Objective To design an instrument that can provide a series of pulse to stimulate cell by means of external electric field, the structure and function of organism can retrieve. Methods AVR MCU, produced by America ATMEL Co., is used as the core of the system. The program has been developed to adjust three pulse's parameters, including amplitude, frequency and pulse duration. The cooperation between DAC and OP completes the transformation from monopole pulse to bipolar pulse. The booster PB50 amplifies the current and voltage of the output. Results The Stimulator can provide bipolar pulse, amplitude: up to ?40V, frequency: 0.01Hz -10Hz, pulse duration: 0.4ms -24ms. Conclusion Cooperating with special electrode board, the instrument can provide effective electric field for cell simulating. At present the instrument has been used in the research of the endothelial progenitor cell.

Objectives To provide prenatal diagnosis and genetic counseling for four athigh-risk pregnant women with a suspected family or personal history of fragile X syndrome (FXS) by genetic screening of fragile X mental retardation (FMR1) gene.Methods This study was conducted on four pregnant women (No.l to 4) who received outpatient treatment in Peking University First Hospital from August 2014 to June 2016.Genomic DNA was extracted from peripheral blood samples of the pregnant women and six of their family members,four of which were suspected or confirmed FXS and the other two were FMR1 gene carriers.Amplide X kits were used to detect CGG repeat size in FMR1 gene.Two amniocytes and one chorionic villi samples were collected from three pregnant women to extract DNAs for FMR1 gene and karyotyping analyses.Results There were patients diagnosed with FXS in all the families by detecting CGG repeat numbers in FMR1 gene.The pregnant woman No.1 was a permutation carrier;No.2 carried normal FMR1 alleles while her brother had a mutation with over 20 CGG repeats in FMRI gene at chromosome X.No.3 and 4 were full mutation carriers with over 200 CGG repeats in FMR1 gene.After genetic counseling,No.3 decided to terminate the pregnancy due to abnormal fetal karyotype (47,XY,+21) and full mutation of FMR1 alleles.No.1 and 4 continued to pregnancy as their fetuses were normal in FMR1 alleles and karyotype.No.2 continued to pregnancy as her fetus was free of FXS risk.Conclusions Prenatal diagnosis and genetic counseling should be conducted on women at highrisk for FXS to avoid birth defects.People with a family history of FXS should be tested for FMR1 gene carrier status.

ObjectiveTo explore the relationship of blood volume changes and postoperative delirium in elderly patients undergoing surgery for hip fracture.Methods One hundred and fifty elderly patients who underwent surgery for hip fracture were enrolled in the study.Delirium was diagnosed by Confusion Assessment Method(CAM).Preoperative,intraoperative and postoperative data were collected,and the correlation of postoperative delirium and blood volume changes were analyzed by logistic regression analysis.ResultsA total of 59 patients(28 males and 31 females)had delirium after surgery and the occurrence rate of postoperative delirium was 39.3％ (59/150).The average age in delirium group was significantly older than that in the control group[ (77.71 ±6.63)years old vs(73.79 ±5.42) years old,t =-3.958,P ＜0.001 ].The average hemoglobin concentration and hematocrit in delirium group were both less than that in control group whether before surgery or at 7 days after surgery( before surgery:average hemoglobin concentration:[ ( 117.80 ± 16.59)g/L vs( 123.92 ±14.61 ) g/L,t =2.378,P =0.019; hematocrit:(0.355 ± 0.154) vs(0.372 ± 0.210),t =2.291,P =0.023 ;7days after surgery:average hemoglobin concentration:(98.15 ± 11.51 ) g/L vs ( 102.33 ± 9.88 ) g/L,t =2.369,P =0.019; hematocrit:(0.296 ± 0.040 ) vs (0.306 ± 0.030),t =-3.958,P ＜ 0.001 ].There was no significant difference on gender,fracture type,surgical approach,operative time,blood loss and hemoglobin concentration at 1,3 days after surgery between the two groups( P ＞0.05 ).Logistic regression analysis showed that age ( OR 3.280 ),education ( OR 0.389 ),and hemoglobin concentration at 7 days after operration ( OR 1.097) were significantly related to the occurrence of postoperative delirium ( P ＜ 0.05 ).Conclusion Our findings suggest that the risk for postoperative delirium is the result of more than one factor.Older age,continued postoperative low hemoglobin concentration and low degree of education present high risk of delirium in patients underwent surgery for hip fractures.

