Objective:To determine the alteration of miRNA profile of human tonsillar epithelial cells induced by enterovirus-A71 (EV-A71) infection.Methods:Human tonsillar epithelial cells UT-SCC-60B were infected with EV-A71 at multiplicities of infection (MOI) of 1 and total RNA was extracted using Trizol reagent. Small RNA library was constructed and high-throughput sequencing was performed using Illumina NextSeq 500. Differential significantly expressed known and novel miRNAs and putative targets were selected after the processing of raw data. Gene ontology (GO), kyoto encyclopedia of genes and genomes (KEGG) pathways were analyzed through online database. Kinds of miRNA could target EV-A71 genome was determined through psRNATarget. Validations of random selected miRNAs were done through real-time RT-PCR.Results:A total of 61 known significantly expressed miRNAs (21 miRNAs were down-regulated and 40 miRNAs were up-regulated) and 559 novel significantly expressed miRNAs were identified through high-throughput sequencing. Novel significantly expressed miRNA had typical "hairpin structure" of pre-miRNA. Fold changes of hsa-miR-517b-3p and hsa-miR-199a-5p which was determined by real-time RT-PCR had similar change trends with high-throughput sequencing. Putative targets of significantly expressed miRNA were referred to different biological processes and signaling pathways. A total of 24 significant miRNAs (5 known significantly expressed miRNAs and 19 novel significantly expressed miRNAs) had "seed sequence" in EV-A71 genome.Conclusions:Expression of miRNA profile in UT-SCC-60B was significantly changed by EV-A71 infection and the identified significantly expressed miRNAs potential target EV-A71 genome to regulate EV-A71 replication.

Objective:To investigate the degree of psychological pain in patients with gastrointestinal cancer after multiple chemotherapy, and analyze the influencing factors of psychological pain.Methods:Totally 130 patients with gastrointestinal tumors admitted from December 2017 to December 2019 admitted to the People′s Hospital of Jiangsu Province were included, and all patients received multiple chemotherapy. Using the psychological pain screening tools to evaluate the degree of psychological pain of patients. The data of patients were collected and statistically analyzed by single factor and binary logistics regression model.Results:Among the 130 patients, 32 cases(24.62%) had no psychological pain and 98 cases (75.38%) had psychological pain. There were differences in the age, pathological stage, distant metastasis, monthly family income, accompanying staff, hospitalization time, and personality type of the mentally distressed and psychologically distressed ( χ2 values were 4.598-12.404, P<0.05). Binary logistics regression model analysis suggested that age, pathological stage, distant metastasis, family monthly income, accompany status, length of hospital stay, and personality type were the influencing factors of patients' psychological pain ( P<0.05). Conclusions:Psychological pain of patients with gastrointestinal tumors after multiple chemotherapy is more common, and its occurrence is mainly related to the patient′s age, pathological stage, distant metastasis, family monthly income, accompanying staff, hospitalization time and personality type.

