Objective To investigate the influence of intensive antiplatelet scheme assisted with gamma-immunoglobulin on clinical efficacy, the relieve time of clinical symptoms and the levels of inflammatory factors of children with Kawasaki disease.Methods80 children with Kawasaki disease were chosen in the period form July 2014 to July 2016 in Yiwu hospital of Zhejiang province and randomly divided into two groups including the control group (40 cases) with bigeminy scheme including aspirin and dipyridamole and the observation group (40 cases) with trigeminy scheme including aspirin, dipyridamole and clopidogrel on the basis of gamma-globulin;and the clinical efficacy for short-term, the relieve time of clinical symptoms and signs, the the levels of WBC, PLT, PCT, CRP, IL-6, TNF-α, HMGB1 and MIF before and after treatment, the incidence of coronary artery injury and the incidence of adverse drug reactions of both groups were compared.ResultsThe total clinical effects of observation group were significantly higher than control group(P<0.05).The relieve time of symptoms and signs of observation group were significantly shorter than control group(P<0.05).The levels of WBC and PLT after treatment of observation groups were significantly lower than control group and before treatment(P<0.05).The levels of PCT, CRP, IL-6, TNF-α, HMGB1 and MIF after treatment of observation groups were significantly lower than control group and before treatment(P<0.05).The incidence of coronary artery injury of observation groups were significantly lower than control group(P<0.05).There was no significant difference in the adverse drug effects incidence between 2 groups.ConclusionIntensive antiplatelet scheme assisted with immunoglobulin in the treatment of children with Kawasaki disease can efficiently relieve the respiratory symptoms and signs, control the levels of inflammatory reaction, inhibit the platelet aggregation, prevent the occurrence of coronary artery injury and not cause the serious adverse drug reactions.

Objective:To determine the relationship between trimester-specific gestational weight gain and pregnancy outcome among women with twin pregnancies.Methods:A retrospective cohort study was conducted among Chinese women with twin pregnancies delivered from January 2013 to October 2015 in the International Peace Maternity & Child Health Hospital of China Welfare Institute.Gestational weight gain was compared between women with different progestational BMI.After the adjustment for age and progestational BMI,binary Logistic regression analysis was conducted to explore the relationship between gestational weight gain rate and gestational complications,premature birth,premature rupture of membrane and neonatal birth weight.Results:①A total of 472 subjects met the inclusion criteria.Rate of gestational weight gain were(0.41 ±0.15),(0.64 ±0.30) and (0.49 ±0.15) kg/week in Early,Mid-and Late-,and Whole of pregnancy respectively.②With regards to trimester-specific gestational weight gain,statistically significant associations were found between rate of weight gain and gestational hypertension and preterm birth (P ＜ 0.05).Those with faster gestational weight gain had higher rate of premature rupture of fetal membrane(P ＜ 0.05).And those with lower gestational weight gain had lower neonatal birth weight(P＜0.05).③After adjusting maternal age,progestational BMI and gestational age at delivery,the sum of twins' birth weight increased 25.21g,30.89g and 21.46g respectively when the maternal body weight increased 1kg during the whole pregnancy,in Early and Mid-pregnancy (P ＜ 0.001),and during in Late-pregnancy(P =0.001).Conclusions:Gestational weight gain among women with twin pregnancy are closely related with adverse pregnancy outcomes.The gestational weight gain rate in different gestation are of some predictive value for pregnancy outcomes.More research with long follow-up are needed to determine proper gestational weight gain for twin pregnancy suitable for Chinese women.