ObjectiveUsing the technology of siRNA to inhibit the gene expression of no-receptor　tyrosine protein kinase Lck in T cells of asthmatic mice,and to study the therapeutic effect of Lck specific　siRNA in asthmatic mice.MethodsReceptor tyrosine protein kinase Lck specific siRNA fragments were　taken from chemosynthesis.In vivo-jetPEITM was used to transfect the siRNA into mice body through tail　vein injection.The mice were killed 48 hours later,and the levels of IL-4,IL-17 in bronchoalveolar lavage　fluid (BALF) were detected with respondent ELISA kits.The change of inflammatory histopathology in lung　was observed with H.E.staining.The expression of Lck in lung was detected with immunohistochemistry　(IHC),and the level of Lck in lung tissue homogenate was detected with Western Blot.Results Compared with asthmatic group[ (234.68 ± 11.15 ) pg/ml,( 96.76 ± 8.28 ) pg/ml],the levels of IL-4,IL-17　[ (234.68 ± 11.15)pg/ml,(96.76 ±8.28) pg/ml] in the BALF of siRNA interference group decreased,　and the inflammation in the lung relieved.IHC indicated that the expression of Lck in lung decreased and　the level of Lck in lung tissue homogenate decreased ( P ＜ 0.05 ).Conclusions Lck specific siRNA　could reduce the level of IL-4,IL-17 in the lung tissues of asthmatic mice,and relieve the inflammatory reaction in lung.

Objective To perform a prenatal diagnosis for the second fetuses from 28 pedigrees with proband of mitochondrial disease due to mitochondrial DNA (mtDNA) mutation.Methods From April 2011 to November 2015,peripheral blood samples of 28 probands and their parents,urine samples of these probands and their mothers as well as amniotic fluid samples of the second fetuses from the 28 pedigrees were collected in Peking University First Hospital.DNA sequencing was used to identify mtDNA mutations.Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was performed to verify mutation sites,calculate mutation loads,and further confirm the diagnosis after birth.Microsatellite maker analysis was also performed on five short tandem repeats located in nuclear genes to exclude maternal contamination.Statistical analysis was carried out using independent t-test.Results In the 15 pedigrees carrying A3243G mutation,13 mothers and nine fetuses carried A3243G mutation.Neither the other two mothers nor their fetuses were positive for A3243G mutation.Among the 12 pedigrees with T8993G mutation,there were eight mothers carrying T8993G mutation and all of their fetuses carried the same mutation;and the other four mothers and their fetuses were negative for T8993G mutation.T10191C mutation was only found in one proband and the second fetus of that pedigree,but not in the mother.None of the fathers had mtDNA mutation.Results of PCR-RFLP were consistent with those of DNA sequencing.Short tandem repeat analysis demonstrated that amniocyte samples were from fetuses without maternal contamination.No mtDNA mutations were found in the six newborns who were negative for mtDNA mutations in prenatal diagnosis.The mean mutation load in urine samples of the six mothers without A3243G mutation in amniocytes was significantly lower than that of the nine mothers with A3243G mutation [(10.1 ±4.8) ％ vs (28.2 ± 15.1) ％,t=2.290,P=0.043].Conclusions The lower the mtDNA mutation load in maternal urine samples,the less the possibility she bears a child with mtDNA mutation.However,prenatal diagnosis of mitochondrial disease is necessary.

Aim Toinvestigatetheantagonisticeffect of intrathecal injection of carbenoxolone (CBX ) on neuropathic pain and its underlying mechanism.Meth-ods SixtymaleSprague-Dawleyratswererandomly divided into five groups (n =12 ):group I received sham surgery then treated with saline;group Ⅱ re-ceived SNT then treated with saline;groupⅢreceived SNT then treated with 0. 05 μg CBX;group Ⅳ re-ceived SNT then treated with 0. 5 μg CBX;group Ⅴreceived SNT then treated with 5 μg CBX.Treatment was undertaken with 10 μl volume as a single intrathe-cal injection on postoperative day 10.Mechanical with-drawl thresholds were measured 1 d before operation, 1,3,5,7 and 10 d after surgery,1 h before intrathe-cal administration,and 1 ,2,4,6 h after intrathecal administration.Lumbar spinal cord was obtained 2 h after intrathecal administration to determine the expres-sions of GFAP by immunohistology and TNF-α,IL-1βby ELISA in bilateral spinal dorsal horns.Results Comparedwiththeshamgroup,thebilateralMWTin group Ⅱ ~Ⅴ was significantly decreased.Compared with the MWT 1 h before intrathecal administration on day 10,the values at 1 ,2,4,6 h after administration of group Ⅱ and Ⅲ had no marked difference.The ip-silateral MWT in groupⅣhad no significant difference at 1,2,4 h after administration,the contralateral MWT was significantly increased,whereas GFAP and TNF-α,IL-1βwas significantly decreased in the spinal cord .In group Ⅴthe bilateral MWT was significantly improved at 1 ,2,4 h after administration,whereas GFAP and TNF-α,IL-1βwere significantly decreased inthespinalcord.Conclusions IntrathecalCBXcan inhibit the development of bilateral MWT.The analge-sic effect of CBX is implemented partly via suppressing the actation of GFAP and the realease of TNF-α,IL-1βin the spinal doral horn.