Objective: To investigate the role of des-gamma-carboxy prothrombin (DCP) in assessment of liver function and prognosis of patients with liver cirrhosis. Methods: From January 2013 to August 2016, a total of 137 patients with liver cirrhosis in Shanghai Changzheng Hospital were enrolled. The serum DCP level was measured, the clinical data was collected and the complication and survival situation was followed up. The 137 patients were divided into DCP negative group (DCP≤40 mAU/mL, 118 cases) and DCP positive group (DCP>40 mAU/mL, 19 cases). Forty-five patients with compensated liver cirrhosis were divided into high-level DCP group (DCP>16.5 mAU/mL, 32 cases) and low-level DCP group (DCP≤16.5 mAU/mL, 13 cases). Chi square test was used to analyze the difference in the positive rate of DCP in patients with different Child-Pugh classification. Spearman correlation test was performed to analyze the correlation between DCP and model for end-stage liver disease (MELD) scores. Kaplan-Meier survival curve was used to analyze the correlation between DCP and liver disease related mortality. Results: Compared to that of DCP negative group, albumin level of patients in DCP positive group decreased (35 g/L, 20 to 57 g/L vs. 29 g/L, 17 to 42 g/L), however, total bilirubin (TBil), prothrombin time (PT), and international normalized ratio all increased (12.9 mg/L, 1.80 to 83.0 mg/L vs.22.2 mg/L, 6.4 to 169.0 mg/L; 15.5 s, 11.7 to 35.7 s vs.17.5 s, 13.9 to 33.4 s; 1.24, 0.96 to 3.72 vs.1.44, 1.09 to 3.22), and the differences were statistically significant (Z=-2.785, -2.891, -2.945 and -2.879, all P<0.01). The DCP positive (DCP>40 mAU/mL) rates of Child-Pugh A, B and C patients were 1.8% (1/55), 21.2% (11/52) and 23.3% (7/30), respectively, and the difference was statistically significant (χ²=11.246, P=0.003). The DCP levels of patients with Child-Pugh class B and C cirrhosis were significantly correlated with MELD scores (r=0.259, P=0.021). There were 16 and three patients with ascites and spontaneous bacterial peritonitis (SBP) of DCP positive group, and the incidence was higher than that of DCP negative group (55.1%, 65/118 and 1.7%, 2/118), and the differences were statistically significant (χ²=5.744 and 97.636, both P<0.05). Patients in high-level DCP group had a higher proportion of clinical decompensation than low-level DCP group (53.1% (17/32) and two cases), the difference was statistically significant (χ²=5.397, P=0.024). The overall survival rate of DCP positive group (nine survival cases) were lower than that of DCP negative group (87.9%, 87/99), and the difference was statistically significant (χ²=5.442, P=0.020). Conclusions Serum DCP level is closely related to liver function, ascites and SBP in patients with liver cirrhosis. Furthermore, it is associated with occurrence of decompensation in compensated patients and liver-related mortality in patients with liver cirrhosis. DCP may be a useful serum marker for evaluation of disease severity and prognosis in patients with liver cirrhosis in clinical practice.

Objective To investigate the effect of calcium on the stability of VWF-A2 domain. Methods The crystal structures of A2 (not containing calcium) and A2/Ca2+ (with calcium bound) were downloaded from protein data bank. For A2 domain, the conformational changes, unfolding pathway differences and the exposure degree variance of cleavage sites caused by calcium binding were observed and analyzed by steered Molecular Dynamics simulations under constant force. Results The unfolding pathway of A2 domain and exposure process of cleavage sites were force-dependent. Calcium binding did not affect the unfolding process of A2 in the early stage. As the conformational rearrangement of α3β4-loop reduced its localized dynamic properties, the movement among β1-β4-β5 strands was restrained, which suppressed its further unfolding to stay in the intermediate steady state and delayed the cleavage-site exposure. Conclusions Stretch force could induce β5 strand of A2 unfolding and the cleavage-site exposure, while calcium binding inhibited ADAMTS13 proteolysis efficiency through stabilizing A2 hydrophobic core and covering its cleavage sites. These results way help to understand how ADAMTS13 cleavages the VWF-A2 domain and regulates the hemostatic potential of VWF, and further provide useful guidance on the design of related anti-thrombus drugs.

Objective To explore the effect and significance of motor imagery therapy in the rehabilitation of stroke patients with hemiplegia. Methods A total of 60 stroke patients with hemiplegia were randomly assigned to the observation group(n=31) and the control group(n=29). Rehabilitation training and guidance of control group adopted interpretation of the demonstration method while observation group adopted motor imagery therapy. Movement function, daily life activities and satisfaction were compared between two groups after 4 weeks and 8 weeks. Results The differences in motor function, daily life activities and satisfaction were significant (P<0.05).Conclusions The use of motor imagery therapy in rehabilitation training and guidance of stroke patients with hemiplegia can improve the effect of health guidance, improve the motor function and daily life activities, promote the interaction and communication between nurses and patients, and improve the patients′satisfaction.

