Objective:To establish and optimize a detection method of dibutyryl cyclic adenosine phosphate related substances.Methods:On the basis of the JP,the experimental conditions were optimized to obtain the optimal detection conditions.The column was CAPCEILPAK C18;mobile phase was pH 6.0 phosphate buffer solution-meth-anol with gradient elution;detection wavelength was 258 mm.The method validation was carried out.Results:In the system applicability solution,the separation degree of the main peak and adjacent peaks in each destruction solution was greater than 1.5;the solution was stable within 48 h.The content of each impurity showed a good line-ar relationship with the peak area in the range of 50％-150％of the limit concentration,R2＞0.999 4.In the spike experiment,the average recovery rate of N6-butyryl 3',5-cyclophosphatin sodium was 99.4％-106.6％.The average recoveries of 2'-0 butyryl 3,5-cycloadenosine monophosphate sodium were 88.8％-103.6％,and the average recoveries of cyclophosphaminate sodium were 96.9％-102.5％,which met the requirements.Three bat-ches of samples were tested,and the above impurities were detected.Conclusion:This method has high sensitivi-ty,good separation effect,accuracy and durability,and can be used for the detection of bucladesine sodium,provi-ding a basis for its quality control.

Interferon-γ (IFN-γ), a key effector molecule in anti-tumor immune response, has been well documented to correlate with the intratumoral infiltration of immune cells. Of interest, however, a high level of IFN-γ has been reported in salivary adenoid cystic carcinoma (SACC), which is actually a type of immunologically cold cancer with few infiltrated immune cells. Investigating the functional significance of IFN-γ in SACC would help to explain such a paradoxical phenomenon. In the present study, we revealed that, compared to oral squamous cell carcinoma cells (a type of immunologically hot cancer), SACC cells were less sensitive to the growth-inhibition effect of IFN-γ. Moreover, the migration and invasion abilities of SACC cells were obviously enhanced upon IFN-γ treatment. In addition, our results revealed that exposure to IFN-γ significantly up-regulated the level of programmed death ligand 1 (PD-L1) on SACC cell-derived small extracellular vesicles (sEVs), which subsequently induced the apoptosis of CD8⁺ T cells through antagonizing PD-1. Importantly, it was also found that SACC patients with higher levels of plasma IFN-γ also had higher levels of circulating sEVs that carried PD-L1 on their surface. Our study unveils a mechanism that IFN-γ induces immunosuppression in SACC via sEV PD-L1, which would account for the scarce immune cell infiltration and insensitivity to immunotherapy.
B7-H1 Antigen/metabolism* ; CD8-Positive T-Lymphocytes/pathology* ; Carcinoma, Adenoid Cystic/pathology* ; Carcinoma, Squamous Cell/pathology* ; Cell Line, Tumor ; Humans ; Immunosuppression Therapy ; Interferon-gamma/pharmacology* ; Mouth Neoplasms/metabolism* ; Programmed Cell Death 1 Receptor/metabolism* ; Salivary Gland Neoplasms/pathology*

OBJECTIVE：To investigate the accessibility of antibiotics in essential medicine list in medical institutions of Nanjing area ，and to provide reference for formulating and improving related drug policy. METHODS ：Taking the original and imitated drugs as the objects of investigation ，the standard investigation method jointly formulated by WHO/HAI was adopted to select 35 kinds of antibiotics which were included in Essential Medicine List （2018 edition）and International Drug Price Indicator Guide（2015 edition），and to investigate 3 dimenions such as the availability （proportion of institutions that could provide this drug），price（calculated by MPR ）and affordability （the ratio of drug expenditure to the daily minimum wage of non-technical government workers ）of them in 13 second-class or above medical institutions in Nanjing area by online questionnaires during Jan.-Jun. 2019，so as to put forward the suggestions for drug policy formulation. RESULTS ：The median availability of original drugs was 0，with a range of 0 to 100％；the median availability of generic drugs was 30.80％，with a range of 0 to 100％. The median MPR of original drugs was 5.54，with a range of 1.96 to 18.83；the median MPR of generic drugs was 1.76，with a range of 0.16 to 7.66. Median affordability of original drugs was 8.68，with a range of 1.19 to 41.79；the median affordability of generic drugs was 0.52， with a range of 0.03 to 16.80. CONCLUSIONS：Generic drugs are more available than the originals. The price of original drugs is generally very high ， while the price of generic drugs is mostly cheaper. The affordability of original drugs is really poor ， and the 规。E-mail：shaorong118@163.com affordability of generic drugs is totally good. There are stillrooms for further improvement in the overall accessibility of essential medicines. At present ，ensuring the accessibility of original drugs may be more helpful to meet the needs of treatment. It is suggested to update and adjust the essential medicine list based on the clinical medication requirement ，popularize the knowledge about essential medicines ，properly adjust the price of original drugs and ease the treatment burden of patient. It is supposed to take into full consideration about the regional factors and the needs of different medical institutions when making drug policy.