Objective:To summarize the characteristics of genetic variation and prenatal diagnosis in pedigrees with X-linked adrenoleukodystrophy (X-ALD) and elucidate the value of prenatal diagnosis in preventing the birth of children with X-ALD.Methods:Twenty pedigrees, clinically diagnosed with X-ALD in Peking University First Hospital from November 2012 and March 2019, were included in this retrospective study. Genomic DNA was extracted from peripheral blood and amniotic fluid or chorionic villi samples of probands and their families for detecting variants in ATP-binding cassette subfamily D member 1 ( ABCD1) gene using polymerase chain reaction (PCR)-Sanger sequencing. Linkage analysis was also performed on five microsatellite markers near ABCD1 gene to exclude maternal contamination. Characteristics of ABCD1 gene variants and prenatal diagnosis of X-ALD pedigrees were summarized by descriptive statistics. Results:Twenty ABCD1 gene variants were identified in the 20 pedigrees. The variants in three probands that were not detected by next-generation sequencing were identified by PCR-Sanger sequencing. Among the mothers of the 20 probands, 17 carried ABCD1 variants and three did not. We performed 24 prenatal diagnoses on 20 pregnancies (24 fetuses) and identified eight fetuses with variants who were finally terminated. The 16 cases without variants were born alive. The validation results obtained after termination or delivery were consistent with those performed prenatally. Conclusions:No hotspot variants in ABCD1 gene are detected in these X-ALD patients and most variants are maternally inherited. PCR-Sanger sequencing is an effective method for detecting ABCD1 variants. Prenatal diagnosis for mothers who had a body with X-ALD could prevent another one from birth.

OBJECTIVETo investigate the mutation and expression of tumor suppressor gene-PTEN mRNA and explore their roles in tumorigenesis and progression of ovarian cancer.

METHODSMutated exon 5 of PTEN gene was examined in normal ovary (n=5), ovarian cyst (n=5), ovarian borderline tumor (n=9), epithelial ovarian cancer (n=60), and ovarian cancer cell line (n=1) by polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP). mRNA expression of PTEN gene was evaluated in corresponding tissues and cell line by reverse transcription polymerase chain reaction (RT-PCR). The mutation and mRNA expression of PTEN gene were compared with clinicopathological features of ovarian cancer.

RESULTSMutated exon 5 of PTEN gene was detected only in 5 (7.1%) cases of epithelial ovarian cancer. mRNA expression level of PTEN gene in ovarian borderline tumor or ovarian cancer was lower than that in normal ovary or ovarian cyst (P < 0.05). The level of PTEN gene mRNA expression was negatively correlated with clinicopathological staging of ovarian cancer, whereas positively correlated with histological differentiation (P < 0.05). mRNA expression level of PTEN gene in ovarian endometrioid cancer was significantly lower than that in ovarian serous or mucinous cancer (P < 0.05).

CONCLUSIONSMutation of PTEN gene occurs in ovarian cancer. Down-regulated expression of PTEN is probably an important molecular event in tumorigenesis of ovarian cancer. Abnormal expression of PTEN gene is involved in progression of ovarian cancer. Reduced expression of PTEN gene is closely associated with tumorigenesis and pathobiological behaviors of ovarian endometrioid cancer.

