Abstract:Objective:To evaluate the impact of preimplantation genetic diagnosis procedures on the heat stress response in the offspring using mouse model. Methods:We use the mouse models,which divided into PGD group and Normal group. At 5 weeks of age,evaluate the stress system of PGD offspring under 45℃ heat stress. Results:Under heat stress,the mean death time is 59.31 min and 79.63 min in PGD and Normal group, respectively(P < 0.001). There is no significant difference between the two groups of mice under the stress,such as serum hormones,structure and neuronal activity of locus coeruleus and hypothalamus. Similarly,there is no significant difference of adrenal structure between two groups,and the level of adrenaline,noradrenaline and corticosterone is the same trend. Conclusion:The different response ability between two groups under heat stress is not related to the function of locus coeruleus,hypothalamus and adrenal. It may be related to the further function of their secretioned hormones.

Abstract:Objective:To study the effect and mechanism of miR-106a on the development of Alzheimer’s disease (AD) induced by amyloid beta protein (Aβ) in primary cultured hippocampus neurons and PC12 cells. Methods:The primary cultured hippocampus neurons and PC12 cells were treated with Aβ. The expression of miR-106a,STAT3 and JAK2 mRNA were detected by RT-PCR,while the expression of STAT3,p-STAT3 and JAK2 protein were evaluated by Western blot. Results:The expression of miR-106a was lower in Aβ-treated primary cultured hippocampus neurons and PC12 cells than that of controls (P < 0.05). In addition,the expression of STAT3 mRNA and protein was higher in Aβ-treated cells than that of controls (P < 0.05),while the expression of JAK2 showed no significant difference whether Aβ-treated was used or not (P > 0.05). Conclusion:This study suggested that miR-106a might contribute to the pathogenesis of AD by negatively regulating the expression of STAT3.

Abstract:Objective:To obtain the anthrax protective antigen in E. coli,prepare its monoclonal antibodies,and analyze the function. Methods:The gene coding for anthrax protective antigen PA10 was synthesized and cloned into pUC57 plasmids. After being identified by enzyme analysis,the target gene was inserted into pColdⅡ plasmid. By sequencing confirmation,the plasmid pCold II-PA10 was transformed into E. coli BL 21(DE3) and induced by IPTG and low temperature for expression,and identified by SDS-PAGE. The recombinant protein was purified by affinity chromatography and identified by Western blotting. After the mice BALB/c were immunized by the recombinant protein of PA10 as antigen,the monoclonal antibodies were prepared. Results:The recombinant protein of PA10 was used to immunize mouse,and obtain 4 strains of specific monoclonal antibodies(respectively named as 2G8, 5A8, 7B3,9C9). The protection of anthrax toxin test in vitro,proved that one of four monoclonal antibody strains(7B3) has a comparatively large activity to neutralize anthrax toxin with protection rate up to 96%. Conclusion:This research successfully constructed the anthrax Protective antigen PA10 in E. coli and prepared its monoclonal antibodies,which laid the foundation for further development of chimeric neutralizing antibodies used to treat anthrax.

Abstract:Objective:To observe the effect of the solube egg antigen (SEA) on B cell function and the related signal transduction. Methods:In vitro, mouse spleen CD19+ B cells were stimulated by SEA or LPS for 72 hours. Then the expression of CD80, CD86 and CD40 and cell cycle were analyzed by flow cytometry, and the dilution of CFSE was assayed by flow cytometry. At the same time, the IL-10, IL-6, IFN-γ and TGF-β level in cultured supernatants were detected by ELISA. PD98059 (ERK inhibitor), SP600125 (JNK inhibitor), SB203580 (p38MAPK inhibitor) and PDTC (NF-κB inhibitor) were used to study the signal pathways for the induced secretion of IL-10, IL-6 in B cells. Results:The expression of CD80, CD86 and CD40 was up-regulated in B cells stimulated by SEA or LPS. And the ratio of B cells entered S/G2-phase was increased after stimulated by SEA or LPS. SEA or LPS promote B cell proliferation. The IL-10 and IL-6 secretion increased in mouse spleen B cells culture supernatants stimulated by SEA or LPS (P < 0.01). Furthermore, PD98059, SP600125, SB203580 and PDTC inhibited the cytokine secretion in B cells stimulated by SEA or LPS. Conclusion:SEA could up-regulate and promote co-stimulatory molecules expression on B cells and the B cells proliferation. At the same time, SEA could induce the IL-10 and IL-6 secretion of B cells through NF-κB, ERK, JNK and p38MAPK signal transduction pathway.

