OBJECTIVE To summarize the current status of position training for clinical pharmacists in China and provide references for the continuous optimization of such training programs. METHODS SinoMed， CNKI，VIP and Wanfang Data were electronically searched to collect position training of clinical pharmacists studies from the inception until November 5th 2024. After data extraction and quality evaluation， descriptive analysis was performed on the results of the included studies. RESULTS & A total of 68 pieces of relevant literature were included in the study. Among them， 50 studies reported on training content， 49 involved the allocation of teaching resources in the bases， 48 addressed training methods， and 39 focused on training evaluation； only 2 studies mentioned faculty development. There were notable variations in the clinical pharmacist training programs across different bases， particularly in the allocation of teaching resources， such as the composition of the teaching team and the utilization of auxiliary teaching tools. Additionally， differences existed in training approaches， such as those employing a single method versus a blended approach. Conversely， the core training content of each base generally revolved around clinical pharmacy practice， demonstrating a degree of consistency. Moreover， the overall emphasis on teacher training and assessment tended to be obviously insufficient. Each base can focus on enhancing the competence of clinical pharmacists by allocating teaching resources， selecting training methods， improving training content， and using evaluation tools， to further enhance the quality of clinical pharmacist training.

Objective:A mouse model of pancreatitis induced by coxsackievirus B3 (CVB3) was established. The pathological change of pancreas and the infiltration of Th17/Treg cells were observed.Methods:The BALB/c mice were inoculated intraperitoneally with CVB3 to induce acute viral pancreatitis model. Then the pathological changes of pancreas were observed by HE staining; the viral RNA load and relative expression of cytokines (IFN-γ, IL-6 and IL-17) mRNA were detected by q-PCR; the proportion of infiltrated CD45 + CD3 + T cells, CD4 + and CD8 + T cells, Th17 and Treg cells in the pancreas was determined by flow cytometry. Results:Three days after CVB3 infection, the viral RNA load in pancreas was the highest (0.96±0.18) and gradually decreased with prolongation of infection. Compared with the 3 dpi group, the viral RNA load in pancreas was decreased (0.96±0.18 vs. 0.62±0.14) at 7 dpi, but there was no statistically significant difference. In addition, the infiltration of immune cell in pancreas increased significantly after 7dpi and the pathological score >2. The percent of infiltrated Th17 cells (1.05±0.21 vs. 22.13±5.79) and Treg cells (3.11±0.78 vs. 8.25±1.30) among CD4 + T cells significantly increased after infection (P<0.05), and the Th17/Treg also increased (P<0.01). Compared with the control group, the relative mRNA expression of IFN-γ (1.05±0.23 vs. 672.6±47.67), IL-6 (1.00±0.38 vs. 68.28±4.57), and IL-17 (1.01±0.11 vs. 54.15±7.94) in pancreas increased at 7 days after CVB3 infection ( P<0.01). Conclusions:The infiltration of Th17/Treg cells and the expression of related cytokines related cytokines IL-6 and IL-17 mRNA were upregulated in pancreas, which promoted the process of CVB3-induced pancreatitis.

Objective:To analyze the association of respiratory syncytial virus infection with meteorological factors and to predict and explain the trends.Methods:Data of cases with severe acute respiratory infections in hospitalized children in Xuzhou City were collected from 2015-2021. Respiratory syncytial virus (RSV) was detected by real-time fluorescence polymerase chain reaction. The result were statistically analyzed using SPSS 26.0 software, including constructing a negative binomial regression model to explore meteorological factors that impact RSV detection and a multivariate time series model to predict its epidemiological trend from 2020 to 2021.Results:A total of 1 663 samples of children with severe acute respiratory infections were collected from 2015to 2021, of which 218 (13.1%) were positive for RSV. Seasonal effects on RSV detection were evident: there was a 1-year cycle with a peak in winter (December-February) and a trough in summer (June-August). The negative binomial regression analysis showed that monthly mean temperature, monthly mean relative humidity, and monthly total sunshine hours may influenced RSV detection. The prediction result of the time series model with sunshine hours as the covariate showed that the prediction was better for 2020, and the actual values were close to the predicted values. The expected trends in 2021 were consistent, but the actual values were higher than predicted.Conclusions:Monthly mean temperature, monthly mean relative humidity, and monthly total sunshine hours may influence RSV detection in the Xuzhou region.A prediction model can be built using data from 2015-2019, where deviations in the predicted values for 2021, reflecting that disease prevalence is multifactorial correlated, suggest a possible rise in RSV prevalence in the future.

