Objective To investigate the role of mitogen-activated protein kinases (MAPK)/extracellular signal regulated kinase1/2 (ERK1/2) signal transduction pathway in regulating the expression of aquaporin 3 (AQP3) in human amnion epithelial cells.Methods Primary cell culture of human amnion epithelial cells deriving from amnion of term pregnancy with normal amniotic fluid volume (AFV) or isolated oligohydramnios was conducted in the Second Affiliated Hospital of Wenzhou Medical College from January to November 2011.Either group included 10 elective cesarean cases.The primarily cultured cells were treated with different concentrations (0,5,10,20 and 40 mol/L) of ERK inhibitors (U0126) for 12 h,and then the optimal concentration of U0126 which resulted in the lowest expression of phospho~ERK1/2 (p-ERK1/2) was added for different durations(0,2,6,12 and 24 h).Immunocytochemistry was used to detect the localization of AQP3 and Western blot analysis was used to examine the expression of total ERK1/2,p-ERK1/2 and AQP3 in human amnion epithelial cells.Statistical analysis was performed by t-test and one-way ANOVA.Results (1) Compared with those in normal AFV group,p-ERK1/2 and AQP3 expression in human amnion epithelium cells decreased in oligohydramnios group,respectively (p-ERK1/2:3.46 ± 0.33 and 2.46±0.25;AQP3:2.34 ± 0.18 and 1.56±0.10,t=9.243 and 13.292,P＜0.01).(2) In oligohydramnios groups,after treated with different concentrations of U0126,the expressions of total ERK1/2 did not change (F=0.365,P＞0.05).The expression of p-ERK1/2 and AQP3 in 5,10,20,40 μmol/L U0126 (p-ERK1/2:0.96±0.16,1.12±0.13,0.98±0.17 and 1.02±0.26; AQP3:1.10±0.09,1.12±0.08,1.13±0.06 and 1.11±0.06) were all significantly lower than those in 0 μmol/LU0126 group (p-ERK1/2:2.46±0.25; AQP3:1.56±0.10,P＜0.05).However,the expression of p-ERK1/2 and AQP3 showed no significant difference among 5,10,20,and 40μmol/L U0126 groups (P＞0.05).The optimal concentration of U0126 was 5 μmol/L.After treated with 5 μmol/L U0126 for 2 h,the expressions of p-ERK1/2 and AQP3 (1.27±0.29 and 1.44±0.12)were lower than those after treated for 0 h (2.55±0.12 and 2.15±0.09,P＜0.05).However,there was no significant difference among groups treated for 2,6,12 and 24 h.Therefore,the optimal treatment time was 2 h.(3) The expression of AQP3 was distributed in both cell membrane and cytoplasm in amnion epithelial cells with normal amniotic fluid volume or isolated oligohydramnios,but mainly in cytoplasm.U0126 did not change the localization of AQP3 expression.Conclusions U0126 inhibits the phosphorylation of ERK1/2 and expression of AQP3 of women with oligohydramnios,indicating that the MAPK/ERK1/2 signal transduction pathway might regulate the expression of AQP3 in human amnion epithelial cells,and therefore affect the balance of amniotic fluid volume.

