Objective To explore the effects of oxidative stress and pulmonary vascular permeability on lung injury in rats treated with sodium cyanide(NaCN) and /or hypobaric hypoxia.Methods A total of 72 male SD rats were randomly divided into 2 groups: the NaCN intoxicated group in 308m altitude and the hypobaric hypoxia combined NaCN intoxicated group in 4 000m high altitude.The animals for the experiment of hypoxia were processed in an artificial hypobaric chamber to simulate the designated high altitude hypoxia(4 000m,61kPa).NaCN was injected subcutaneously to the rats in the both groups at the dosage of 3.6mg/kg.Broncho-alveolar lavage fluid(BALF) and lung tissues were prepared at the time points of 0,0.5,1.0,2.0,4.0 and 6.0h.The activity of SOD,GSH-PX,ACE,LDH,AKP and the content of MDA,GSH,Evan's blue(EB) were detected by spectrophotometric method.Results Combined acute hypobaric hypoxia and NaCN intoxication produced a significant increasing effect on EB content in the rats.The activity of ACE,LDH and AKP in BALF,and the contents of MDA in BALF and lung tissue reached the highest value at 0.5h preparation at the combined hypobaric hypoxia and NaCN intoxicated group,but they were still higher than that of control group(NaCN intoxicated only);correspondingly the activity of SOD,GSH-PX,content of GSH in lung tissue and BALF reached the lowest value at 0.5h preparation at the combined hypobaric hypoxia and NaCN intoxicated group,but they were still lower than that of control NaCN intoxicated group.Conclusion The results suggest that under the condition of hypoxia,NaCN intoxication may produce severe harmful effects on the lungs,increase the pulmonary vascular permeability,and cause severe interference on to the oxidative stress level.

Objective: To verify whether there exists melatoni n(Mel) receptor in human embryonic nervous system. Methods: Spec ific binding of Mel to embryonic brain and spinal cord was measured by radioliga nd binding assay. Results: 125　I-Mel binding s ites in optomeninx was the most, in eptochiasm and sniff ball was next; GTPγS d ose-de pendently inhibited the binding. Conclusion: The results demonst rate the presence of specific binding of Mel in human embryonic brain and spinal cord. GTPγS has some effect on 125　I-Mel specific binding,support ing the theory that Mel receptor is coupled to inhibitory G-proteins.

Objective To compare the outcomes of selective feticide by umbilical cord ligation (UCL),bipolar cord coagulation (BCC) and radiofrequency ablation (RFA) in the treatment of complicated monochorionic twins.Methods We retrospectively analyzed all cases of complicated monochorionic twin pregnancies treated at Peking University Third Hospital from August 2008 to December 2014.The indications for surgery included severe twin-to-twin transfusion syndrome (TTTS),selective intrauterine growth restriction (sIUGR) (type Ⅱ and Ⅲ),twin reversed arterial perfusion sequence (TRAP) or discordant anomaly.One-way ANOVA,LSD t test,Mann-Whitney U test,Chi-square or Fisher's exact test were used for statistical analysis.Results (1) A total of 68 patients chose selective feticide by different techniques,including fetoscopic UCL (n=18,UCL group) and ultrasound-guided RFA (n=46,RFA group).The other four patients treated by bipolar cord coagulation (BCC) were excluded.The maternal age,proportion of assisted reproductive technology,indications,gestational age and mean birth weight all showed no differences between the two groups (P＞0.05).One case of anterior placenta was found in UCL group,fewer than in the RFA group (27 cases,36.9％)(x2=4.853).No fetal loss occurred within two weeks in UCL group,but there were seven cases (seven cases,15.2％) of earlier fetal loss in RFA group (x2=4.952).The median operation time was (63.2±22.5) min in UCL group,and longer than in the RFA group (33.3 ± 11.4) min (t=5.165),all P＜0.05.(2) The gestational age of TTTS and TRAP patients for feticide was older than patients with sIUGR and discordant anomaly [(22.7± 3.0),(22.8±3.2),(20.3 ± 2.5) and (20.4± 3.6) weeks,respectively,F=2.957,P=0.040].Fetal loss rate within two weeks in patients with discordant anomaly was higher than in other groups (4/11 vs 1/10,0/23 and 1/15,P＜0.05).The survival rate,gestational age at delivery and mean birth weight showed no significant differences among the four groups.(3)Compared with feticided fetuses at the upper uterine cavity,the fetal loss rate was higher,and the operation time,gestational age at delivery,birth weight and neonatal survival rate were lower than those performed at the lower uterine cavity,but the difference was not significant.Conclusions RFA provides similar outcomes of selective feticide in complicated monochorionic twins compared with UCL,while RFA is easier to operate.

