Objective To observe the effects of butylphthalide on the expression of autophagy-related protein and mRNA in l-meth-yl-4-phenyl-pyridiniumion (MPP+)-induced SH-SY5Y cells, and to explore the protective effect and possible mechanism of butylphthalide to the cell model of Parkinson's disease. Methods The SH-SY5Y cells were divided into control group (A), MPP+group (B), rapamycin pre-treated+MPP+group (C) and Butylphthalide pretreated+MPP+group (D). The relative viability of SH-SY5Y cells induced by MPP+was measured with MTT assay, the morphology of SH-SY5Y cells was observed. The expression of microtubule associated protein 1 light chain 3 (LC3)-II/I and Beclin 1 protein was detected by Western blotting. And the expression of LC3-II/I and Beclin 1 mRNA were assayed by re-al-time quantitative reverse transcription-PCR (RT-PCR). Results The viability rates of cells were significantly lower in group B than in group A (t=20.270, P<0.001), and were significantly higher in groups C and D than in group B (t>8.770, P<0.001), however, there was no significantly difference between groups C and D (t=2.270, P=0.064). The expression of LC3-II/I and Beclin 1 was higher in group B than in group A (t>6.647, P<0.01), and was higher in groups C and D than in group B (t>3.630, P<0.01), however, there was no significantly differ-ence between groups C and D (t<2.238, P≥0.05). Conclusion Butylphthalide could prevent the injury of SH-SY5Y cells induced by MPP+, which may affect Parkinson's disease by inducing autophagy.

ObjectiveTo survey the status of Helicobacter pylori (HP)infection in cholangiocarcinoma,and its relation to clinical and pathological parameters and prognosis. MethodsHP infection in 80 cholangiocarcinoma samples and 30 controls was detected by PCR,in those with positive results the expression of Cag A and its subtypes of Vac A sla,m1 and m2 was further tested by PCR.ResultsChi-square test showed that the detection rate of in HP ( + ) in cholangiocarcinoma group is 71％,higher than 20％ in control group.The positive cases of CagA and VacA sla,m1,m2 in cholangiocarcinoma group was respectively 30,40,5 and 43 cases.HP infection in cholangiocarcinoma was correlated with the location of the tumor(x2 =27.580,P ＜ 0.05 ). MultivariateLogisticanalysisshowedthat cholangiocarcinoma is over 10 times more likely in HP ( + ) patients than HP ( - ) ( OR =10.531 ).Cox regression analyses showed that the infection of HP(HR =8.105,P =0.032),the staging of TNM( Ⅱ/ⅢHR=9.141,P=0.040,Ⅳ HR =29.071,P=0.040) and surgery (HR=9.531,P =0.015) are all independent prognostic factors of cholangiocarcinoma. Life table analyses showed HP infection negatively affects the survival time of cholangiocarcinoma after a surgery ( u =10.074,P =0.002),and the median survival time is 7.25 months shorter than HP( - ) patients. ConclusionsIt is common that HP infection complicating cholangiocarcinoma,usually with the genotype of VacA sla/m2,HP infection is a risk factor for cholangiocarcinoma,and negatively affects oatients survival after surgery.