Objective To investigate the effect and safety of transcutaneous electrical stimulation at Zusanli acupoint for elimination of gastrointestinal poison in patients with oral organophosphorus pesticide poisoning. Methods A prospective study was conducted, including 62 patients with oral organophosphorus pesticide poisoning from September 2013 to February 2015 in the Department of Emergency of Xingtai People's Hospital of Hebei Province. The patients were divided into a observation group and a control group (each, 31 cases) in accord with the principle of simple random sampling. All the patients in two groups were given comprehensive treatment according to the diagnosis and treatment of the guide for organic phosphorus pesticide poisoning. In addition, the transcutaneous electrical stimulation at bilateral Zusanli acupoints was carried out in the observation group, once therapeutic time 30 minutes and every 8 hours once, and stopped until the discharge of melena. The incidence of vomiting after application of cathartics, the first stool time, the time of melena discharge, daily defecation frequency, the time of reaching atropinization, the total amount of atropine used, the time of cholinesterase (ChE) activity returning to its 1/2 normal activity, the length of stay in hospital, cure rate and mortality were observed in both groups. The changes in hemodynamics and pulse blood oxygen saturation (SpO2) were observed before and after transcutaneous electrical acupoint stimulation, and the occurrence of adverse reactions in the therapeutic course were observed in the observation group.Results After application of cathartics, the incidence of vomiting in control group was significantly higher than that in the observation group [32.2% (10/31) vs. 9.7% (3/31),P < 0.05]. In the observation group, the first stool time (hours: 9.3±3.6 vs. 11.6±5.2) and the time of melena discharge (hours: 11.3±5.3 vs. 14.5±6.8) were significantly shorter than those in the control group (both P < 0.05); while the frequency of bowel movements during catharsis was higher than that of the control group (times/d: 4.3±0.5 vs. 3.1±0.4,P < 0.01). In the observation group, the time reaching atropinization (hours: 66.3±22.8 vs. 84.6±24.2), the total amount of atropine used (mg: 66.3±22.8 vs. 84.6±24.2), and the time of ChE activity returning to its 1/2 normal range (days: 6.1±2.4 vs. 8.3±3.9) were significantly shorter than those in the control group (allP < 0.01). At the end of treatment, the average length of stay in hospital was shorter (days: 11.3±2.8 vs. 13.4±4.2,P < 0.05) and the cure rate was higher [96.8% (30/31) vs. 83.9% (26/31),P < 0.05] in the observation group than those in the control group; in observation group, the hemodynamics and SpO2 before and after acupoint electrical stimulation did not change significantly; in the course of treatment, no adverse reactions occurred.Conclusion The addition of transcutaneous electric stimulation at bilateral Zusanli acupoints in patients with oral organophosphorus pesticide poisoning has following advantages: lowering the incidence of vomiting during catharsis, enhancing the cathartic effect, promoting gastrointestinal poisoning discharge as soon as possible, reducing total atropine used during hospitalization, shortening the time reaching atropinization, shortening the duration of hospitalization, promoting the recovery of cholinesterase activity and elevating clinical therapeutic effects.

