once a week.Animals of group D were not injected.At sacrifice,11 weeks following surgery,the expression of iNOS in cartilages was analyzed by immunohistochemistry and reverse transcription-polymerase chain reaction(RT-PCR)methods.Results Both immunohistochemistry and RT-PCR showed that the level of iNOS expression of cartilage in CMCTS in- jection groups was lower than that in Na-HA injection group and the untreated group.There was no significant difference in iNOS expression between the two different molecular weight CMCTS injection groups. No signifi- cant difference of iNOS expression in cartilage was found between Na-HA injection group and the untreated group.Conclusion CMCTS suppresses iNOS expression in cartilage during the early stage of OA.Na-HA treatment has no effect on iNOS expression in cartilage.

Objective To evaluate the effectiveness and safety of gefitinib retreatment beyond progression(GRBP)in non-small cell lung cancer(NSCLC)patients with rare EGFR mutations.Methods We retrospectively analyzed six rare-EGFR-mutation NSCLC patients from Jan 2011 to Dec 2015.Those patients had previous disease control and then disease progression according to Response Evaluation Criteria in Solid Tumors version 1.1(RECIST v1.1)after taking oral gefitinib 250 mg once a day.After that,continuing gefitinib was decided by clinicians′ experience at the same treatment option.The primary endpoints were response rate(RR),overall survival(OS),the first and second progression-free survival(PFS-1 and PFS-2).Safety was assessed according to the NCI-CTCAE version 4.0.Results After initial treatment of gefitinib,4 patients achieved partial response(PR)and 2 patients showed stable disease(SD),with RR being 66.7％.The median PFS-1 and PFS-2 were 10 months(95％CI 6.6-13.4)and 9 months(95％CI 6.9-11.1),respectively.The median OS time was 28 months(95％CI 10.4-45.6).The most common treatment-related adverse events were fatigue,diarrhea,rash,itching and elevated transaminases.Conclusion In our study,gefitinib retreatment beyond disease progression is effective with a manageable tolerability profile.

Objective Toinvestigate the association between genetic susceptibility of gastric carcinoma and single nucleotide polymorphism of REG1a gene in Chinese Han population. Methods The genotype of REG1a gene were detected by PCR and DNA sequencing in 183 Chinese patients with gastric carcinoma and 204 controls.Results It was found 3 new SNPs in REG1a gene at site 929(T/C),1790(C/G) and 2751(A/T), respectively; and the different genotypes of site 929 and 1790 were significant between gastric carcinoma patients and controls(P<0.05). Conclusions Sequencing of REG1a gene from specific population is available for screening candidate SNPs. The study reveals that the initially gastric carcinoma may be associated with site 929 and 1790. It provides a basis for further screening suitable SNPs labeling forecasting risk for gastric carcinoma.

Objective To investigate the relationship between maternal weight gain and the increasing speed of weight in different pregnant terms and macrosomia.In order to reasonably manage pregnancy and decrease the morbidity of maerosomia.Methods 106 newborns whose birth weights were equal to or greater than 4000 g were specified as macrosomia,while 106 newborn with birth weights lying in 2500-3999 g were under the control group.A case-control study was conducted to compare the corresponding factors such as maternal BMI.weight before pregnancy and the change of weight during pregnancy respectively.Results Indicated by both simple and multiple unconditional logistic regression analysis,the cause of fetal macrosomia Was mainly associated with the factors including the maternal weight before pregnancy(OR=2.204,95％CI:1.377-3.529),matemal weight gain in 12-pregnant weeks(kgper week)(OR=1.961,95％CI:1.204-3.194),maternal weight gain in 20-gestation weeks(kg perweek)(OR=1.811,95％CI:1.078-3.041),maternal weight gain in 30-pregnant weeks(kg per week)(OR=1.858,95％CJ:1.095-3.153)and virile newborn(OR=2.630,95％CJ:1.420.4.850.When in 30-pregnant weeks.the pregnant women with 0.5-1.0 kg weight gain per week had 1.13 fold risks comparing to those whose weight gains were lexq than 0.5 kg per week.Conclusion Maternal weight before pregnancy,weight gain during pregnancy and fetal sex appeared a closer relation to macrosomia.It is necessary to monitor the change of maternal weight during different pregnancy periods,especially for the 30th-pregnant weeks.