Objective To investigate the incidence and clinical presentation of breast milk transmitted cytomegalovirus (CMV) infection among preterm infants with birth weight≤1500 g.Methods Preterm infants enrolled in this study met the following inclusion criteria: birth weight≤1500 g, fed with CMV-positive breast milk and admitted into Neonatal Intensive Care Unit of Children's Hospital of Fudan University within 72 hours after birth from October 2015 to July 2016. And those with congenital digestive tract malformation or congenital CMV infection were excluded. Breast milk and infants' urine samples were regularly screened for CMV DNA by fluorescent quantitative polymerase chain reaction. Symptoms and laboratory findings in infants with CMV infection transmitted via breast milk were documented and analyzed. Differences in relevant parameters were analyzed usingChi-square test, Fisher's exact test,t test or Mann-WhitneyU test where appropriately.Results Sixty preterm infants breastfed with CMV DAN-positive milk were recruited. Among them, 19 (31.7%) developed breast milk-acquired CMV infection as their urine samples were positive for CMV DNA, while the others were negative for CMV DNA (infected group:n=19; non-infected group:n=41). The average CMV copies in breast milk, gestational age and birth weight of the infected group were all significantly higher than those of the non-infected group [3.76 (3.18-4.50) vs 3.47 (3.00-4.88) Log10 copies/ml,Z=-2.042;(30.4±2.1) vs (28.4±2.3) weeks,t=3.175; 1290 (750-1500) vs 1110 (575-1480) g,Z=-2.837; all P<0.05). Fewer infants in the infected group than in the non-infected group received blood transfusion [5/19 vs 56.1%(23/41),χ2=4.627,P<0.05]. Ages of the infants with CMV infection ranged from 26 to 164 days (median age of 92 days). Six out of the 19 infants had clinical symptoms concurrent with viral excretion in urine and the ages of these symptomatic infants of infection were earlier than those of the asymptomatic ones without significance [(72±34) vs (97±28) days,t=-1.710,P>0.05]. Four infants (21.1%, 4/19) had severe organ damage and/or positive IgM antibodies to CMV in serum, and were treated with antiviral therapy. Two had mild symptoms and were not given antiviral therapy. All of the six symptomatic infants were followed-up for one to six months, during which time the complete blood cell count and results of biochemical test and fundus examination were back to normal.Conclusions The incidence of breast milk-acquired CMV infection among preterm infants with birth weight≤ 1500 g was 31.7%, and no severe symptoms were reported in this study.

Aim To observe histopathological changes of hippocampus after acute epilepsy induced by penty-lenetetrazole (PTZ)in rats.Methods Five groups as control group,PTZ-induced 24 hours(h)group,PTZ-induced 72 hours group,PTZ-induced 1 20 hours group and PTZ-induced 1 44 hours group were designed.PTZ (64 mg·kg -1 )was administered with a single intrap-eritoneal injection for generalized tonic-clonic sei-zures in the current experiment.Control and PTZ trea-ted animals were sacrificed after specific time points. Brain was dissected out and then evaluated for neuro-pathological changes using Nissl staining and immuno-histochemical technique.Results In this study PTZ-induced hippocampal neuron status apoptosis occurred at 24 hours and was sustained for 1 44 hours after status epilepticus.Whereas,activated caspase-3 and AIF ap-peared at 24 hours and were sustained for 1 44 hours af-ter status epilepticus.Conclusion The results of this study show that the significant histopathological chan-ges of hippocampus appear in the vicinity of 1 20 hours after intraperitoneal injection of pentylenetetrazole.

Objective To investigate Nuclear factor-κB (NF-κB) expression in tumor associated inflammation in patients with adamantinomatous craniopharyngima..Methods Fifty-four patients (31 male and 23 female) with craniopharyngioma from 3 to 66 years of age were recruited from May 2004 to March 2006.NF-κB and Osteopontin (OPN) expression in human craniopharyngiomas were detected using immunohistochemical staining.High-sensitivity C-reactive protein (hs-CRP), a systemic marker of inflammation, was examined in patients' tumor hydatid fluid, cerebrospinal fluid and serum.Results NF-κB expression was significantly increased in the adamantinomatous craniopharyngioma.Spearman;s correlation analysis demonstrated that NF-κB expression was associated with OPN expression.The hs-CRP level was also increased in the tumor hydatid fluid (4.28±0.90 mg/mL), cerebrospinal fluid (0.035±0.006 mg/mL) and serum (1.72±0.54 mg/mL) in patients with adamantinomatous craniopharyngioma.Conclusions NF-kappa B is closely associated with tumor associated inflammation which further mediates adhesion of tumor to surrounding important structures in patients with adamantinomatous craniopharyngioma.