Objective To assess the variations in different thyroid stimulating hormone(TSH) and free thyroxine (FT4) detection kits for evaluating thyroid function during pregnancy and to establish the corresponding normal reference ranges.Methods This study was based at the International Peace Maternity and Child Health Hospital affiliated to Shanghai Jiaotong University School of Medicine.A total of 200 pregnant women who visited the hospital between June,2011 and September,2012 were recruited in this study according to the National Academy of Clinical Biochemistry (NACB) criteria.Blood samples were sequentially collected from the women at the first (T1,9-12 weeks),second (T2,16-24 weeks) and third (T3,32-36 weeks) trimesters to determine the serum TSH and FT4 levels using four different detection kits (Siemens-C,Siemens-Ⅰ,Abott and Roche).A linear trend test was used to analyze serum TSH and FT4 levels with four different kits.A percentile range of P2.5 to P97.5 was used to establish the normal trimester-dependent reference ranges of TSH and FT4 levels for different detection kits.The Bootstrap method was used to compare the differences in the four reference ranges.Results Similar dynamic changes in TSH and FT4 levels during pregnancy were detected among the different kits (F=0.950,P=0.595; F=11.640,P=0.081,respectively).Among the four reference ranges of TSH,the Roche kit showed the most remarkable fluctuation during pregnancy,while Roche kit in the first trimester and Siemens C kit in the second and third trimesters showed larger fluctuations in reference ranges of FT4.More importantly,the reference ranges of TSH and FT4 showed significant variations among the four different kits in each trimester (TSH:T1:F=2 945.390,P ＜ 0.01; T2:F=2 826.260,P ＜ 0.01; T3:F=1 698.360,P ＜ 0.01.FT4:Tl:F=1 145.440,P ＜ 0.01; T2:F=2 260.240,P ＜ 0.01; T3:F=1 439.920,P ＜ 0.01).Conclusions TSH and FT4 measurement using four different commercial kits showed similar trimester-dependent dynamic changes.However,it is necessary to establish trimester-dependent and detection kit dependent normal reference ranges of TSH and FT4 for thyroid function evaluation for pregnant women.

Objective To investigate the mechanisms of nitric oxide (NO) in decreasing intestinal mesenteric arterial hypocontractility in rats with hepatic cirrhosis and portal hypertension,and to analyze the interaction of NO and RhoA/ROCK pathway.Methods The levels of NO in the peripheral blood and mesenteric artery of normal rats (normal control group,5 rats),rats with portal hypertension (experimental control group,6 rats)and rats with portal hypertension treated by L-NAME (L-NAME group,6 rats) were detected.Mesenteric arteriole contractility to norepinephrine in the 3 groups was determined using a vessel perfusion system.The expressions of proteins of NO-cGMP-PKG pathway and RhoA/ROCK pathway in the 3 groups were detected by Western blot.All data were analyzed using the analysis of variance or LSD-t test.The changes of mesenteric arteriole contractility to norepinephrine was expressed in dose-response curve,and was analyzed using the nonlinear regression method,and the EC50 value was calculated.Results (1) The pressures of portal veins of the normal control group,experimental control group and L-NAME group were (6.2 ± 0.9)mm Hg (1 mm Hg =0.133 kPa),(13.9 ± 1.7)mm Hg and (16.6 ± 1.3) mm Hg,respectively,with a significant difference among the 3 groups (F =94.4,P ＜ 0.05).(2) The levels of NO in the normal control group,experimental control group and L-NAME group were (43 ± 5) μmol/L,(82 ± 16) μmol/L and (45 ± 9) μmol/L,respectively,with a significant difference among the 3 groups (F =24.77,P ＜ 0.05).The level of NO of the L-NAME group was significantly lower than that of the experimental control group (P ＜ 0.05).(3) The levels of NO in the mesenteric artery of the normal control group,experimental control group and L-NAME group were (236 ±41) μmol/g,(407 ± 82) μmol/g and (216 ± 42) μmol/g,respectively,with a significant difference among the 3 groups (F =20.29,P ＜ 0.05).The NO level of the L-NAME group was significantly lower than that of the experimental control group (P ＜ 0.05).