Adolescent ; Adult ; Aged ; Carcinoma, Endometrioid ; enzymology ; genetics ; Down-Regulation ; Exons ; Female ; Gene Expression Regulation, Neoplastic ; Genes, Tumor Suppressor ; Humans ; Middle Aged ; Mutation ; Ovarian Neoplasms ; enzymology ; genetics ; PTEN Phosphohydrolase ; Phosphoric Monoester Hydrolases ; biosynthesis ; genetics ; Polymerase Chain Reaction ; Polymorphism, Single-Stranded Conformational ; RNA, Messenger ; genetics ; Tumor Suppressor Proteins ; biosynthesis ; genetics

OBJECTIVETo investigate the expression of PTEN and Caspase-3 in malignant lymphoma of the stomach and explore their role in progression of primary gastric malignant lymphoma.

METHODSFormalin-fixed paraffin embedded tissues from 56 cases of primary gastric malignant lymphoma and their adjacent non-tumor mucosa were evaluated for PTEN and Caspase-3 protein expression by streptavidin-biotin-complex (SABC) immunohistochemistry. Their expression was compared with clinical tumor parameters with the relationship between PTEN and Caspase-3 expression concerned as well.

RESULTSThe positive rate of PTEN expression in primary gastric lymphomas (50.0%, 28/56) was significantly lower than that in adjacent non-tumor gastric mucosa (96.4%, 27/28) (P < 0.05). Meanwhile, 43 of 56 (76.8%) gastric lymphomas indicated Caspase-3 expression, less than that in adjacent non-tumor mucosa (93.5%, 29/31) (P < 0.05). The expression of PTEN was negatively correlated with invasion and lymph node metastasis of gastric lymphoma (P < 0.05), while the Caspase-3 expression was negatively associated with the latter one (P < 0.05). Additionally, the PTEN expression was positively correlated with Caspase-3 expression in the primary gastric malignant lymphoma (P < 0.05).

CONCLUSIONSThe down-regulated expression of PTEN and Caspase-3 played an important role in progression of primary malignant gastric lymphoma. PTEN, as a molecular marker of pathobiological behaviors of tumor, contributes to tumor progression by increasing cell mobility and angiogenesis, as well as decreasing cell adhesion and apoptosis.

Adult ; Aged ; Biomarkers, Tumor ; metabolism ; Caspase 3 ; Caspases ; metabolism ; Down-Regulation ; Female ; Gastric Mucosa ; enzymology ; Humans ; Lymphatic Metastasis ; Lymphoma ; enzymology ; pathology ; Male ; Middle Aged ; Neoplasm Invasiveness ; PTEN Phosphohydrolase ; Phosphoric Monoester Hydrolases ; metabolism ; Stomach Neoplasms ; enzymology ; pathology ; Tumor Suppressor Proteins ; metabolism

The aim of this study is to study the damage effects of chronic hypoxia on medulla oblongata and to explore whether the damage is associated with oxidative stress and cell apoptosis. Adult male SD rats were randomly divided into two groups: control group and chronic hypoxia group. Medulla oblongata was obtained for the following methods of analyses. Nissl's staining was used to examine the Niss bodies of neurons in medullary respiratory related nuclei, biochemistry methods were utilized to examine oxidant stress damage induced by chronic hypoxia on medulla oblongata through measuring malondialdehyde (MDA) content and superoxide dismutase (SOD) activity, and RT-PCR technique was used to study the influence of apoptosis induced by chronic hypoxia on medulla oblongata through analyzing the levels of Bax mRNA and Bcl-2 mRNA. The results showed the optical densities of Nissl's staining in pre-BötC, NA, NTS, FN, and 12N were significantly decreased in chronic hypoxia group in comparison with that in control group (P < 0.05). In chronic hypoxia group, MDA level was significantly higher than that in the control group (P < 0.05), whereas SOD level had no significant difference between the two groups (P > 0.05). Bax mRNA expression had no obvious change and Bcl-2 mRNA expression significantly decreased in chronic hypoxia group in comparison with that in control group (P < 0.05). The results suggest that chronic hypoxia could bring about serious damage to medullary respiratory centers through aggravating oxidative stress and increasing cell apoptosis.
Animals ; Apoptosis ; Chronic Disease ; Hypoxia ; physiopathology ; Male ; Medulla Oblongata ; metabolism ; pathology ; physiopathology ; Oxidative Stress ; Rats ; Rats, Sprague-Dawley ; Respiratory Center ; metabolism ; pathology ; physiopathology ; Superoxide Dismutase ; metabolism