Abstract:Objective:To elucidate the function of melatonin on hypoxia -induced epithelial- mesenchymal transition (EMT) of SGC-7901 gastric cancer cells and to reveal the mechanisms underlying. Methods:After hypoxia for 96h,expression of epithelial and mesenchymal makers of SGC-7901 gastric cancer cells was detected using Western blot and immunofluorescence was employed to investigate distribution of proteins to see if EMT acured. Then we treated SGC-7901 gastric cancer cells by 1 mmol/L concentration of melatonin under hypoxia environment to see if EMT was inhibited. Results:Hypoxia induced morphological change of SGC-7901 gastric cancer cells,as well as decreased E-cadherin and increased vimnetin and α-SMA. The reactive oxygen species ( ROS) and exprssion of hypoxia-inducible factor-1α(HIF-1α) both abolished by treatment of melatonin,so as EMT. Conclusion:We found that melatonin can abolish hypoxia-induced EMT of SGC-7901 gastric cancer cells,which can be used for clinical application.

Abstract:Objective:To investigate functional characteristics of coding region of solute carrier family 26,member 4(SLC26A4)gene and to further study the correlation between its mutations and occurrence of non-syndromic hearing loss(NSHL). Methods:Bioinformatic methods were applied to analyze the phylogeny of Pendrin,protein coded by SLC26A4 among 37 species. Conserved regions were identified by a sliding window method. Prediction of secondary structure and homologous modeling were conducted by online tools. Results:SLC26A4 gene was highly conserved among mammals,especially among primate. Twelve conserved regions were identified by using a sliding window method. Prediction of secondary structure suggested that coiled-coils may exist in N- and C-terminals. Homogenous modeling indicated that the intervening sequence(IVS) within the STAS domain varies among different species. Conclusion:Mutations of SLC26A4 play a vital role in the occurrence of NSHL and missense mutations in conserved regions are more likely to cause structural and functional changes,thus generating corresponding phenotypes. IVS within the STAS domain of C-terminal may have special functions in human.

Abstract:Objective:To explore the mRNA expression of EphA5 receptor in hippocampus of neonatal rat with hypoxic-ischemic encephalopathy (HIE) as well as the relationship with apoptosis of neurons. Methods:Seven-day-old SD rats were randomly divided into HIE group,HIE+ EphA5-Fc group and control group . The mRNA expression of EphA5 receptor and the apoptosis of neurons in hippocampus of neonatal rats in 3 groups were determined by qRT-PCR and TUNEL staining. Results:①The mRNA expression of EphA5 receptor in HIE group increased and peaked at 12h,which was still higher than that in control group at 48 h (P < 0.05). Compared with HIE group,the mRNA expression of EphA5 receptor decreased in HIE+ EphA5-Fc group (P < 0.05),which recovered to that in control group (P > 0.05). ②In HIE group,apoptosis of hippocampus neurons increased after 6 h,and peaked at 24 h. Apoptosis of hippocampus neurons in HIE+ EphA5-Fc group was remarkably lower than that in HIE group (P < 0.05). Conclusion:The up-regulation of EphA5 receptor and apoptosis of hippocampus neurons could be suppressed by inhibitor of EphA5 receptor,which suggests the activity of EphA5 receptor may be one of the mechanisms of brain damage by hypoxic-ischemic.

Abstract:Objective:To study and compare the gene expresse of Oct4 and Nanog in human pancreatic cancer stem cell. Methods: FCM was used to isolate pancreatic cancer stem cells of CD44+CD24+ and CD44+CD24+ESA+ phenotypes by a FACS Aria II in human cell line Panc-1. Immunofluorescence was used to detect the expression of Oct4 and Nanog gene in Panc-1 cell and two kinds of cancer stem cell-like cells. Results:Oct4 and Nanog genes have a higher expression in CD44+CD24+ESA+ cells and CD44+CD24+ cells than Panc-1 cells. Conclusion:Oct4 and Nanog gene perform a higher expression in pancreatic cancer stem cells than in Panc-1 cells.

