Objective: To obtain chimeric antigen receptor macrophages ( CAR-M) targeting HER2 stably transfected． Methods : CAR lentivirus vector targeting HER2 was constructed and infected with human monocytic leukemia cell line (THP-1) ．CAR THP-1 cells with green fluorescent labeling were selected by sorting flow cytometry and continued to be cultured in vitro．The CAR THP-1 cells targeting HER2 were co-cultured with the endometrial cancer cell line Ishikawa with negative and positive HER2 expression，and their targeted phagocytosis of CAR-M to HER2 positive tumor cells was detected by imaging flow cytometry ，and the targeted phagocytosis efficiency of CAR-M to HER2 positive tumor cells was detected by flow cytometry. Results : CAR lentivirus infection with THP- 1 cells was less efficient ; After co-culture with cancer cells，flow cytometry and imaging flow cytometry showed that CAR THP-1 cells had enhanced phagocytosis of HER2 positive Ishikawa cells compared with the empty body group (P＜0. 01) ． Conclusion In this experiment，CAR THP-1 cell line targeting HER2 was established by constructing CAR lentivirus vector and transfecting THP-1 cells ，and it was proved that CAR THP-1 could phagocytize HER2 positive Ishikawa cells through specific targeting.

A method of high performance liquid chromatography coupled with time-of-flight mass spectrometry was used to detect the endogenous metabolites changes in plasma of normal rats, rats of dampness stagnancy due to spleen deficiency and Astragalus flavone component intervention rats. Metabolism map of rat plasma was obtained and the mechanism of Astragalus flavonoids on dampness stagnancy due to spleen deficiency was studied. Rat model with dampness stagnancy due to spleen deficiency was established by high fat and low protein diet plus load swimming. Liquid chromatography tandem mass spectrometry was used for the analysis of rat plasma sample, and 0.05％ formic acid water with 0.05％ formic acid acetonitrile as the mobile phase was applied in gradient elution with Halo C18 chromatographic column. In this study, partial least squares discriminant analysis and variance analysis were used to screen the potential biomarkers, it was found that the metabolic profile of the Astragalus flavonoids was different from that of the model group, which was close to that of the normal group. A total of 11 potential biomarkers were identified, including glycerol phospholipids, sphingolipids, amino acids, and so on. The metabolic pathways of biomarkers including three tricarboxylic acid cycle, glycerol phospholipid metabolism, fatty acid metabolism, amino acid metabolism and so on, which mainly related to energy metabolism and fat metabolism in the body. Related indexes of rats with syndrome of spleen deficiency of water and dampness were significantly callback after Astragalus flavone intervention, including macro indicators such as body weight, independent activities and micro indicators such as metabolic markers, blood lipids and others. The result showed that Astragalus flavonoids played the role of strengthening the spleen and draining the water mainly through regulating the energy metabolism, fat metabolism and so on.

An App for Android intelligent mobile phone to recite prescriptions in rhyme and learn infused pre-scriptions of traditional Chinese medicine was developed in order to meet the requirement of students majoring in traditional Chinese medicine and lovers of traditional Chinese medicine, which can show the assistant and guide of prescriptions in rhyme by different colors. An interface linked with each corresponding traditional Chinese medicine was added into the App to show its picture, properties, effect and application for the interaction between database server and mobile phone users and the aim to recite prescriptions of traditional Chinese medicine and learn tradi-tional Chinese medicine.

Objective To evaluate the residual risk (i.e.failure risk in detecting aneuploidies abnormalities except for chromosome 13,18,21,X and Y) of cytogenetic abnormalities using interphase fluorescence in situ hybridization (FISH) for the second-trimester amniocytes.Methods The results of interphase FISH and conventional karyotyping of 2 837 consecutive amniotic fluid specimens were analyzed retrospectively.Probes for chromosomes 13,18,21,X and Y were used.The detection rate and residual risk for interphase FISH were calculated for the following three major clinical indications for prenatal diagnosis (advanced maternal age,abnormal maternal serum screening indicating an increased risk for trisomy 18 or trisomy 21,and ultrasound abnormalities).Results Consecutive interphase FISH and karyotyping of second-trimester amniocytes for prenatal diagnosis were performed from January 1,2010 to July 31,2013.Among the 2 837 cases,85 (3.0％) cases with abnormal karyotypes were found,including 73 cases of aneuploidies involving chromosome 13,18,21,X and Y,which were considered detectable by interphase FISH; 12 cases of chromosomal anomalies,other than aneuploidies of chromosome 13,18,21,X and Y,were diagnosed after karyotyping and were not detected by interphase FISH,including six cases of balanced rearrangements,five cases of imbalanced rearrangements,and one case of pseudomosaic of trisomy 20.Of these 12 chromosomal anomalies,three cases of imbalanced rearrangements involving chromosome 21 showed positive FISH results,and the other nine cases showed negative FISH results among which four case of hereditary balanced rearrangemerts and two cases of novel balanced rearrangements.The total detection rate for interphase FISH was 89.4％ (76/85),the misdiagnosis rate of chromosome abnormalities was 14.1％(12/85),and the residual risk was 0.43％ (12/2 761) following interphase FISH of the second-trimester amniocytes.Conclusions Interphase FISH is a useful adjunct to conventional karyotyping,but should not be regarded as a replacement for karyotyping as too many structural chromosomal abnormalities will be missed.Providing patients with a detection rate and residual risk during counselling may help them understand the advantages and limitations of interphase FISH in their prenatal diagnostic evaluation.