Objective To investigate the mental states of the community-dwelling elderly in Beijing ,and to explore their relationship with depression using multivariate analysis ,in order to provide evidence for strategies to prevent depression in the elderly. Methods A total of 400 community elderly residents in Chaoyang District of Beijing were recruited in this cross-sectional study. The mental states of older adults were assessed by using the Zung Self-Rating Depression Scale(SDS) ,the Medical Outcomes Study(Mos)36-item Short Form Health Survey (SF-36) ,and the Social Support Rating Scale(SSRS).The correlations of the mental state with demographic data ,quality of life and social support were examined via multivariate analysis. Results Of 400 questionnaires ,364 were valid.Depression occurred in 64 older adults ,and the detection rate was 17.6% (64/364).Multivariate linear regression analysis showed that depression was closely correlated with age ( P = 0.049 ) , physiological function(P=0.009) ,vitality(P=0.028) ,mental health(P＜0.001) ,self-reported health transition(P=0.003) ,objective support(P=0.001)and subjective support(P=0.001)in community-dwelling elderly residents in Beijing.Bivariate Logistic regression analysis showed that five factors , including vitality (OR = 0.595 ,P= 0.036 ) ,mental health (OR = 0.548 ,P = 0.021 ) ,self-reported health transition(OR=1.597 ,P= 0.048) ,objective support (OR= 0.524 ,P= 0.018)and subjective support(OR=1.787 ,P=0.027) ,were influencing factors for depression in the elderly.Self-reported health transition and subjective support had the greatest impact on depression in the elderly . Conclusions Critical approaches to preventing depression in the elderly include a focus on the mental health and social needs as well as measures to enhance self-confidence in the health of these individuals.

Objective To determine whether the antiserum produced by immunizing mice with conotoxin GI coupled with bovine serum albumin (BSA) could neutralize GI conotoxin.Methods The GI-BSA was prepared by glutaraldehyde-coupled method,and the mice were immunized with the GI-BSA to produce antiserum.The antibody neutralization assay was used to test the detoxication of the antiserum.Results The SDS-PAGE protein electrophoresis showed that the coupling reaction of GI hapten with BSA was successful.The two distinct protein bands of GI-BSA were more than 120×103.Each mouse was immunized four times with 99 μg every two weeks.After the fourth immunization,the serum neutralization titer was more than 1:64 000.After the intraperitoneal injection of the mixture of 100 or 200 μl of the antiserum and different doses of GI,75% of the mice survived in the group with 100 μl of the antiserum and 1× LD50 GI(16.3 μg/kg).The same percentage of mice also survived in the group of with 200 μl of serum and 25.8 μg/kg of GI.Conclusion The antiserum produced by immunizing mice with GI-BSA exhibits significant detoxication activity to conotoxin GI.

ObjectiveTo study the effects of group BStreptococcus (GBS) colonization in late pregnancies on neonatal GBS infection.MethodsA total of 17 019 pregnant women who received antenatal care and delivered in Xiamen Maternal and Child Care Hospital from June 1, 2014 to May 31, 2015 were enrolled in this study. Secretions from the lower third of the vagina in the pregnant women at 35-37 weeks of gestation or having premature baby(regardless of gestational age) were obtained to test GBS by standard bacterial culture, and 1 472 cases underwent GBS DNA test by real-time fluorescent quantitative-polymerase chain reaction (PCR) meanwhile. The pregnant women colonized with GBS (GBS culture and/or PCR DNA test positive) were given intrapartum antibiotic prophylaxis (IAP) during parturition or rupture of fetal membranes. Detection rate of the two methods was compared, and the effects of GBS colonization and IAP on neonatal GBS infection were analyzed to identify the risk factors of neonatal early-onset GBS disease (GBS-EOD). Two independent samplest-test,Chi-square test and Logistic regression analysis were used for statistical analysis. ResultsThe detection rate of GBS culture and PCR DNA test was 14.43% (2 456/17 019) and 14.13%(288/1 472), respectively. The total colonization rate was 14.52%(2 472/17 019). Based on the culture results as golden criteria, the sensitivity, specificity, positive predictive value and negative predictive value of PCR assay were 95.05%, 98.74%, 92.31% and 99.21%, respectively. There were 17 332 deliveries from the 17 019 pregnant women, of which 31 cases had GBS-EOD. The incidence of neonatal GBS-EOD in maternal GBS colonization [1.05%(26/2 472)] was 31 times higher than in pregnant women without GBS colonization [0.34‰(5/14 547)]. Among the 31 infants with GBS-EOD, 24 had pneumonia, five had sepsis, and two had meningitis. The case fatality rate was 6.45%(2/31). Logistic regression analysis found that chorioamnionitis was an independent risk factor of neonatal GBS-EOD (OR=40.425, 95%CI: 7.514-379.782,P=0.000). Compared with the non-IAP group,IAP group had a lower incidence of GBS-EOD among the pregnant women colonized with GBS [0.94%(23/2 443) vs 10.34%(3/29),χ2=24.350,P<0.01].ConclusionsGBS colonization in late pregnant women has adverse effects. Therefore, routine maternal rectovaginal culture of GBS may be necessary and IAP should be applied in those with GBS colonization.