Objective There is little research on the relationship of gene polymorphism of CYP2C19 and clopidogrel response after percutaneous transluminal angioplasty and stenting ( PTAS) in patients with ischemic cerebrovascular disease.The study aimed to investigate the relationship between gene polymorphism and high on-treatment platelet reactivity ( HTPR ) after PTAS and 6 months of regular dual antiplatelet administration in patients. Methods A total of 145 Chinese patients treated with PTAS in our de-partment from January 2011 to March 2014 were enrolled in this study.According to the gene sequencing, patients were divided into wild-type group(CYP2C19*1/*1,69 cases) and mutation group(heterozygous mutation CYP2C19*1/*2 and homozygous mutation CYP2C19*2/*2, 76 cases).Patients received a 100mg/d aspirin and 75mg/d clopidogrel maintenance dose (MD) as dual anti-platelet therapy after PTAS.The clopidogrel inhibition effect was measured by thrombelastography ( TEG) system 6 months after PTAS. Routine cerebral artery digital subtraction angiography was applied to evaluate whether there was restenosis in stent and logistic regres-sion analysis was used to analyze the influential factors of HTPR after PTAS and clopidogrel adminstration. Results After 6 months'regular administration of clopidogrel after PTAS, the platelet adenosine diphosphate ( ADP ) receptor inhibition rates in wild-type group, heterozygous mutation and homozygous mutation group were respectively (58.43 ±21.98)%, (47.80 ±22.93)%, (37.53 ± 21.84)%.The platelet ADP receptor inhibition rate was significantly decreased compared with wild-type group(P=0.001).Carriers of at least one CYP2C19 loss-of-function ( LOF) allele had a higher frequency of clopidogrel HTPR (35.5% vs17.4 % for patients with and without LOF alleles, respectively;P=0.014) .Using multivariate logistic regression analysis, the carriage of CYP2C19 LOF alleles was an independent predictor of the post-procedure HTPR (OR=2.356, 95% CI:1.053-5.272, P=0.037).The rate of ISR was remarkably higher in patients with at least one CYP2C19*2 alleles compared with wild-type patients(11.8%vs 1.4%, P=0.019) . Conclusion In patients with ischemic cerebrovascular disease, the CYP2C19 LOF allele had significant impact on post-procedure clopidogrel HTPR and the prognosis of ISR after PTAS.

Objective:To provide the basic data for the further revision of mycoplasma test method described in Chinese Pharma-copoeia and some references for the operation standardization of drug mycoplasma test. Methods:Two incubation conditions,namely aerobic conditions and microaerophilic conditions,were compared with respect to the growth status of mycoplasma in liquid and solid media. Results:The growth of mycoplasma was obvious difference between the two incubation conditions,and the microaerophilic con-ditions were better than the aerobic conditions. Conclusion:The microaerophilic conditions can be used in the incubation of mycoplas-ma test,which should be defined and standardized in the future Chinese Pharmacopoeia.

Microbiological data deviations ( MDD) investigation differs from out of the standard ( OOS) investigation in chemical inspection items. Investigating Out-of-Specification( OOS) Test Results for Pharmaceutical Production issued by FDA specifically ex-cludes microbiological and other biological assays. Until recently, there is not a definitive guideline for MDD investigation in the world. Some institutions and experts have explored the practice and theory of MDD investigation. Referred to the existing practical experience and theoretical achievement and based on the provisions of Chinese GMP (2010 revision) and the requirements of Chinese Pharmaco-poeia Ⅱ(2010 edition) Appendix XIX Q, the article discussed the methods for MDD laboratory investigation.

Objective:To investigate the growth-promoting properties and applicability of the mycoplasma test culture media pre-scribed in Chinese pharmacopoeia, and provide reference for the standardization of the drug mycoplasma test method. Methods: Re-spectively using four quantitative detection methods including sensitivity, degree of color change, colony counts and colony diameter, and mycoplasma media widely used in the world as the reference media, the growth-promoting properties of 4 batches of mycoplasma broth and 4 batches of arginine mycoplasma broth from four domestic manufacturers were investigated. Results:The results of sensitivity assay and absorbance detection showed that all the media inoculated with below 100 CFU test microorganisms exhibited visible color change. Furthermore, the results of color change degree and colony diameter showed that there were significant differences among the media products from different manufacturers(P<0. 01). Conclusion:Mycoplasma broth and arginine mycoplasma broth both can sup-port the growth of below 100 CFU test microorganisms. Due to the difference in the growth-promoting properties among the media prod-ucts from different manufacturers, the drug mycoplasma test workers should use more sensitive methods to examine the applicability of the media.