Objective To investigate the expression of aquaporins-3 (AQP3) in amniotic epithelial cells regulated by cyclic adenosine monophosphate-protein kinase A (cAMP-PKA) signal pathway and to explore the mechanisms of its expression.Methods The amniotic epithelial cells were collected from 30 patients who underwent elective caesarean sections at term with normal amniotic fluid volume and primarily cultured.The cultured cells were treated with (1) forskolin groups: different concentration (0,2.5,5,50 or 100 μmol/L) of forskolin treated cells for 2 hours,and the optimal concentration of forskolin treated cells with different time (0,1,2,10 or 20 hours) ; (2)SP-cAMP groups: different concentration (0,2.5,5,50 or 100 μmol/L) of SP-cAMP treated cells for 2 hours,and the optimal concentration of SP-cAMP treated cells with different time (0,1,2,10 or 20 hours); (3)H-89 groups: different concentration (0,5,10,50 or 100 μmol/L) of H-89 treated cells for 2 hours,and the optimal concentration of H-89 treated cells with different time (0,1,2,10 or 20 hours).The level of intracellular cAMP and activity of PKA were detected by using ELISA,and immunohistochemistry was used to detect the localization of AQP3,the protein expression of total cAMP-response element binding protein (CREB) and phospho-CREB (p-CREB) and AQP3 were assessed by western blot analysis.Cell proliferation was assessed by cell counting kit-8 (CCK-8)assay.Results (1) The brown staining of AQP3 was detected in both cell membrane and cytoplasm in each group.(2) There was no significant change of the cell proliferation rate among groups with different concentration of forskolin,SP-cAMP and H-89 treatment (P ＞ 0.05).(3) After different concentration of forskolin treated 2 hours,the expression of total CREB had no significant difference among them(P ＞ 0.05).While the expression of cAMP level,PKA activity,p-CREB and AQP3 protein were significantly changed,which were higher in 2.5 μmol/L,5 μmol/L,50 μmol/L forskolin group when compared with 0 μmol/L (P ＜ 0.05).Their expressions in 5 μmol/L forskolin group were higher than that in 2.5 μmol/L and 50 μmol/L (P ＜ 0.05).The optimal forskolin concentration was 5 μmol/L.(4) After different concentration of SP-cAMP treated 2 hours,the expression of total CREB and cAMP level had no significant difference among them (P ＞ 0.05),while the expression of PKA activity,p-CREB and AQP3 protein were significantly changed,which were higher in 5 μμmol/L,50 μmol/L SP-cAMP group when compared with 0 μmol/L (P ＜ 0.05).Their expressions in 50 μmol/L SP-cAMP group were higher than that in 5 μmol/L (P ＜0.05).The optimal SP-cAMP concentration was 50 μmol/L (5) After different concentration of H-89 treated 2 hours,the expression of total CREB and cAMP level had no significant difference among them (P ＞ 0.05),while the expression of PKA activity,p-CREB and AQP3 protein were significantly changed,which were lower in 10 μmol/L,50 μmol/L and 100 μmol/L H-89 group when compared with 0 μmol/L (P ＜ 0.05).Their expressions in 10 μmol/L H-89 group were lower than that in 50 μmol/L,100 μmol/L (P ＜ 0.05).The optimal H-89 concentration was 10 μmol/L.(6) p-CREB and AQP3 protein expression were significantly lower in 5 μmol/L forskolin combined 10 μmol/L H-89 incubating 2 hours group when compared with 5 μmol/L forskolin,but higher than that in 10 μmol/L H-89 treated group (P ＜ 0.05).Total CREB was no significant difference among the three groups (P ＞ 0.05).Conclusion cAMP-PKA signal transduction pathway may regulate AQP3 protein expression in human amniotic epithelial cells.

Objective To observe the effect of acupuncture at nape points plus nerve electrical stimulation on deglutition function and cerebral blood flow in pseudobulbar palsy after cerebral stroke. Method The eligible subjects were randomized into a control group of 35 cases and a treatment group of 33 cases. The control group was intervened by conventional Western medication, while the treatment group was by acupuncture at nape points plus nerve electrical stimulation in addition to the Western medication, once a day, 4 weeks as a treatment course. Result The total effective rates were respectively 93.9%and 80.0%in the two groups, and the treatment group was significantly superior to the control group; after the intervention, the high whole blood viscosity, low whole blood viscosity, hematocrit, and plasma viscosity all dropped markedly in the treatment group, and were significantly different from that in the control group (P<0.05). The Transcranial Doppler (TCD) showed that the blood flow mean velocity (Vm), systolic velocity (Vs), and pulsatility index (PI) of the left vertebral artery, right vertebral artery, and basilar artery in the treatment group were significantly improved after the intervention (P<0.05), and the Vm and Vs of the left vertebral artery, right vertebral artery, and basilar artery in the control group were significantly improved after the intervention (P<0.05), while PI in the control group didn’t show significant improvement (P>0.05). After the intervention, there were significant differences in comparing the Vm, Vs, and PI of the left vertebral artery, right vertebral artery, and basilar artery between the two groups (P<0.05). Conclusion Acupuncture at nape points plus nerve electrical stimulation is effective in improving the deglutition function in pseudobulbar palsy after cerebral stroke, and its action mechanism is plausibly through improving the cerebral blood flow and promoting the recovery of brain function.