Abstract:Objective:To investigate the correlation between neuroendocrine(NE)differentiation and clinico-pathological features,prognostic factors of papillary thyroid carcinoma(PTC). Methods:The tissues of 103 PTC cases were amaylzed by HE staining,immunohisto-chemistry,immunogold labelling,biomarkers expression of neuroendocrine differentiation. Results:Thyroglobulin was positively expressed in PTC,while Calcitonin was negative. The larger tumor size of the cases with PTC accompanied with a higher NSE expression(P < 0.05). The higher degree of differentiation accompanied with a lower rate of CgA expression. The cases with nuclear inclusions showed obviously increased expression of CgA(P < 0.05). The degree of PTC cell differentiation and tumor size were associated with NE differentiation under TEM(P < 0.05). The CgA immunogold particles were scattered at the internal edge of NE particles within the cytoplasm of PTC cells. Conclusion:NE differentiation could occur in PTC,and NE differentiated cells could be derived from follicular epithelial cells instead of C cells. NE differentiation is correlated with tumor size,morphological change of nuclei and degree of differentiation in PTC. NE differentiation has no significant correlation with the prognosis of PTC.

Abstract:Objective:To observe the curative effect of noninvasive positive pressure ventilation (NIPPV) in the treatment of severe COPD-induced type II respiratory failure combined with acute left heart dysfunction. Methods:Eighty-six COPD patients with acute left heart dysfunction were randomly divided into control group (n = 43) and treatment group (n = 43). The control group was treated with routine treatment,and the treatment group was treated with bi-level positive airway pressure (BiPAP) ventilation through a nasal or face mask in addition to routine treatment. For the treatment group,continuous BiPAP ventilation was given in the first 2 days. Since the third day,the patients were given 8 to 20 hours BiPAP ventilation. BiPAP ventilation was gradually weaned when patient＇s condition stabilized. Changes of heart rate,blood gas indexes,left ventricular ejection fraction (LVEF),NT-proBNP,and clinical symptoms (including dyspnea,conscious state,pulmonary rales) in the two groups before and after the treatment were compared. Results:The heart rate,pH,PaO2,PaCO2 in the treatment group after BiPAP ventilation for 2 h,24 h,48 h,and 7 d were significantly improved compared to those of the control group (all P < 0.05). The LVEF values of 7 days after BiPAP ventilation in the treatment group were significantly increased than that of the control group. Moreover,the NT-proBNP levels of the treatment group were significantly decreased compared to the control group after 48 hours and 7 days treatment. Conclusion:BiPAP ventilation had a good therapeutic effect for patients with severe COPD-induced type II respiratory failure combined with acute left heart dysfunction.

Abstract:Objective:To evaluate the prognosis of senile critical patients over 75 years old by continuous monitoring blood lactic acid and D-dimer. Methods:One hundred and twenty-six critical patients over 75 years old were included in this research,and divided into two groups:improved surviving group and poor prognostic death group. Blood lactic acid and D-dimer were continuously monitored for the first week at admission into ICU. Results:Of 55 cases in the surviving group,APACHE II score was 18.8 ± 4.5 at admission in ICU,meanwhile lactic acid was (1.7 ± 1.3) mmol/L,and D-dimer was (2.03 ± 1.55) mg/L. Of 71 cases in the death group,APACHEⅡ score at admission was 25.3 ± 6.8,lactic acid was (2.4 ± 1.6) mmol/L,and D-dimer was (2.36 ± 1.07) mg/L. At admission,only APACHEⅡ score in the death group was significantly higher than that in the surviving group. After a week stay in ICU,lactic acid was (1.2 ± 0.6) and (5.6 ± 4.4) mmol/L,D-dimer was (2.05 ± 0.81) mg/L and (3.19 ± 1.68) mg/L in the surviving and the death group, respectively. Both indicators were significantly higher in the death group. Conclusion:Continuously monitored lactic acid and D-dimer were important indicators to evaluate the clinical prognosis of senile patients. The prognosis would usually be better if both lactic acid and D-dimer keep decreasing during the first week at admission in ICU.