Objective To evaluate the placental characteristics in monochorionic (MC) twin pregnancy with selective fetal growth restriction (sFGR). Methods Fifty-five placentas from women with MC twin pregnancy were included, who had terminated pregnancy in the Peking University Third Hospital between June 1, 2013 and June 1, 2014, including 23 cases with sFGR and 32 uncomplicated cases as control group. We perfused the placentas within 24 h after delivery, and pigment of four different colors was used to perfuse the umbilical arteries and veins of both twins and determine the types of vascular anastomosis. Umbilical cord insertion, placental territory discordance (PTD, the territory difference between two placentas/the bigger one), and the type, number and diameter of placental superficial vascular anastomosis were analyzed using two independent samples t-test, nonparametric test,χ2 test or Fisher's exact test. Results The PTD was 0.60(0.10-0.80) vs 0.22(0.00-0.90) in sFGR group and control group (Z=-3.913) respectively, and the proportion of placenta with uneven share was 91.3%(21/23) vs 50.0%(16/32) (Fisher's exact test), which were significantly higher in sFGR group (both P < 0.01). The proportion of non-central cord insertion was 82.6% (19/23), 13.0% (3/23) and 40.6% (26/64), respectively, in smaller fetus of sFGR, bigger fetus of sFGR and control group, which was significantly higher in smaller fetus of sFGR than in the other two groups (Fisher's exact test, both P < 0.01). The proportion of arterioarterial (AA), arteriovenous (AV) and venovenous (VV) anastomosis in sFGR group and control group was 78.3%(18/23) vs 75.0%(24/32), 82.6%(19/23) vs 71.9%(23/32), and 17.4%(4/23) vs 15.6%(5/32);there were no significant differences between two groups (Fisher's exact test,all P>0.05). The number of AA, AV and VV anastomosis in sFGR group and control group was 1.0 (0.0-2.0) vs 1.0 (0.0-4.0), 3.0 (0.0-10.0) vs 2.0 (0.0-5.0), and 0.0 (0.0-1.0) vs 0.0 (0.0-3.0) (Z=-0.256, -0.142 and -0.123);the total diameter of AA, AV and VV anastomosis was 2.7 (0.0-7.0) vs 2.2 (0.0-9.7), 4.0 (0.0-13.7) vs 3.4 (0.0-11.5), and 0.0 (0.0-7.9) vs 0.0 (0.0-7.1) mm (Z=-0.070, -0.087 and -0.087);there were no significant differences between two groups (all P>0.05). The total number of all anastomosis was 3.5 (0.0-10.0) vs 3.5 (0.0-6.0) (Z= - 0.567); the total diameter of all anastomosis was 6.9 (0.0-22.4) vs 5.9 (0.0-17.1) mm (Z= - 0.556); there were no significant differences between two groups (all P>0.05). Conclusions Placental sharing discordance and non-central cord insertion may be the risk factors for MC pregnancies complicated with sFGR.