Objective To investigate the effects of microRNA-29a (miR-29a) on lipopolysaccharide (LPS)-induced apoptosis in human monocytes THP-1 cells in order to understand the molecular mechanisms.Methods Human monocytes THP-1 cell line were exposed to LPS after transfected with miR-29a inhibitors (100 nmol/L) or just transfected with miR-29a mimic (100 nmol/L) by lipofectamine RNAiMAX.Flow cytometry (FCM) was used to detect the cell apoptosis.Real-time RT-PCR was employed to measure expressive levels of the gene Bcl-2 and Mcl-1.The luciferase assay was performed in HEK293T cells,which were co-transfected with plasmid DNA and miRNA by using Lipofectamine 2000.Statistical analysis carried out by using SPSS 13.0 software for One-way ANOVA and Student' s t test.Results Transfection with miR-29a mimics for 48 h increased apoptosis rate and significantly reduced the expressions of Bcl-2 and Mcl-1 in THP-1 cells in comparsion with the control group.The apoptosis rate also raised in THP-1 cell stimulated by LPS for 24 h followed by LPS stimulation for 24 h,the apoptosis rate was decreased in comparison with the LPS group.In addition,our luciferase assay data showed that HEK293T cells cotransfected with miR-29a mimics and Bcl-2 3 ' UTR-Wt or Mcl-1 3' UTR-Wt plasmid significantly reduced the luciferase activity compared with the control group.Conclusions The miR-29a may regulate apoptosis by targeting the genes Bcl-2 and Mcl-1,and miR-29a may play a pivotal role in the process of apoptosis in immune cells.

Objective To investigate the relationship between abnormalities of maternal serum markers in Down syndrome screening in second trimester and adverse pregnancy outcome.Methods Totally,1935 pregnant women were screened for Down syndrome with maternal serum tri marker with time-resolved fluorescence assay,including alpha fetoprotein (AFP),free β-human chorionic gonadotropin (β-hCG) and unconjugated estriol (uE3),between 15 and 20+6 gestational weeks at Peking Union Hospital from January 1,2009 to January 31,2011,and were followed up till delivery.The relationship between incidence of adverse pregnancy outcomes and women with normal or abnormal levels of serum markers in Down syndrome screening was investigated.T-test or Chi-square test were applied for statistical comparison.Results (1) Among the 1935 pregnant women,normal levels of serum markers were found in 1255(normal group) and 680 were abnormal(abnormal group),in which 577 with only one abnormal serum marker,89 with two and 14 with three abnormal serum markers.According to the serum marker level,the 577 women with one abnormal serum marker were further divided into five groups,including high AFP group (n=17),low AFP group (n=114),high β-hCG group (n=242),low β-hCG group (n=139) and low uE3 group (n=65).The birth weight of infants in lower AFP group and the gestational age at delivery in low β-hCG group were greater than those in normal group [(61.3±9.1) kg vs (59.5±8.3) kg,(272.6±11.8) d vs (274.4±10.1) d,t=2.21 and 1.99,both P＜0.01].(2) The incidence of adverse pregnancy outcome in normal group was 42.8％(537/1255),while comparing with the abnormal group(43.7％,297/680),no statistical significance was shown (RR =1.02,P =0.71).While comparing with the normal group,the incidences of placenta previa [25.5％ (32/1255) vs 2/17,RR=4.61,P＜0.05] and abnormal placental morphology were higher in high AFP group [4.1％ (51/1255) vs 5/17,RR=7.24,P＜ 0.05],the incidence of gestational diabetes mellitus (GDM) was higher in low AFP group [8.1％ (101/1255) vs 14.4％(16/114),RR=1.74,P＜0.05],the incidence of placenta and membrane retention was higher in high β-hCG group [3.5％ (44/1255) vs 6.2％(15/242),RR=1.77,P＜0.05],the incidence of pre-eclampsia was higher in low β-hCG group [1.7％ (21/1255) vs 6.5％ (9/ 139),RR=3.87,P＜0.05].(3) There were 89 women with two abnormal serum markers.Comparing with the normal group,the incidences of small for gestional age (SGA) infants,oligohydramnios,abruptio placenta were higher in women with low AFP but high β-hCG [SGA infants:6.9％(2/29) vs 1.8％(22/1255),RR=3.94; oligohydramnios:20.7％(6/29) vs 6.4％(80/ 1255),RR=3.24; both P＜0.05],the incidences of oligohydramnios was higher in women with both low AFP and low uE3[3/14 vs 6.4％(80/1255),RR=3.36,P＜0.05],the incidence of premature birth and GDM were higher in women with both low β-hCG and low uE3 [premature birth:2/6 vs 4.3％(54/1255),RR=7.75; GDM:3/6 vs 8.0％ (101/1255),RR=6.21; both P＜0.05].(4) There were 14 women with three abnormal serum markers.The relationship between adverse outcome and abnormal serum markers did not show any statistical significance.Conclusions The abnormality of serum markers of Down syndrome screening is closely related to adverse pregnancy outcomes,and women with abnormal serum markers should be carefully monitored during pregnancy.