Abstract:Objective:To investigate the expression level of miR-518d in the placenta and maternal serum of gestational diabetes mellitus and normal pregnant women. Methods:The expression of miR-518d in placenta and serum was measured by Taqman RT-PCR in 20 women with GDM and 20 women with normal term pregnancy. Results:Expression of miR-518d in placenta was significantly up-regulated in GDM group(4.35 ± 0.88)compared to control group(1.04 ± 0.73)(P < 0.01). Expression of miR-518d in serum was also up-regulated in GDM group(2.43 ± 1.07)compared to control group(0.96 ± 0.88)(P < 0.05). We also showed that PPARa was a direct target of miR-518d with a specific binding site at the seed sequence. And expression of miR-518d in the placenta was significantly positive correlated with the expression of serum E2(R2=0.759,r = 0.871). Conclusion:miR-518d is related with the mechanism of GDM and we suggest that upregulation of miR-518d may be associated with the pathogenesis of GDM via an effect on the regulation of PPARα expression.

Abstract:Objective:To compare the influences of two different diagnostic criteria of Gestational Diabetes Mellitus (GDM) on detection rates and pregnancy outcomes. Methods:More than ten thousand cases of pregnant women undergoing glucose screening and delivery in Nanjing Maternal and Child Health Care Hospital were enroll in this study and divided into two groups. One group was diagnosed according to the criteria in 7th edition of “Obstetrics and Gynecology”,and the other group was diagnosed according to the criteria of International Diabetes and Pregnancy Study Group (International Association of Diabetes and Pregnancy Study Groups,the IADPSG). We compared the GDM detection rates,mode of delivery,common complications of pregnancy and neonatal complications incidence between the two groups. Results:The GDM detection rate of IADPSG group was significantly higher than that of the 7th edition of “Obstetrics and Gynecology” group (P < 0.05). The cesarean section rate of IADPSG diagnostic criteria was significantly lower than the those of 7th edition of “Obstetrics and Gynecology” group (P < 0.01). The incidences of polyhydramnios and premature rupture of membranes were significantly lower than those of the 7th edition of “Obstetrics and Gynecology” group (P < 0.05). There were no significant differences of the incidences of hypertensive disorders on pregnancy,diabetes ketoacidosis,fetal distress and postpartum hemorrhage between two groups(P > 0.05). The incidences of fetal macrosomia,neonatal hypoglycemia and neonatal respiratory distress syndrome polyhydramnios were significantly lower than those of the 7th edition of “Obstetrics and Gynecology” group (P < 0.05). There were no significant differences of the incidences of premature birth,low birth weight infants,neonatal asphyxia,neonatal jaundice,neonatal hyperbilirubinemia,neonatal developmental malformations between two groups (P > 0 05). Conclusion:The diagnosis of GDM according to the IADPSG criteria can effectively elevate the detection rate of GDM,dramatically reduce the incidences of polyhydramnios,premature rupture of membranes and fetal macrosomia,thereby reducing the rate of cesarean section,and significantly reduce neonatal hypoglycemia and newborn respiratory distress and improving maternal and neonatal outcomes.

Abstract:Objective:To investigate the relationship between serum 25-Hydroxy Vitamin D and lipid profiles among middle-aged and elderly population. Methods:Individuals of 724 were enrolled in this study. Fasting blood samples were collected and serum 25-Hydroxy Vitamin D and biochemical indices were determined. Subjects were divided into three groups (T1-T2-T3) according to tertiles of 25-Hydroxy Vitamin D levels. ANOVA was used to detect the difference among the three groups. Multiple linear regression was conducted to investigate relationships between 25- Hydroxy Vitamin D levels and lipid profiles. Results:There was significant difference among three groups for TG,but not for TC,HDL and LDL. 25-Hydroxy Vitamin D was associated with TG in total population(regression coefficient -0.013,P < 0.001). After adjustment for age,BMI,fasting blood glucose,HbA1c,glutamic transaminase,and γ-glutamyl transpeptidase,the association was still significant in all subjects and females. Conclusion:Middle-aged and elderly individuals with lower serum 25-Hydroxy Vitamin D had higher TG level. 25-Hydroxy Vitamin D is associated with TG negatively,and particularly in females. However there was no significant association between 25-Hydroxy Vitamin D and TC,HDL or LDL.