(4) Compared with the experimental control group,the dose-response curve of mesenteric arterioles to norepinephrine shifted to the left,while it did not reach the level of the normal control group.The levels of EC50 of the normal control group,experimental control group and the L-NAME group were 6.458 × 10-7 mol,9.546 × 10-7 mol and 7.494 × 10-7 mol,respectively.There was a significant difference in the EC50 level between the L-NAME group and the other two group (t =2.726,3.112,P ＜ 0.05).(5) Compared with the normal control group,the protein expression levels of eNOS and p-VASP of mesenteric artery of the experimental control group were significantly increased (P ＜ 0.05),while they were decreased in the L-NAME group (P ＜ 0.05).The protein expression levels of eNOS and p-VASP of mesenteric artery of the L-NAME group were significantly higher than those of the normal control group (P ＜0.05).There were no obvious changes of protein expression levels of PKG-1,ROCK-1 and p-moesin in the 3 groups (P ＞ 0.05).(6) The activity of ROCK-1 was significantly increased with norepinephrine stimulation in the normal control group and the L-NAME group,while no obvious changes were detected in the experimental control group.Conclusions The NO expression is upregulated in mesenteric arteries in rats with hepatic cirrhosis and portal hypertension.Such changes induce ROCK activation via influencing the expression of vasoconstrictors.L-NAME can reduce the NO levels in the mesenteric arteries,which may improve RhoA/ROCK signal pathway transduction.This can help vasoconstrictors induce ROCK activation without affecting the protein expression of ROCK.

Objective To evaluate the clinical efficacy and safety of endoscopic submucosal dissection (ESD) guided with endoscopic ultrasonography (EUS) for rectal neuroendocrine neoplasms(NENs).Methods A retrospective analysis was performed on 58 patients with rectal ENEs who underwent ESD from January 2011 to December 2015 in JiangSu Province Hospital.Manifestations of EUS, clinicopathological characteristics, proliferation activity grade, complete resection rate, complications and follow-up results of lesion were studied.Results Those treated by ESD included 58 patients with 64 lesions of rectal NENs.EUS results showed that 3 lesions originated from mucosa, 3 from muscularis mucosa and 58 from submucosa.A total of 34 lesions located within 5 cm from anus, 26 in 6-10 cm from anus and 4 more than 10 cm from anus.All 64 lesions were successfully treated by ESD.The mean maximum diameter of the lesions was 0.8 cm(0.2-3.5 cm), and the mean procedure time was 31 min(10-60 min).The complete resection rate was 93.8% (60/64).There were 4 patients with positive basal surgical margin, and two of them underwent additional surgery and two others were treated with argon plasma coagulation after rejecting surgery and ESD.Histological examination determined that 59 lesions were pathologic grade 1(G1) and 5 were pathologic grade 2(G2).Delayed bleeding occurred in 4 cases after ESD,which was managed by medicine in 1 case and endoscopic treatment in 3 cases.No perforation occurred after ESD.During a mean follow-up period of 22.9 months(3-48 months), no lymph node metastasis or distant metastasis was observed.Conclusion EUS is able to distinguish the origin of rectal NENs and aid determining the range and depth of ESD.ESD appears to be a safe, feasible and effective procedure for providing accurate histopathologica1 evaluations as well as curative treatments for rectal NENs limited to submucosa.

Fetal growth restriction is a serious high-risk pregnancy, which threats the fetal health in gestation and also increases the long-term likeliness of several diseases. Multiple factors result in this disease. Placenta is an important organ to maintain fetal growth, metabolism, maternal and fetal physiologic balance. The correlation between placental factors and FGR plays an important guiding role on seeking for the causes, pathogenesis and treatment of the disease. In this review, the authors summarized the findings in current studies of the relationship between placenta-related fac-tors and FGR, such as immunization, angiogenesis and apoptosis.