BACKGROUND: Basic fibroblast growth factor (bFGF), a kind of polypeptide growth factor possessing multifunctional biological activities,can protect neurons and promote the growth of nerves. It has been corfirmed that bFGF has therapeutic effects on retina ischemia/reperfusion injury (RIRI).OBJECTIVE: To establish RIRI model and analyze the effects of bFGF on cellular apoptosis of retina and the expression of regulatory gene protein.DESIGN: Randomized grouping and validating trial.SETTING: Department of Ophthalmology, the Affiliated Hospital of Medical College of Qingdao University.MATERIALS: The experiment was conducted at the Research Laboratory of Pathology, Department of Ophthalmology, Medical College of Qingdao University, from April 2002 to December 2003. Twenty-eight healthy Wistar rats were enrolled in this experiment. Four rats were randomly chosen for normal control group, the left eyes of the other 24 rats were set as normal saline control group, and the right eyes were set as bFGF group.METHODS: Normal saline control group and bFGF group adopted the rat RIRI models established by transiently elevating intraocular pressure. Normal saline of 12 μL was injected into the vitreous cavity of the left eyes of the rats in normal control group. 12 μL bFGF was injected into the vitreous cavity of the right eyes of the rats in bFGF group, 4 rats once. No administration was given in normal control group. The expression of apoptotic cells was detected and apoptosis indexes were calculated with the terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick-end labeling (TUNEL) method and immunohistochemical staining method at the 1st, 6th,12th, 24th,48th and 72nd hours after reperfusion and ischemia for 1 hour.MAIN OUTCOME MEASURES: ① The detection results of apoptotic cells in situ of retina tissuesat different time points after reperfusion. ②The expression of Fas and caspases-2 in retina tissues at different time points after reperfusion.RESULTS ① Comparison of apoptosis indexes of retina tissues at different time points after ischemia reperfusion: There were no apoptotic cells in the retina tissues of the rats in normal control group. As compared with those in normal saline control group, apoptosis indexes in bFGF group were significantly decreased at ischemia 1 hour and reperfusion 1, 6, 12, 24, 48and 72 hours, especially at the 12th, 24th and 48th hours after reperfusion (t =5.362-5.595, P ＜ 0.05). ② The change of Fas expression at different time points after ischemia reperfusion: There was hardly any Fas expression in normal control group. As compared with that in normal saline control group, Fas expression in bFGF group was significantlydecreased at ischemia 1 hour and reperfusion 1, 6, 12, 24, 48 and 72 hours, especially at the 6th, 12th and 24th hours after reperfusion (t=3.954-9.327, P ＜ 0.05). ③The changes of caspase-2 expression at different time points after ischemia reperfusion: There was no caspase-2 expression in normal control group.Compared with that in normal saline control group, the number of caspase2 positive cells in bFGF group was significantly decreased at the 6th,12th,24th, 48th and 72nd hours after ischemia for 1 hour and reperfusion (t=4.125-15.641, P ＜ 0.05).CONCLUSION: bFGF can significantly inhibit the expression of apoptosis gene Fas and caspase-2 in the ischemia and reperfusion of retina, thus reducing cellular apoptosis of ganglion cells and exerting therapeutic effects on the ischemia and reperfusion of retina.