Objective To calculate the incidence of chromosomal abnormalities at second trimester in women who were 35 or older at their expected date of birth.Methods The amniocentesis and karyotyping results in Peking Union Medical College Hospital from January 1st,2001 to June 30th,2011 were retrospectively analyzed.The only indication for amniocentesis in these group of woman was advanced maternal age.A total of 6584 cases Were included in this study and were divided into two groups according to maternal age,ie.35-39 and ≥40 year old group.The incidences of fetal 47,+ 21,47,+ 18 and sex aneuploidies were calculated and compared between two groups by Chi-square test.Results Altogether,121 cases were diagnosed to be abnormal chromosome,and the overall incidence was 18.38‰ (121/6584).The abnormal karyotypes included 111 cases of aneuploidies (mosaicism included) and 10 cases of structural abnormalities.The aneuploidies included 59 cases of 47,+21 (8.96‰,59/6584),25 cases of 47,+18 (3.80‰,25/6584),2 cases of 47,+13 (0.30‰,2/6584) and 25 cases of sex aneuploidies (3.80‰,25/6584).Fetal 47,+21 was the most frequent chromosomal abnormality,accounting for 53.15％ (59/111) of all aneuploidies.The incidence of fetal 47,+21 was significantly higher in ≥40 year-old group than that of 35-39 year old group[13.99‰(16/1144) vs 7.90‰(43/5440),x2=3.937,P=0.047].There were no statistical differences of the incidences of fetal 47,+ 18 and sex aneuploidies between the two groups.Conclusions The main fetal chromosomal abnormalities in women aged 35 and older are the aneuploidies of chromosome 21,18,13 and sex chromosomes.The incidence of fetal 47,+21 is significantly increased in the women aged 40 years and older.So prenatal screening should be provided first to women at 35-39 years of age and amniocentesis should be the first choice of prenatal diagnosis for women over 40 years old.

Objective To discuss the application value of transesophageal echocardiography (TEE) in occluding procedure with cardiopulmonary bypass (CPB) for children muscular ventricular septal defects (mVSD).Methods Forty-nine mVSD patients for occluding procedure with CPB were performed guided and monitored by TEE,9 cases were simple mVSD,40 were mVSD combined with other cardiac malformations.The location,size,number,scope and the surrounding anatomy of mVSD were carefully observed before going into cardiac arrest,preparing the operator to choose appropriate occluder devices.The procedure was evaluated after the recovery of heart beat using TEE.Results In all of the 49 patients underwent occluding procedure,44 cases were implanted into one occluder,5 implanted into two occluders.The results of TEE after operation showed that the positions of all occluders were not shifted,2 had trace residual shunt,there were no complications worsening,such as mitral and aortic regurgitation,etc.One patient abandoned treatment for serious infection,the other patients' occluders were all in normal position followed up at 6 months to 2 years after operation,there were no tiny residual shunt and valves regurgitation except 1 patient.Conclusions The technique that TEE guiding and monitoring in occluding procedure with cardiopulmonary bypass for children muscular ventricular septal defects can shorten the time of cardiopulmonary bypass,significantly reducing complications and risks of the surgery,improving the successful rate.