Objective To examine the prevalence of chikungunya virus in brain tissue samples from rat?like animals in Xiamen, Shenzhen and Guangzhou, and to explore whether the rat?like animals are potential sources of human chikungunya fever infections and the host of the virus. Methods Rat?like animals were trapped in residential areas, city parks, hospitals, markets and schools in Xiamen, Shenzhen and Guangzhou (Yuexiu and Baiyun districts) between January 2013 and June 2016. Brain tissue samples of the trapped animals were collected under sterile. Chikungunya virus was detected by using reverse transcription polymerase chain reaction (RT?PCR). Results Totally 1092 rat?like animals were trapped, which belonged to 7 species, 3 genera, 2 families, 2 orders. Rattus norvegicus was the dominant species in the indoor environment, Rattus losea was dominant in wild environment, and 1092 brain tissue samples were collected. No detectable chikungunya virus was found in the brain tissue samples by RT?PCR. Conclusion There is a low possibility that rat?like animals act infectious sources of human chikungunya fever infections and the host of the virus.

To explore the related risk factors and outcome in pregnancy- induced hypertension patients with Ⅲ phase of retinopathy.
●METHODS: A total of 105 pregnancy - induced hypertension patients with Ⅲ phase of retinopathy in our hospital from Januany 2012 to December 2013 were enrolled. Clinical date of them were collected to analyze.
●RESULTS: The occurrence of pregnancy - induced hypertension patients with Ⅲ phase of retinopathy were positively correlated with the course of the disease, blood pressure, proteinuria, and it was higher occurred in cold winter and spring, timely termination of pregnancy and appropriate hormone therapy can promote the recovery of vision, and improve outcomes of pregnancy.
●CONCLUSlON: The occurrence of pregnancy - induced hypertension patients with Ⅲ phase of retinopathy associated with season and disease severity. Timely treatment can restore normal vision, improve maternal and neonatal prognosis. Routine examination of fundus examination should be used as the pregnancy induced hypertension syndrome.

Objective To explore the effect of different proportions of mixed blood exchange transfusion on blood circulation in neonates with hemolytic disease.Methods Thirty-one newborn infants with hemolytic disease were treated by peripheral arteriovenous synchronization of exchange transfusion with different proportions mixed blood.AB type plasma was mixed with O type red blood cell(RBC) washing.The proportion for the treatment group was 1:1(the O type RBCs 2 U:the AB type plasma 200 mL),by exchange transfusion of haplotypes,in accordance with 80?mL/kg;the proportion for control group was 2:1(the O type RBC 4 U:the AB type plasma 200 mL),by exchange transfusion of double in accordance with 150-180 mL/kg.The indicators were detected,such as the exchange rate of neonatal serum bilirubin,RBC,hemoglobin(Hb),hematocrit(HCT),and the exchange transfusion quantity and days of hospitalization before and after the exchange transfusion were analyzed.Results The exchange rate of serum bilirubin of treatment group and control group was (44.92?3.99)% and (45.69?5.06)%,respectively,there was no significant difference between 2 groups(P=0.639),there was no significant difference of hospitalization days[(8.13?1.13) d vs(8.19?0.91) d]between 2 groups(P=0.884).After exchange transfusion in treatment group,the average level of the RBC,Hb and HCT were increased(P

Objective To construct a specific small interfering double-stranded RNA(siRNA) expression vector of Caspase-12 and to evaluate inhibitory effect of this siRNA on caspase-12 mRNA activity.Methods Three groups of siRNA targeting different gene sites of caspase-12 were designed and synthesized chemically.Mouse hepatoma cell line,Hepa1-6,was transfected with the siRNA by 24 h.Reverse transcription-polymerase chain reaction(RT-PCR)was performed to analyze the inhibi- tion of caspase-12.Then the most effective siRNA was selected and the two template sequences for the siRNA were inserted into pRNAT-H1.1Neo expression vector.The recombinant plasmid, referred to as pRNAT-casp12,was verified by PCR analysis and sequencing.The expression of caspase-12 at mRNA and protein level,after transfection with pRNAT-casp12 by 48 h and 72 h respectively,were analyzed by using real-time PCR and Western blotting.Results The chemically synthesized siRNA*1 and siRNA*3 could inhibit mouse hepatoma cell caspase-12 mRNA by 59.9% and 39.6%(P