Abstract:Objective:To study the relation shipbetween lung function and bone mineral density(BMD),serum Vitamin D in elderly patients with chronic obstructive pulmonary disease(COPD). Methods:Pulmonary function,BMD of lumbar spine and femoral neck,and serum Vitamin D were determined in 104 COPD(61 male,43 postmenopausal female) patients and 100 cases age and sex matched non-COPD controls. The association of lung function and BMD with serum vitamin D was analyzed as well. Results:Vitamin D insufficient and osteopenia were present in elderly men.The prevalence of Vitamin D deficiency and osteoporosis was higher in COPD group than that in non-COPD controls (37.5% vs 16.0% and 38.5% vs 20.0%)(P < 0.05). The increase of COPD severity led to the decrease of average values of BMD of femoral neck (P < 0.05),as well as Vitamin D (P < 0.05). Correlation analysis showed that BMD and Vitamin D were correlated to FEV1(P < 0.05). Conclusion:Prevalence of Vitamin D deficiency is high in COPD patients,increases with the severity of COPD,and is closely associated with osteoporosis prevalence. BMD and Vitamin D were correlated to pulmonary function.

Abstract:Objective:To compare the variation of titanium implant artifacts in different sequences and optimize scanning parameters for high-quality images. Methods:Titanium water phantom was designed for routine sequences and those with optimized parameters scanning;the variation of artifacts was observed with altering field of view,echo train length,bandwidth in sagT2-FSE sequence. Results:Artifacts were changed under different sequences. The reduction of field of view,the decrease of echo train length and bandwidth reduced the artifacts. Conclusion:Change of the scan sequence or scan parameters could reduce artifacts caused by metal implants,improve image quality,and benefit clinical diagnosis.

Abstract:Objective:To explore the value of diffusion tensor imaging(DTI)in radiologically evaluating cerebral small vessel disease(SVD). Methods:Forty-three cases of SVD were imaged with conventional MRI and DTI. Fifty-one lesions in 43 cases were recruited in our study. Average diffusion coefficient(DCavg)and fractional anisotropy(FA)were measured symmetrically and statistically analyzed,which were at level of the lesion and the posterior limb of internal capsule and the cerebral peduncle of the affected and contra-lateral sides. The structure images of cortical spinal tracts(CST)were obtained by diffusion tensor tractography(DTT). Results: There presented three conditions of CST on DTT:CST was showed to run naturally and have a complete structure form in 41/51 lesions,and to be pressured and shifted in 9/51 ones and interrupted in 1 lesion. Both DCavg and FA values of the affected side were different significantly from those of the contra-side(P < 0.05),and the DCavg value is negatively associated with FA value(P < 0.05). The FA values between chronic lacunar infarction and chronic ischemic focus were significantly differentiated(P < 0.05). Conclusion:DCavg and FA values of lesions in SVD changed regularly,and DTT could have a great contribution to estimate the extent of damage to CST in SVD.

Abstract:Objective:To explore the clinical values of 131I whole-body scan (131I-WBS) and 131I single photon emission computed tomography/computed tomography (131I-SPECT/CT) on the management of patients with differentiated thyroid carcinoma (DTC) after 131I therapy,respectively. Methods:After 3~5 days 131I therapy,48 DTC patients underwent 131I-WBS and 131I-SPECT/CT. The imaging features and clinical values of 131I-WBS and 131I-SPECT/CT were comparatively analyzed. Results:The patients had totally 48 131I-WBS and 60 131I-SPECT/CT. 145 foci were observed by 131I-WBS,including 34.3% of foci localized in the thyroid bed,60.5% of malignant lesions,and 5.2% caused by nonthyroid physiologic or benign uptake or a contaminant. There were 23 additional metastases detected by 131I-SPECT/CT. Futhermore,there were 20 patients whose 131I-WBS showed 65 inconclusive lesions. Precise localization and characterization by 131I-SPECT/CT fusion images were 65(100 %) and 61(94 %) of the 65 foci,respectively. 131I-SPECT/CT fusion images were considered to be of much benefit to 18(37.5%) of 48 patients. Changes of therapeutic plans happened to 8(44.4%) of 18 patients. Conclusion:131I-WBS should be routinely performed in the management of patients with DTC after 131I therapy. If 131I-WBS images were inconclusive or inconsistent with the clinical manifestations,131I-SPECT/CT must be required to increase the diagnostic accuracy and modulate therapeutic plans.