ObjectiveTo dynamically observe the effects ofJieduan Niwan Formula on mitochondrial apoptotic pathway in acute-on-chronic liver failure (ACLF) rats; To further reveal the possible mechanism ofJieduan Niwan Formula.MethodsSPF Wistar rats were randomly divided into normal group, model group andJieduan Niwan group. 13-week porcine serum injection followed withD-galactosamine and lipopolysaccharide joint acute attack was used to established ACLF model in all rats except for normal group. Rats inJieduan Niwangroup were orally givenJieduan NiwanFormula for 3 days before acute attack. Rats were sacrificed respectively at 4, 8 and 12 h after models were established. The expressions of Bid, Cytochrome C (Cyt C) and hepatocyte ultrastructure were detected by Western blot method, immunohistochemical analysis and transmission electron microscope, respectively.Results Compared with normal group, the levels of Cyt C and Bid in model group increased at 4 h and peaked at 8 h but decreased at 12 h (P<0.05); however, the levels of Cyt C and Bid inJieduan Niwan group were lower than those in model group at 4 and 8 h but higher at 12 h (P<0.05). The ultrastructures were significantly damaged in model rats, which severity was escalated through time; inJieduan Niwan group, the degree of injury decreased at each timing.ConclusionJieduan NiwanFormula can efficiently alleviate the hepatocyte damage in ACLF rats, and the mechanism might be involved in the inhibition of the mitochondrial apoptotic pathway.

Objective Respectively applying the treatment of biventricular pacing and right ventricular septal pacing in atrioventricular block,to compare the heart function influence of two kinds of pacing mode on pacemaker dependent patients, to provide evidence for the physiological pacing mode selection?Methods Enrolled 20 patients from January 2012 to March 2013 who should be placed in pacemakers, their primary disease was the second degree,high or third degree atrioventricular block,giving them three chamber pacemaker ( right atrial + biventricular ) each?Randomly divided into right ventricular septum pacing group ( group A, n=10) and biventricular pacing group( group B,n=10)?Twelve months later,each group crossed into the each other group and continued following?up for 12 months?After 24 months to obtain all the data to do the statistical analysis,including patients'6 min walking distance(6MWD),the Minnesota Heart Failure Quality of life score (MLHFQ),plasma N?terminal pro brain natriuretic peptide precursor(NT?proBNP),left ventricular ejection ejection fraction(LVEF),left ventricular diastolic end diastolic diameter(LVEDD),left ventricular contraction end diastolic diameter(LVESD),left ventricular twelve segmental 14W time standard deviation(Ts?12SD),left ventricular twelve segmental 14W time maximum delay(Ts?dif),the paced QRS qrsd?Results Compared with group B,the 6MWD and LVEF of 12,24 months after treatment of group A were significantly increased( ( 242?58 ±37?56) m vs?(347?42±36?59) m vs?(340?67±24?99) m;(39?97±5?84)% vs?(57?92±10?01)% vs?(60?50±10?06)%;P<0?05),QRSd and NT?proBN were significantly decreased((139?25±10?43) ms vs?(114?25±10?07) ms vs?(110?83±11?08) ms) ms;( 2 857?84±236?48) ng/L vs?( 2 144?26±301?43) ng/L vs? (2 025?91±307?42) ng/L;P<0?05)?Compared with before treatment,at 12 and 24 months after treatment,6MWD in group B was significantly increased(228?17+38?06) m,(329?33+46?28) m,(350?67+35?43) m, LVEF was significantly increased ( ( 40?25+11?24 )% vs? ( 59?50+9?14 )% vs? ( 60?17+10?29)%),QRSd significantly narrowed((142?42+10?66) ms vs?(118?58+994) ms vs?(116?25+10?59) ms) and NT proBNP levels significantly reduced((2 848?25+318?65) ng/L vs?(2 144?26+301?43) ng/L vs?( 2 025?91+30?742) ng/L) were,the difference had statistical significance ( P<0?05)?There was no significant difference on the different time between the groups ( P=0?05)?Comparisons between A and B group at the same treatment time,these indexes of detections were no statistical significance(P>0?05)?Conclusion Compared with the right septal pacing,biventricular pacing is of no significant advantages on the effect of cardiac function for patients with pacemaker dependent.

Objective To study the stability of anticoagulant peptide Hirulog-S and its lyophilized product, and to provide data on the storage conditions and clinical applications.Methods RP-HPLC was used to determine the content and the related substances of Hirulog-S and its lyophilized powder with influence factor test, accelerated test and long-term storage test.Results Light, temperature and humidity had no significant effect on the stability of Hirulog-S and its lyophilized powder in the influence factor test.The content and related substances of Hirulog-S and its lyophilized powder did not significantly change in the accelerated test ( 40℃, RH75%) and 24-month long-term storage test at room temperature and 4℃.Conclusion Hirulog-S and its lyophilized product are very stable, even after being stored at room temperature for two years.