Objective:To observe the change of growth and development, learning and memory, and oxidative stress in serum of offspring rats with fluorosis, and to explore the mechanism of fluoride on neurobehavioral development of offspring rats.Methods:Seventy-two SD rats (female and male ratio 3 ∶ 1) were fed adaptively for one week. According to their body mass [(80 ± 20) g], they were divided into control group (drinking tap water, containing less than 0.5 mg/L fluoride), low fluorine group (drinking water containing 10.0 mg/L of fluoride), and high fluorine group (drinking water containing 100.0 mg/L of fluoride) with random number table. After six months of feeding, they mated freely and gave birth in each group (24 rats with 18 females and 6 males). The rats in each group continued to be exposed to fluoride after giving birth, and the offspring rats were exposed to fluoride through breast milk feeding until the 28th day after birth. Body and brain weight, growth and development indicators (auricle separation, eyes opening, teeth eruption and hair growth) and neurobehavioral development indicators (cliff avoidance, auditory startle, surface righting and vibrissa positioning) were recorded. On the 28th day after birth, the learning and memory abilities (escape latency) of offspring rats were tested by Morris water maze; blood samples were taken from eyeballs to detect the content of nitric oxide (NO), the activity of nitric oxide synthase (NOS) and inducible nitric oxide synthase (iNOS).Results:On the 21st day and 28th day after birth, the differences of body weight among control group, low fluorine group and high fluorine group [21st day: (54.70 ± 3.02), (52.30 ± 2.58), (51.30 ± 2.71) g, 28th day: (91.70 ± 5.03), (90.40 ± 4.76), (86.00 ± 4.55) g] were statistically significant ( F = 3.96, 3.70, P < 0.05); on the 21st day, the body weight of high fluorine group was lower than that of control group ( P < 0.05); on the 28th day, the body weight of the high fluorine group was lower than those of control group and low fluorine group ( P < 0.05). On the 28th day, the difference of brain weight of control group, low fluorine group and high fluorine group was statistically significant ( F = 6.19, P < 0.05); and the low fluorine group and high fluorine group were lower than that of control group ( P < 0.05). Among the growth development indicators, the difference of time of completing eyes opening in control group, low fluorine group and high fluorine group was statistically significant ( F = 3.64, P < 0.05); and the high fluorine group was higher than that of control group ( P < 0.05). In neurobehavioral development indicators, the differences of time of completing cliff avoidance, surface righting between the control group, low fluorine group and high fluorine group were statistically significant ( F = 8.29, 7.69, P < 0.05); and the time of completing cliff avoidance in high fluorine group was higher than those of control group and low fluorine group ( P < 0.05), the time of completing surface righting was higher than that of control group ( P < 0.05). In Morris water maze, on the 4th day, the escape latencies of low fluorine group and high fluorine group were higher than that of control group ( P < 0.05). The results of oxidative stress in serum showed that there were statistically significant differences in serum NO content, NOS and iNOS activitives between the control group, low fluorine group and high fluorine group ( F = 4.86, 66.48, 70.95, P < 0.05); and the NO content of the high fluoirne group was higher than that of the control group ( P < 0.05), the activities of NOS and iNOS of the high fluoirne group were higher than those of control group and the low fluorine group ( P < 0.05). Conclusion:Excessive fluoride can increase the level of oxidative stress in serum, which may be closely related to the neurobehavioral retardation and the decline of learning and memory ability of offspring rats.