Background Acanthamoeba keratitis is a sort of serious infectious eye disease with high causing-blindness rate.Acanthamoeba keratitis often is misdiagnosed as fungal keratitis or viral keratitis in the early stage.Because conventional clinical diagnosis methods show a low specificity and take a long time,timely treatment often is delayed.Conventional PCR does not apply well because the lesion sample is not enough to extract DNA.However,direct PCR can amplify 18S rRNA conserved sequence of acanthamoeba keratitis without the extraction of DNA.Objective This study was to discuss the feasibility for rapid diagnosis of acanthamoeba keratitis using direct PCR to amplify the gene 18S rRNA fragment.Methods Ten acanthamoeba strains were isolated from 10 eyes with acanthamoeba keratitis in Qingdao Eye Hospital.The sensitivity of the direct PCR assay was tested using different numbers of amoebas.The specificity of the assay was tested using DNA extracted from acanthamoeba,candida albicans,pseudomonas aeruginosa,herpes simplex virus-1 (HSV-1) and normal human corneal epithelial cell.Acanthamoeba keratitis models were established using infected method in clean 6-week-old female BALB/c mice.Corneal lesion samples were obtained 1 day,3,5,7,10,15 days after modeled.The effectivity and feasibility of the direct PCR assay for rapid diagnosis of acanthamoeba keratitis were evaluated and compared with culture method,corneal smear examination and real-time PCR.Results Direct PCR primers could only amplify DNA of acanthamoeba rather than other pathogens,and 10 stains of acanthamoeba were detected at least in each sample.During the development of acanthamoeba keratitis in the mice,the diagnosis positive rate of direct PCR was 80.0％,90.0％,80.0％,70.0％,70.0％ and 50.0％ in 1 day,3,5,7,10,15 days after modeled with the total positive rate 73.3％,which was higher than 31.7％ of culture method,56.7％ of corneal smear examination and 61.7％ of realtime PCR,with a significant difference between the direct PCR and culture method (P =0.005),but no significant difference was seen in the total positive rate between the direct PCR and real-time PC R (P =0.172) or corneal smear examination (P =0.056).Conclusions The direct PCR assay is a simple,rapid,highly specific and sensitive method for the rapid diagnosis of acanthamoeba keratitis,especially for the limited lesion sample.

Objective To describe and analyze the clinical characters of patients with FRG from 7 Chinese families.Then analyze and identify their mutations in SGLT2 gene,and explore the association of genotype and phenotype.Methods Quantitative test for 24-hour urine glucose and other laboratory tests were carried out among 7 probands (14 patients in all) and their family members from 7 pedigrees (totaling 23 subjects).All coding regions,including intron-exon boundaries,were analyzed using PCR followed by direct sequence analysis.Results Five novel mutations in SLC5A2 gene were identified in this investigation,including four missense mutations (A Serine to Glycine at position 335 (c.1003A＞G,p.S335G),a Glutamine to Arginine at position 448 (c.1343A ＞ G,p.Q448R),an alanine to proline at position 474 (p.A474P,c.1420G ＞ C) and a glycine to aspartic acid at position 580 (c.1739G ＞ A,p.G580D) and a deletion in intron 7 (c.886(-10_-31)del).By the minigene studies using the pSPL3 plasmids,we confirmed the deletion c.886(-10_-31)del as a splicing mutation.In this study,the mutation c.886(-10_-31)del accounted for about 43％ of the total alleles (12/28).These patients with compound heterozygous or homozygous mutations manifested middle degree or severe glycosuria (Quantitative test for 24-hour urine glucose:10.56-50.68 g/1.73 m2),however those with heterozygous variants presented with mild to moderate glycosuria (Quantitative test for 24-hour urine glucose ≤ 2.45 g/1.73 m2).This fits co-dominant inheritance pattern.Conclusions Five novel mutations which may be related to FRG are found in this study,and c.886(-10-31) del may be a high frequency mutation in Chinese patients.

Objective:To select suitable chemotherapy for cervical cancer patients by ATP-tumor chemosensitivity assay.
Methods:Seventy-two hospitalized patients with cervical cancer between July 2007 and October 2009 were enrolled. The patients were randomly divided into a trial group (n=35) and a control group(n=37). ATP-TCA was used to detect the sensitivity of 35 samples of cervical cancer in the trial group to 6 combined chemotherapy regimens. The chemotherapy regimen in the trial group was confirmed by the results of susceptibility testing and that in the control group was confirmed by clinical experience. One-year recurrence rate and 3-year survival rate of two groups were compared after 3 year follow-up.
Results:ATP-TCA was measured in 32 of the 35 patients in the trial group. The sensitive patients for paclitaxel+carboplatin, paclitaxel+oxaliplatin, bleomycin+ifosfamide+cisplatin, bleomycin+vincristine+cisplatin, fluorouracil+cisplatin, and gemcitabine+cisplatin were 20, 18, 17, 18, 17, and 21, respectively. There was no significant difference in the 1-year recurrence between the two groups (P>0.05), while the 3-year survivors in the trial group were more than those in the control group (P<0.05).
Conclusion:ATP-TCA method is good for patients with cervical cancer because it is sensitive, effective, and individualized.