This paper develops the field medical information management system,which realizes the informationiztion and team service management for field medical team.system to field medical team. It also provides interface program with No.1 Military Medical Project,which realizes the linking of medical treatment information about patient in field medical team and rear military hospital.

The visual acuity testa were performed in 8219 children(16 438 eyes)under 5 years of age with sound and light toys,electric torch and a retinoscope.The general ocular examination and mydriatic optometry were also performed for the ametropia.In 8219 children 61 were found to have dominant strabismus,including 59 cases of concomitant strabismus and 2 paralytic strabismus.AbnormaJ refraction was detected in 404 children(739 eye)accounting for 4.92%,including 68 cases of anisometropia (0.83%).In 16 438 eyes 621(3.78%)were moderate or hyper-presbyopia;53(0.32%)were myopia;65 (0.40%)were mixed astigmatism.The vision acuity test described above is a simple and convenient method suitable for children under 5 who are not able to read the conventional test chart.

Objective To detect fibrillin-1 gene (FBN1) mutations in patients with Marfan syndrome (MFS) by denaturing high-pedormance liquid chromatography (DHPLC) and DNA sequencing. Methods Genomic DNA was extracted from whole blood sample of 22 MFS patients. All 65 exens of FBN1 were amplified by polymerase chain reaction(PCR) respectively. Mutations were screened by DHPLC followed by DNA sequencing of the PCR products which showed different DHPLC profiles from the normals. Results Ten mutations of the FBN1 were found in 9 MFS patients. The mutations comprised four missense[5015G > C(C1672S),5309G > A(C1770Y),7241G > A(A2414G) and 7769G > A(C2590Y)], four nonsense [3295G > T ( E1099X ), 430"/insTCGT (G1441X), 4621C > T ( R1541X ) and 8080C > T (A2694X)], and two splice site mutations (IVS29 + 4A > T and IVSSO + 1G > A). Conclusion It is suggested that DHPLC coupled with DNA sequencing is an efficient method for the detection of FBN1 gene mutations, and it may be useful in diagnosis of MFS.

Objective To investigate outcome and prognosis of isolated mild fetal ventriculomegaly (IMV) of fetus in uterus. Methods From Jan. 2006 to Dec. 2009, 18 200 singleton pregnancy women from 20 weeks gestation underwent prenatal ultrasonography examination in Department of Obstetrics and Gynecology, Second Affiliated Hospital of Wenzhou Medical College. One hundred and forty-eight women with IMV (transverse diameter of the atrium of the lateral ventricle measuring between 10 and 15 mm with no other abnormalities) were studied prospectively, which were divided into two groups: 99 women with transverse diameter of the lateral ventricle of 10 - 11 mm in group A and 49 women with transverse diameter lateral ventricle of 12 - 15 mm in group B. The changes of ventriculomegaly and the associated intracranial and extracranial anomalies were observed regularly every 2 or 4 weeks until delivery. The development of neurological system was also followed up. Results ( 1 ) The overall incidence of IMV was 0. 08% (148/18 200). The rate of bilateral ventriculomegaly were 20% (20/99) in group A and 51% (25/49) in group B, which reached statistical difference (P＜ 0. 05). (2) Prognosis of fetus: 139 cases with 2 or more ultrasonographic examinations, IMV resolved throughout pregnancy in 41. 7% (58/139) ,regressed in 7. 9% (11/139) ,remained stable in 36. 7% (51/139)and progressed in 13. 7% ( 19/139). Five cases in group A and 11 cases in group B present progress, which reached significantly difference (P ＜ 0. 05). (3) One hundred and eleven cases infant were followed up for 5-12 months,the rate of psycho-motor developmental delay was 5. 4% (6/111). The rate of neuro-developmental delay in progressed group (3/15) was higher than 2. 5% ( 1/40) in resolved group, 0 (0/8) in regressed group and 4. 2% (2/48) in remained stable group, which reached significantly difference (P＜0. 05). Conclusions About 85% of cases of IMV resolved, regressed or remained stable in utero would exhibited good prognosis. IMV with a transverse atrial size ≥ 12 mm or progression in utero was usually associated with a poor prognosis, which should be observed carefully.