Objective:To observe the changes of phosphorylated N-methyl-D-aspartate receptor (P-NMDAR) subunit and calmodulin-dependent protein kinase Ⅱ (CaMK Ⅱ) protein expression in the brain tissue of rats with chronic fluorosis, and to explore the molecular pathogenesis of chronic fluorosis nerve injury.Methods:Eighteen one-month-old SD rats (half male and half female), weighing (100 ± 20) g, were randomly divided into three groups after adaptive feeding for 1 week: control group (drinking tap water, fluoride content < 0.5 mg/L), low fluoride group (drinking water fluoride content was 10.0 mg/L) and high fluoride group (drinking water fluoride content was 100.0 mg/L), with 6 rats in each group (half male and half female). Brain tissue was harvested after 180 days of feeding. HE staining was used to observe the morphological changes of rat brain tissue, Nissl staining was used to observe the changes of Nissl bodies in nerve cells, and immunohistochemical staining was used to observe the expressions of P-NMDAR subunits (P-NMDAR1, P-NMDAR2A) and CaMK Ⅱ protein in rat brain tissue.Results:Under light microscope, HE staining showed disordered arrangement of hippocampal neurons, nuclear hyperstaining and basophilic enhancement in the high fluoride group. The results of Nissl staining showed that the average optical density of Nissl bodies in nerve cells in the hippocampus of rats in the control group, low fluoride group, and high fluoride group were 0.024 4 ± 0.009 7, 0.024 0 ± 0.003 1, and 0.023 9 ± 0.013 8, respectively. There was no statistically significant difference between the groups ( F = 0.010, P > 0.05). Compared with the control group (0.011 8 ± 0.006 5, 0.065 6 ± 0.011 1, 0.143 8 ± 0.029 9) and low fluoride group (0.017 2 ± 0.006 8, 0.062 6 ± 0.017 8, 0.135 6 ± 0.029 6), the protein expressions of P-NMDAR1, P-NMDAR2A and CaMK Ⅱ in high fluoride group were significantly increased (0.026 3 ± 0.005 7, 0.086 3 ± 0.009 0, 0.210 9 ± 0.048 7, P < 0.05); and the protein expression of P-NMDAR1 in low fluoride group was higher than that in the control group ( P < 0.05). Conclusion:Long-term excessive fluoride intake can lead to nerve cell injury in rats, and the mechanism of injury maybe related to the excitotoxicity induced by calcium ion (Ca 2+) overload caused by overactivation of NMDAR subunits.

Objective:To investigate the expression and significance of endoplasmic reticulum stress apoptosis pathway related proteins in renal cortex of rats with chronic fluorosis.Methods:Twenty four healthy SD rats were divided into 4 groups (6 rats/group, half male and half female) according to their body mass (100 - 120 g) by random number table method, rats in control group drank tap water (fluoride content < 0.5 mg/L), and in low, medium and high fluoride groups drank tap water with fluoride content (sodium fluoride) of 10, 50 and 100 mg/L, respectively. After 180 days of feeding, dental fluorosis was examined, 24-hour urine sample was collected and the content of fluoride in urine was detected by fluoride ion selective electrode method. Renal tissue was taken after anesthesia, and the pathological changes of renal cortex were observed by hematoxylin-eosin (HE) staining. The expressions of endoplasmic reticulum stress apoptosis pathway related proteins [inositol-requiring enzyme 1α (IRE1α), apoptosis signal-regulating kinase 1 (ASK1), C-Jun N-terminal kinase (JNK) and phosphorylated JNK (P-JNK)] were determined by immunohistochemical staining in rat renal cortex.Results:No dental fluorosis was found in control group. The incidence of dental fluorosis in low, medium and high fluoride groups were 2/6, 5/6 and 6/6, respectively. Compared with control group [(5.707 ± 1.190) mg/L], the urinary fluoride in low, medium and high fluoride groups [(17.028 ± 3.006), (34.378 ± 12.045), (94.759 ± 31.773) mg/L] was significantly higher ( P < 0.05), and the urinary fluoride in high fluoride group was higher than that in low and medium fluoride groups ( P < 0.05). HE staining showed that, compared with control group, the cell volume of renal tubules and glomeruli in medium and high fluoride groups increased, the cells arranged closely, and the eosinophilia of the cytoplasm increased. The immunohistochemical staining results showed that there was no significant difference in the expression of JNK protein in rat renal cortex between control group and low, medium and high fluoride groups ( F = 0.07, P > 0.05). The expressions of IRE1α, ASK1 and P-JNK proteins in rat renal cortex in high fluoride group were higher than those of control, low and medium fluoride groups ( P < 0.05), and the expressions of IRE1α and ASK1 proteins in medium fluoride group were significantly higher than those in control and low fluoride groups ( P < 0.05). Conclusion:Long-term excessive fluoride intake can lead to renal cortex injury in rats, and the mechanism of injury may be related to the activation of IRE1α-ASK1-JNK endoplasmic reticulum stress apoptosis pathway.