Objective To investigate the the relationship of a high risk serum screen for Down syndrome in second trimester and adverse pregnancy outcomes,and to evaluate the predictive value for adverse pregnancy outcomes.Methods The tri-marker second trimester maternal serum screening for Down syndrome (alpha-fetoprotein,free beta-hCG and unconjugated estriol)was performed on the pregnant women at Peking Union Medical Hospital from January 2009 to January 2011.The cutoff valvue was 1/270.Pregnancy outcomes were followed up.The general condition and pregnancy complications of the pregnant women with high risk (high-risk group) were compared to that of the pregnant women with low risk (low-risk group); and with 35 years old as a demarcation,the incidences of adverse pregnancy outcomes were calculated in the two groups.Results ( 1 ) A total of 1935 cases were collected.And 1784 cases were with low risk,and 151 cases were with high risk.The difference of weight and gestational age betweem the two groups was not statistically significant ( P ＞ 0.05 ) ; the difference of age between the two groups was statistically significant ( P ＜ 0.01 ).(2) Pregnancy complications were found in 791 cases.In high-risk group,the incidences of gestational diaetes mellitus (GDM,13.9％),neonatal asphyxia (4.0％ ) and small for gestational age infant ( SGA,4.6％ ) were higher than that in low-risk group ( 8.4％,1.0％,1.6％ ),the difference was statistically significant ( P ＜ 0.05 ).The incidences of gestational hypertension disease,premature labor,oligohydrammios,placenta previa,placenta abruption,fetal macrosomia in the two groups was not statistically different (P ＞0.05).(3) In 1705 cases aged less than 35 years,129 cases (7.6％) were GDM,43 cases ( 2.5％ ) were gestational hypertension disease,61 cases ( 3.9％ ) were premature labor; in 230 cases aged 35 years or more,41 cases (17.8％ ) were GDM,12 cases (5.2％) were gestational hypertension disease,15 cases (6.5％ ) were premature labor,and the difference between the two groups was statistically significant ( P ＜ 0.05 ).In ＜ 35 years old group,the incidences of GDM,neonatal asphyxia and SGA (12.3％,4.4％,5.3％ ) were higher in the high-risk group than that (7.2％,0.9％,1.6％ ) in the low-risk group,and the difference was statistically significant ( P ＜ 0,05 ).In ≥35 years old group,the incidences of GDM,neonatal asphyxia and SGA ( 18.9％,2.7％,2.7％ ) were slightly higher in the high-risk group than that (17.6％,1.6％,1.6％ ) in the low-risk group,the difference between the two groups was not statistically significant (P ＞ 0.05 ).Conclusions The present study revealed apparertt increase in the adverse pregnancy outcomes in women with a high risk of Down syndrome screening test.Advanced age is the most important risk factor for a high risk of Down syndrome screening test and adverse pregnancy outcomes.More attention should be attached to the patients whose age were ＜35 years old and with a high risk of Down syndrome screening test.

Background: Anal fissure is one of the most common anal-rectum diseases, and approximately 10 percent patients with chronic anal fissure ultimately receive surgery. Relieving postoperative pain and protecting functions of the sphincter are central issues for coloproctologists. Objective: To evaluate the efficacy and safety of anoplasty in the treatment of chronic anal fissures. Design, setting, participants and interventions: In this prospective, multicenter, randomized controlled trial, 120 adult patients with chronic anal fissure were referred from Department of Coloproctology of Yueyang Hospital of Integrated Traditional Chinese Medicine Affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai Ninth People's Hospital Affiliated to Shanghai Jiao Tong University School of Medicine and Shanghai Municipal Hospital of Traditional Chinese Medicine. The patients were enrolled from January 2009 to April 2010 and randomly divided into study (mucosa advancement flap anoplasty, abbreviated as anoplasty) group and control (fissurectomy) group. The two groups were assessed separately, and the main outcome measures were observed for 2 weeks, with a short-term follow-up for 6 weeks. Main outcome measures: Degree of pain, haemorrhage and anal canal pressure were observed and recorded preoperatively, and on the third day, the fourteenth day and the sixth week postoperatively. The wound healing time was also recorded. Surgical complications of the two groups were recorded and compared on the third day and the sixth week postoperatively. The curative effects associated with the surgery were analyzed on the fourteenth day and the sixth week after surgery and the therapeutic results were evaluated. Results: Three patients were dropped out due to the early discharge from hospital and losing connection (1 in study group and 2 in control group). Overall the surgery showed that the anoplasty group had better results than the fissurectomy group in the curative effect on the sixth week after operation (P<0.05). Time of wound healing in the anoplasty group was (17.22±4.41) d and was better than (21.24±7.44) d of the fissurectomy group (P<0.05). Concerning the relief of wound pain, the anoplasty group achieved better results than the fissurectomy group at the third day, the fourteenth day and the sixth week after operation (P<0.05). Anoplasty reduced bleeding and had better efficacy than the fissurectomy at the third day and the fourteenth day after operation (P<0.05), however, there was no statistical difference at the sixth week after operation (P>0.05). There were no significant differences in relieving the anal canal pressure (P>0.05) and the surgical complications (dysuria, edema of anal margin, fever, infection, anal incontinence and anal deformation) between the two groups (P>0.05). None of the patients suffered postoperative complications by the sixth week after operation. Furthermore, there was no recurrence in either of the two groups at six weeks after operation. Conclusion: The results indicate that anoplasty for chronic anal fissures has advantages such as better therapeutic effects, less postoperative pain, a shorter healing time and no incidence of anal incontinence.