The aging process of human colonic epithelium involves a slow decline in physiological vigor and an increasing susceptibility to age-related diseases, especially, colon cancer, but the molecular mechanisms of the aging and susceptibility of aged colonic epithelium to carcinogenesis is still unclear. Identification of aging related proteins in colonic epithelium will help to reveal the molecular mechanisms of colonic epithelial aging and age-related colonic diseases. Therefore, the total proteins of human normal colonic epithelial tissues from 10 young and 10 old men were separated by two-dimensional gel electrophoresis(2-DE), respectively. PDQuest software was applied to analyze 2-DE images, the differentially expressed protein spots of colonic epithelium between young and old groups were identified by matrix-assisted laser desorption/ionization time of flight mass spectrometry(MALDI-TOF-MS), and the expression levels of partial identified proteins were determined by real-time quantitative PCR and immunohistochemistry. Well-resolved, reproducible 2-DE maps of human colonic epithelial tissues from young and old men were established, 17 aging related proteins were identified by MALDI-TOF-MS, and the differential expression levels of partial identified proteins were confirmed by real-time quantitative PCR and immunohistochemistry. The results indicate that injury of mitochondrial function and decline of antioxidant capability are important reasons for the aging of human colonic epithelium, and four differential proteins (guanine nucleotide-binding protein beta subunit-like protein, stress-70 protein, 40 S ribosomal protein SA and chloride intracellular channel protein1) may be involved in susceptibility of aged colonic epithelium to carcinogenesis.

Objective Hepatitis A immunization strategies were carried out in 2001 in Tianjin. We wanted to evaluate the effectiveness of the strategies related to hepatitis A control programs and to provide the basis for further modification of the strategies. Methods Descriptive epidemiology study was used to analyze the hepatitis A epidemic situation in 2000-2011 in Tianjin and to evaluate the disease reporting system. Hepatitis A vaccine coverage of target population and serum epidemiological study were carried out in 1999,2005 and 2010 to check on the hepatitis A antibody levels so as to evaluate the immuno-barrier condition in the normal population. Cox-Stuart test was used to analyze the epidemic trend of hepatitis A and other intestinal infectious diseases in Tianjin. Results The incidence rate of hepatitis A decreased from 2.89/100 000 in 2000 to 0.12/100 000 in 2011,and the percentage of hepatitis A in all types of viral hepatitis decreased from 8.02%in 2000 to 0.48% in 2011 in Tianjin. The positive rates of hepatitis A antibody also increased in the residents. Conclusion The hepatitis A vaccination program was successful in the programs on prevention and control of hepatitis A in Tianjin,China.

Objective To evaluate the effect of deferoxamine on ventilator-associated lung injury in rats.Methods Twenty-four healthy male Sprague-Dawley rats,aged 6-8 weeks,weighing 250-300 g,were divided into 3 groups (n =8 each) using a random number table method:control group (group C),ventilator-associated lung injury group (group VALI),and ventilator-associated lung injury plus deferoxamine group (VALI+DFO group).Normal saline 2 ml was intraperitoneally injected in C and VALI groups,and deferoxamine 200 mg/kg (dissolved in 2 ml normal saline) was intraperitoneally injected in group VALI+DFO.The animals were connected to a small animal ventilator 15 min later and mechanically ventilated in volume-controlled mode,with tidal volume 40 ml/kg,respiratory rate 40-60 breaths/min,inspiratory/expiratory ratio 1 ∶ 1,and inspired oxygen fraction ratio 1.0.The rats were sacrificed after the end of mechanical ventilation,and the left lung tissues were removed for examination of the pathological changes (with a light microscope) which were scored and for determination of wet/dry weight ratio (W/D ratio).The right lung was lavaged,and lavage fluid was collected to prepare macrophage suspension,and the alveolar macrophage and mitochondrial reactive oxygen species (ROS) levels were determined using flow cytometry.Results Compared with group C,the pathological score,W/D ratio of lung tissues,and alveolar macrophage and mitochondrial ROS levels were significantly increased in group VALI,and the pathological score was significantly increased in group VALI (P＜0.05).Compared with group VALI,the pathological score,W/D ratio of lung tissues,and alveolar macrophage and mitochondrial ROS levels were significantly decreased in group VALI and DFO (P＜0.05).Conclusion Deferoxamine can reduce ventilator-associated lung injury,and the mechanism may be related to inhibiting oxidative stress in rats.