Objective In this study,maximum platelet aggregation rate of Light transmittance aggregometry (LTA) for coronary heart disease(CHD) patients taking antiplatelet drug and patients without antiplatelet therapy was measured in non-adjusted and platelet count-adjusted platelet-rich plasma (PRP).The aim of this study is to compare which method is superior in evaluation of antiplatelet drug effect.Methods This is a methodology comparative research.560 CHD outpatients and inpatients that visited Beijing Anzhen Hospital of the Capital University of Medical Sciences from May to June,2012 were chosen,who were treated with aspirin monotherapy,or patients on combination therapy with aspirin and clopidogrel,as well as patients without antiplatelet therapy.LTA was performed in non-adjusted (improved method) and platelet count (200 × 109/L)-adjusted PRP (original method),using 6 μmol/L adenosine diphosphate (ADP) and 0.5 mmol/L arachidonic acid (ARA) as agonists.The maximum aggregation rates in 5 min were detected,and consistency and differences of the two methods were compared.Results There is no statistically significant correlation between maximum aggregation rate and platelet count in PRP with 6 μmol/L ADP or 0.5 mmol/L ARA as agonists in all subgroups including aspirin monotherapy,combination therapy with aspirin and clopidogrel and patients without antiplatelet therapy (-0.21 ≤ r ≤0.111,P ＞ 0.05).The maximum aggregation rate using ADP as agonists in original method is decreased compared with improved method,there is statistically significant difference in all subgroups including patients without antiplatelet therapy,aspirin monotherapy,combination therapy with aspirin and clopidogrel less than one week and more than one week.The variability of platelet aggregation rate using ADP as agonists with improved method is lower than that with original method in all subgroups.Yet the maximum aggregation rates using ADP as agonists with improved method and original method correlate well with each other in all subgroups (r =0.78,0.73,0.40,0.71,P ＜0.01).In the subgroup of subjects without antiplatelet therapy using ARA as agonist,platelet aggregation rate is decreased in original method compared with improved method,there is statistically significant difference,and the variability of the aggregation rate with improved method is also lower than that with original method,ranging from 62％-98％ relative to 5％-89％.The decrease of aggregation rate using ARA as agonist for patients taking antiplatelet drug compared with patients without antiplatelet therapy can be detected both with improved method and original method.Conclusion Non-adjusted PRP in LTA is more convenient and time-saving,and it also means less effects on platelet in vitro.Therefore,non-adjusted PRP is more suitable for monitoring efficacy of antiplatelet therapy in clinical laboratory.

In many pathogens infection,especially virus,antibody-dependent enhancement(ADE) can aggravate the infection and lead to severe diseases.In this immunopathological phenomenon,virus-specific antibodies enhance the entry of virus into monocytes,macrophages and granulocytic cells and even the replication of virus through different mechanism.This phenomenon has been reported in numerous pathogens including virus,bacteria and parasite and the mechanisms of ADE vary from different species.Further study of ADE can promote the vaccine research and development to make the most use of vaccine and prevent human body from pathogens,which will be helpful to control the spread of pathogens including Zika virus.In the present review,we review the research progress of ADE mechanism in recent years,including antibodies mediating,receptors mediating,complement mediating,viral proteins mediating and cellular mediating ADE.In addition,dengue virus,human immunodeficiency virus,Coxsackie virus,Ebola virus,Zika virus and other pathogens will be illustrated respectively.This review provides insights on the different mechanism of ADE in different pathogens.