Objective:To establish a real-time fluorescent quantitative PCR for the detection of torque teno virus types 7 (TTV7), 8 (TTV8) and 10 (TTV10) and analyze its performance in clinical sample detection.Methods:Specific primers were designed based on the gene sequences of TTV7, TTV8 and TTV10 in GenBank. Recombinant plasmids of pMD19-T-TTV7, pMD19-T-TTV8 and pMD19-T-TTV10 were constructed and used as positive standard control to establish a real-time fluorescent quantitative PCR based on FAM-Eclipse probe method. The specificity and sensitivity of the established method were evaluated. Moreover, it was validated in terms of clinical sample detection.Results:The standard curve equations of the real-time fluorescent quantitative PCR for detecting TTV7, TTV8 and TTV10 were y=-0.340 2 x+ 114.780 0 ( R2=0.998 8), y=-0.351 1 x+ 114.940 0 ( R2=0.995 3) and y=-0.348 9 x+ 115.020 0 ( R2=0.991 7), respectively, and there was no cross-reaction with other viruses. The detection sensitivity of the established method for TTV7, TTV8 and TTV10 were 108 copies/μl, 84 copies/μl and 98 copies/μl, and the positive detection rates in clinical pediatric serum samples were 10.9%, 2.1% and 4.3%, respectively. Conclusions:The established real-time fluorescent quantitative PCR for detection of TTV7, TTV8 and TTV10 was featured by strong specificity and high sensitivity, which could be used for rapid TTV detection in clinical serum samples.

Objective:To investigate the incidence and clinical characteristics of Kawasaki disease complicated with arthritis, and explore the relationship with coronary artery disease.Methods:Patients diagnosed with Kawasaki disease at Xiamen Children′s Hospital from January 2015 to June 2020 were included in this study.They were divided into the arthritis group( n=53) and the non-arthritis group( n=401), depending on whether complicated with arthritis.Demographic, clinical symptoms, and laboratory results were retrospectively analyzed. Results:A total of 454 children were included in this study with 53 cases acomplicated with arthritis.There were 32 male cases and 21 female cases.The average age of arthritis group was(5.89±1.35) y, which was older than non-arthritis group[(4.28±1.25) y, P=0.026]. Among the 53 cases of arthritis group, 36 cases (67.92%)of small jiont arthritis, 14 cases(26.41%)of coxitis, ten cases(18.87%)of carpitis, eight cases(15.09%)of gonitis, four cases(7.55%)of anconitis, and three cases(5.66%) of ankle arthritis were involved.There was a statistic difference in the prevalence of intravenous immunogloblin(IVIG)resistant between arthritis group and non-arthritis group(14 cases, 26.14% vs.43 cases, 10.72%, P=0.002). The inflammatory markers(CRP, TNF-α, IL-6) of the arthritis group were significantly higher than those in the non-arthritis group, and the differences were statistically significant( P<0.05, respectively). The incidence of coronary artery disease in the arthritis group(60.38%, 32/53) was higher than that in the non-arthritis group(52.37%, 210/401), but the difference was not statistically significant( P>0.05). Conclusion:Kawasaki disease with arthritis in children is self-limited, with no sequelae.Patients in the arthritis group have a higher rate of IVIG resistance and higher levels of inflammatory markers, but no significant difference in the incidence of coronary artery disease compared with those without arthritis.

Objective:To construct the dual-luciferase reporter vector for identification of internal ribosome entry site (IRES).Methods:The hairpin structure was inserted between Renilla luciferase (R-Luc) and Firefly luciferase (F-Luc) genes based on psiCHECK-2 to form plasmid psiCHECK-IRES. IRES of Encephalomyocarditis virus (EMCV) was inserted between the hairpin structure and F-Luc genes of psiCHECK-IRES to form vector psiCHECK-IRES-EMCV. After psiCHECK-IRES-EMCV or psiCHECK-IRES was transfected into BHK-21 cells respectively, expressions of F-Luc and R-Luc were detected by RT-qPCR. Then Luciferase activity of transfected cells was detected with the dual-luciferase reporter assay system at 24 h post-transfection.Results:The hairpin structure was successfully inserted into psiCHECK-2 to form psiCHECK-IRES by sequencing. RT-qPCR result showed that there were the approximate expressing levels of mRNA between F-Luc and R-Luc. The result indicated that no aberrant monocistronic transcripts, which caused false positive F-Luc readings, were produced. Then IRES of EMCV was introduced into psiCHECK-IRES to form psiCHECK-IRES-EMCV. The F-Luc/R-Luc ratio in psiCHECK-IRES-EMCV-transfected cells was 53.35 times that of psiCHECK-IRES-transfected cells. The result confirmed that IRES of EMCV initiated effectively the translation of F-Luc.Conclusions:Dual-luciferase reporter vector psiCHECK-IRES was successfully constructed, which could be used to validate viruses and eukaryotic genes, the translation thereof was IRES-dependent.