Objective To investigate and evaluate the correlation between brain natriuretic peptide (BNP) and gestational hypertension and preeclampsia of hypertensive disorder complicating pregnancy (HDCP).Methods Fifty cases with HDCP and 46 cases with mild and 83 cases with sever stage preeclampsia were selected as our subjects.And 33 cases with regular pregnancy and 31 with irregular pregnancy were served as control group.Plasma brain natriuretic peptide,urinary protein quantity(UBQ),24-hour urinary protein assay (UPA) were measured.The correlations of brain natriuretic peptide and UBQ,UPA,systolic pressure (SP),diastolic pressure (DP) were analyzed.Results The levels of brain natriuretic peptide in the group with gestational hypertension and mild,severe preeclampsia groups were (48.54± 18.27),(79.46± 32.18) and (292.24±213.08) ng/L,higher than that in normal pregnancy and non pregnant group ((27.84± 14.58) and (20.63± 8.28) n/L;F =49.583,P＜0.05).While no significant difference exists between normal pregnancy group and non pregnant group.Grouped on the median values (199) of brain natriuretic peptide of the severe preeclampsia group,the levels of 24-hour UPA,systolic pressure and diastolic pressure were (5.46±2.68) g,(174.55± 13.58) mmHg,(113.74±9.91) mmHg in patients with brain natriuretic peptide ≥ 199 ng/L(n=42),significant higher than those in patients with brain natriuretic peptide ＜ 199 ng/L(n =41;(4.34± 1.95)g,(165.31±11.12) mmHg,(106.05±8.02) mmHg;t=2.603,3.396,2.308;P=＞0.010,0.001,0.024).The levels of 24-hour UPA,systolic pressure and diastolic pressure of patients with brain natriuretic peptide ≥ 86ng/L(n=20) in mild preeclampsia were (1.68±0.27) g,(163.69±8.29) mmHg,(105.45±6.71) mmHg,significant higher than those in patients with brain natriuretic peptide ＜ 86 ng/L (n =26;(1.16 ± 0.31) g,(152.90±7.32) mmHg,(99.19 ± 5.25) mmHg;t =3.180,2.508,2.32;P =0.010,0.016,0.025).Brain natriuretic peptide was closely correlated with UPA,systolic pressure and diastolic pressure in hypertensive disorder complicating pregnancy (HDCP) (r =0.29,0.30;P ＜ 0.01).Brain natriuretic peptide was closely correlated with UPA systolic pressure and diastolic pressure in mild preeclampsia (r =0.39,0.37,0.40;P ＜0.01).And correlation efficacy of brain natriuretic peptide with UPA,systolic pressure and diastolic pressure were 0.44,0.42 and 0.53 (P＜0.01).Conclusion The level of brain natriuretic peptide is closely associated with the severity of gestational hypertension and preeclampsia of hypertensive disorder complicating pregnancy.Correlation of brain natriuretic peptide to the severity of gestational hypertension and preeclampsia is independent of urinary protein and hypertension.Brain natriuretic peptide is an important indicator for the severity of gestational hypertension and preeclampsia of hypertensive disorder complicating pregnancy.

Abstract Gram-negative ( G-) bacteria, such as denitrifying bacteria and anaerobic ammonia oxidation bacteria, are highly social organisms capable of sophisticated cooperative behavior mediated via quorum sensing. As signal molecules of the chemical communication, N-acyl-homoserine lactones ( AHLs ) can mediate the quorum sensing of the functional microbial population and regulate the population density. To understand the growth of functional microbial population and the mechanism for biological nitrogen removal in upflow microaerobic sludge reactors ( UMSRs ) treating organic wastewater with low ratio of chemical oxygen demand to total nitrogen, a method was established to simultaneously detect AHLs in the microaerobic processes. Water-sludge mixtures sampled from the UMSRs were pretreated in sequence by liquid-liquid extraction using ethyl acetate, rotary evaporation, constant volume with methanol, separation by C18 column. Gradient elution was carried out using 5 mmol/L ammonium acetate ( containing 0 . 1% formic acid ) and methanol as mobile phases. On the base of multiple reaction monitoring analysis, a triple quadrupole mass spectrometer with an electrospray ionization was introduced to detect the target compounds. Nine kinds of AHLs were used to evaluate the established method and the results showed that the detection limits were 0 . 01-0 . 5 μg/L and all of the AHLs presented excellent linearity with the concentration ranging from 0 . 5 to 100 μg/L. The recovery and relative standard deviation ranged from 62. 5% to 118. 1% and 2. 9% to 12. 1%, respectively. The analysis could be finished within 6. 5 min. The rapid, accurate and precise method for detecting AHLs provided a new insight into the growth and metabolic activity of functional microbial population in the activated sludge processes to understand the mechanism of biological nitrogen removal, suggesting a good application in regulation and operation of wastewater biological treatment processes.