Abstract:Objective:To evaluate the clinical application of submental artery island flap in reconstruction of hypopharynx following hypopharyngectomy. Methods:Five patients with pyriform sinus squamous cell carcinoma were enrolled. All of patients were male,ages ranged from 49 years to 70 years with a mean age of 61.2 years. Among them,4 patients underwent selective neck dissection along with the radical excision of the primary lesion,and the laryngeal function was partly reserved. The submental artery island flaps were then used for reconstruction of hypopharyngeal defects in these 4 patients. Another patient was anastomosis orifice stricture of postoperative pyriform sinus carcinoma,and the reconstruction using submental artery island flap was performed following fissure of the stricture. The size of the skin paddle ranged from 7 cm × 3 cm to 8 cm × 5 cm. Results:The surgical procedures were successful,and the submental artery island flaps were survived in all cases. Donor site incision healed by first intention. Trachea cannula and gastric tube were pulled-out 7~36 days after operation. The follow-up period ranged from 10 to 28 months. The statuses of the flaps were good with satisfactory speech and swallowing function in all the patients. Neck scar contracture was well hidden. Conclusion:The submental artery island flap is useful in hypopharyngeal reconstruction after resection of hypopharyngeal carcinoma,because of its abundant blood supply,convenient donor site,and high survival rate.

Abstract:Objective:To study the application of serratus anterior muscle flap transplantation in expander-based implant breast reconstruction and its clinical significance. Methods:Forty-seven breast cancer patients received modified radical mastectomy or mammectomy (a total of 47 breast) were enrolled. They have taken staging breast reconstructoin using soft tissue expanders (SRTX). Then the clincal results have been observed,recorded and analyzed. The serratus anterior muscle flap was used as lateral coverage in the operation. The complications such as postoperative capsular contracture,seroma,hematoma,incision dehiscence and infection have been recorded in both periodⅠ and periodⅡ operations. Results:Of all patients (47 cases),there was no complication such as hematoma,infection,or skin flap necrosis happened during the follow up after periodⅠ and periodⅡ operations. Only 2 cases (2 sides) had incision dehiscence after periodⅠ. During the follow-up period(465.0 ± 2.5) days,no capsular contracture happened. Conclusion:Using anterior serratus muscle flap in expander-based implant breast reconstruction can fix the expander and implant,reduce complications,prevent breast implant displacement and create good breast shape.

Abstract:Objective:To introduce a minimally invasive percutaneous nephroscopic necrosectomy experience for infected necrotizing pancreatitis. Methods:Two patients with serve acute pancreatitis received minimally invasive percutaneous nephroscopic necrosectomy in our hospital during 2012. Before the minimally invasive percutaneous nephroscopic necrosectomy along the drainage sinus tract,the first patient underwent one time of percutaneous catheter drainage,the second patient underwent three times of percutaneous catheter drainage and one time of open debridement. Results:Two patients received one time of percutaneous nephroscopic treatment,symptoms of infection was quickly under control. There were no operative mortality and morbidity. Conclusion:Percutaneous nephroscopic necrosectomy is a minimally invasive,safe and efficient treatment for infected pancreatic necrosis when used in selected patients.

Abstract:Objective:To investigate the association between nonsynonymous polymorphisms (rs1039808,rs12508222,rs1567047 and rs1014867) in FAT4 gene and the risk of gastric cancer. Methods:A case-control study of 783 gastric cancer cases and 855 controls was conducted in a Chinese population of Jiangsu Province using the TaqMan allelic discrimination assay on the ABI 7900 system. Results:We found that A allele of rs1039808 (Ala807Val) was significantly associated with a decreased risk of gastric cancer risk in additive model after adjusted by age,sex,smoking and drinking status (OR = 0.74,95%CI:0.62~0.88,P = 0.001). Similar association strengths were shown between all subgroups for rs1039808(P>0.05 for heterogeneity test). Conclusion:These findings indicate that the nonsynonymous variants rs1039808 (Ala807Val) of FAT4 may contribute to gastric cancer susceptibility in a Chinese population of Jiangsu Province.