Objective:To study the feasibility of using bedside ultrasound in evaluating gastric residual volume in critical ill patients with enteral nutrition support.Methods:From May 2019 to August 2019, 60 patients were selected to receive enteral nutrition via gastric tube in ICU of Union Hospital, Tongji Medical College, Huazhong University of Science and Technology. Patients were divided into the experimental group and the control group according to the odd and even number of beds, 30 patients in the experimental group with odd number of beds and 30 patients in the control group with even number of beds. Gastric residual volume was evaluated at 0, 4, 8, 12, 16, 20, 24 h of enteral nutrition. In the experimental group, the gastric residual volume was evaluated by bedside ultrasound and syringe suction at each time point. In the control group, only bedside ultrasound was used to evaluate gastric residual volume. The results of operation time, monitoring results at different time points, diarrhea and the utilization rate of gastrointestinal motility drugs target feeding time, vomiting, were compared between the two groups.Results:There was no statistical difference between the gastric residual amount monitored by ultrasound and the gastric residual amount monitored by suction ( P>0.05). The operating time of bedside ultrasound monitoring was (62.40 ± 4.00) s, the operating time of suction monitoring was (78.39 ± 12.15) s, and the operating time of bedside ultrasound monitoring was less than that of suction ( t value was 6.633, P<0.01). There was no significant difference in the rate of vomiting, diarrhea and gastrointestinal motility drugs between the two groups( P>0.05). The time to reach the target feeding amount in the control group was (3.04 ± 0.31) d, and the time to reach the target feeding amount in the experimental group was (4.19 ± 0.33) d. The time to reach the target feeding amount in the control group was less than that in the experimental group ( t value was 13.42, P<0.01). Conclusions:Bedside ultrasound can be used to evaluate the residual gastric volume of enteral nutrition support patients, guide the implementation of enteral nutrition, shorten the operation time, reduce the workload of nurses, and avoid the contamination of enteral nutrition preparation.

Objective: To establish a real-time quantitative PCR detection system for Torque teno virus (TTV) and verify the sensitivity and specificity of the detection system. Methods: Primers and FAM-Eclipse probes were designed based on the TTV6 gene sequence registered in GenBank, and were to establish a real-time fluorescent quantitative PCR detecting way based on the FAM-Eclipse probe, the standard curve was constructed and sensitivity and specificity were analyzed. Results: A quantitative PCR method for the specific detection of TTV6 were established that the standard curve equation was y=-3.0921x + 28.36, and the amplification efficiency and R2 were 99.6% and 1.000, respectively. The sensitivity of TTV6 was 1.0×10 copies/μl, and there was no cross-reactivity with other viruses. There was 1 case positive for TTV6 out of 56 throat swab samples from the patients with clinical respiratory infection. Conclusions The real-time fluorescent quantitative PCR for detecting TTV6 established by FAM-Eclipse probe had the advantages of high sensitivity and specificity. It provides an effective way for detection and quantification of viral content of TTV6 in clinical specimens.

Objective: To study the intracellular location and autophagosome production of rhinovirus 16 2B protein using miniSOG labeling technique. Methods: 2B was fused with miniSOG and flag tags to construct pcDNA3.1-2B-miniSOG-flag plasmid, which was used to transfect HEK293 cells, LC3 protein was detected by western blot. The transfected cells were fixed, stained with DAB through the photooxidation activity of miniSOG, and used to prepare ultrathin sections. Localization of 2B-miniSOG protein in cells and ultrastructural changes of cells were observed under electron microscope. Results: 2B-miniSOG protein glows green under a fluorescence microscopy. Green flourescence coold be observed in the cells expressing 2B-miniSOG protein.LC-II protein increased in the cells transfected with pcDNA3.1-2B-miniSOG-flag. Under electron microscopy it was observed that 2B-miniSOG protein was located in the mitochondria, and a large number of vesicular structures appeared in the cytoplasm. Both autophagosomes and autophagic lysosomes can be observed. Conclusions Non-structural protein 2B of HRV16 can induce autophagy.

Objective: To investigate the effect of thapsigargin (TG) which can induce endoplasmic reticulum stress (ERS) on the replication of coxsackievirus B 3 (CV-B3). Methods: After 10 MOI CV-B3 infected HeLa cells were exposed 0.25 μmol/L TG for 3 h, 6 h and 9 h, virus RNA of HeLa cells were extracted and viral replication was evaluated by real time PCR. After 0.25 μmol/L、0.08 μmol/L and 0.025 μmol/L TG exposed, the plaque of CV-B3 was used to confirm further replication of CV-B3. To verify TG induced ERS through three signal pathway, one of among PERK, ATF6 and IRE1 inhibitors GSK2656157, AEBSF and STF-083010, and 0.25 μmol/L TG were used in HeLa cells infected with 10 MOI CV-B3, replication of CV-B3 was evaluated by qRT-PCR. Results: The stimulation of TG did not induce increase of virus replication after post-infection 3 h. However, TG induced replication of virus to increase 2.5 times after post-infection 6 h and 158.6 times after post-infection 9 h. And, the area of viral plaque was significantly increased. ATF6 inhibitors AEBSF significantly inhibited promotion of virus replication from TG. Conclusions TG can promote the replication of CV-B3 through ATF6 signal pathway.