Objective To observe the effects of Jianpi Huoxue Prescription on the subcutaneous transplantation of tumor cell apoptosis and angiogenesis; To discuss its possible mechanism of action. Methods SMMC-7721 hepatoma mice models were established. 60 tumor bearing male mice were randomly divided into blank group, model group, 5-Fu group, Jianpi Huoxue Prescription high-, medium- and low-dosage groups, with 10 rats in each group. Model group and Jianpi Huoxue Prescription high-, medium- and low-dosage groups were given medicine with relevant concentrations for gavage, once a day. 5-Fu group was given 5-Fu for intraperitoneal injection every other day. The weight, tumor weight and tumor inhibitory rate of each group were compared two weeks later. The protein expressions of VEGF and VEGFR-2 were detected by immunohistochemistry, microvessel density (MVD) were calculated, and the expressions of Survivin and Caspase-3 mRNA were detected by RT-qPCR. Results Compared with model group, protein expressions of VEGF and VEGFR-2, MVD, Caspase-3 and Survivin mRNA decreased in Jianpi Huoxue Prescription high-, medium- and low-dosage groups, and the most obvious differences were in 5-Fu group and Jianpi Huoxue Prescription high-dosage group (P<0.05). Conclusion Jianpi Huoxue Prescription can inhibit tumor growth by inhibiting angiogenesis and inducing apoptosis.

Objective To explore the effects of butylphthalide on the apoptosis of SH-SY5Y cells induced by the 1-methyl-4-phenylpyridinium iodide (MPP+). Methods SH-SY5Y cells were cultured with butylphthalide in the dosage of 1 µmol/L, 10 µmol/L and 20 µmol/L for 2 hours, and with MPP+ for 24 hours. The viability of cells was measured with trypan blue. The mitochondrial membrane potential was detected with Rhodamine 123. The content of P53 protein was detected with ELISA. The expression of P53 and cytochrome C (CytC) were detected with Western blotting. Results The viability of SH-SY5Y cells increased with the dosage of butylphthalide (P<0.05), while the mitochondrial membrane potential and the expression of P53 and CytC decreased (P<0.05). Conclusion Butylphthalide can prevents the SH-SY5Y cells from the injury induced by MPP+, which may associate with the inhibition of apoptosis through the expression of P53 and CytC.

Exosomes are a kind of endosomal vesicles that are secreted by most if not all living cells. Due to their capability of delivering a variety of cargos, such as tissue- or cell-specific proteins, lipids, and genetic materials, and their broad biological activities, exosomes have gained substantial attention as emerging therapeutics. Exosomes derived from mesenchymal stem cells (MSCs) and dendritic cells (DCs) are two types of exosomes that are widely studied. Many preclinical and clinical studies have shown that they have a satisfactory treatment effect in lung diseases, liver diseases, nervous system diseases, tumors, and other diseases. In addition, exosomes from macrophages, tumor cells, plant cells, and many other cells are getting more attention due to their therapeutic potential. Besides natural exosomes, research on engineered exosomes has also made plenty of progress. There have been several engineering methods of exosomes, such as targeting modification and loading of active ingredients. In this review, we summarize the research progress of therapeutic exosomes from different sources, and further discusses the application prospects of exosomes and possible challenges in the future.