BACKGROUND:Previous studies have shown that N-methyl-D-aspartic acid receptor(NMDA)receptors are associated with fluorine,but the role in fluoride-induced endoplasmic reticulum stress remains unclear. OBJECTIVE:To observe the changes of excitatory neurotransmitter NMDA receptor and endoplasmic reticulum stress IRE1α-ASK1-JNK pathway protein expression in brain tissue of rats with experimental fluorosis,and to investigate the pathogenesis of neurological injury in fluorosis by giving NMDA receptor inhibitor to SH-SY5Y cells. METHODS:(1)Animal model:18 1-month-old SD rats were randomly divided into control group(drinking water fluoride content<0.5 mg/L),low fluoride group(drinking water fluoride content 10.0 mg/L)and high fluoride group(drinking water fluoride content 100.0 mg/L),with 6 rats in each group,half of each sex.After 6 months of fluoride intake,the rats were observed for the occurrence of dental fluorosis,and the 24-hour urinary fluoride content was measured.After anesthesia and euthanasia,the brain tissue of rats was taken to observe the pathological changes.Western blot assay was used to detect NMDA receptors and IRE1α,ASK1 and JNK protein expression in the brain tissue.(2)Cell model:SH-SY5Y cells were cultured in vitro and treated with sodium fluoride at final concentrations of 0.3 mmol/L and 3 mmol/L.The fluoride-stained cells were interfered with 10 μmol/L NMDA receptor antagonists Ifenprodil and MK-801 to observe the relevant protein changes. RESULTS AND CONCLUSION:(1)The incidence of dental fluorosis and urinary fluoride level in rats in the high fluoride group were significantly higher than that in the control and low fluoride groups(P<0.05).(2)Compared with the control group,the cytoplasm of neuronal cells in the CA3 area of the hippocampus in the low fluoride group was slightly more basophilic,while the neuronal cells in the CA3 area of the high fluoride group were disorganized,with increased basophilicity and some of the nuclei solidified.(3)In rat brain tissue,the expressions of NR2A in the high fluoride group and NR2B in the low fluoride group were significantly higher compared with the control group(P<0.05),and NR2B,IRE1,ASK1,and p-JNK protein expression levels were increased in the high fluoride group compared with the control and low fluoride groups(P<0.05).(4)In SH-SY5Y cells,NR1,NR2A and NR2B protein expressions were significantly increased in the high fluoride group compared to the control group(P<0.05).The protein levels of NR1 and NR2A were significantly reduced in the high fluorine + Ifenprodil group and high fluorine + MK-801 group compared with the high fluorine group(P<0.05).NR2B protein expression was significantly lower in the high fluorine + Ifenprodil group than that in the high fluorine group(P<0.05).(5)In SH-SY5Y cells,IRE1,ASK1,and p-JNK protein expression was significantly higher in the high fluoride group compared with the control group(P<0.05),while ASK1 and p-JNK protein expressions were significantly decreased in the high fluorine + Ifenprodil group and high fluorine + MK-801 group compared with the high fluorine group(P<0.05).IRE1 protein level was significantly lower in the high fluorine + Ifenprodil group than that in the high fluorine group(P<0.05).(6)It is concluded that excessive fluorine intake activates NMDA receptors in the central nervous system,causing increased expression of endoplasmic reticulum stress IRE1α,ASK1,and p-JNK proteins,and the use of NMDA receptor inhibitors has a mitigating effect on endoplasmic reticulum stress caused by fluorosis.