OBJECTIVE:To improve the teaching quality of pharmaceutical standard training. METHODS: By introducing the overview of adopting elicitation teaching in the pharmaceutical standard training in Beijing,the training effect was summarized and role and significance were studied. RESULTS:Supervisors applied five teaching modules in the elicitation teaching in the pharmaceu-tical standard training which are concluded“pharmaceutical education and rational use of medicine”“comments of prescription and rational use of medicine”“prescription transfers and rational use of medicine”“medicine counseling and rational use of medicine”“pharmaceutically communication and rational use of drugs medicine”,the result showed that participants had mastered more than 90%of what they had learned;with 92.5%of participants felt elicitation teaching could help improve students’interests in pharma-cy,and strengthen responsibility;80.0% of the participants believed that part of the modules strengthen the training of clinical think-ing;the training of satisfaction to 100%. CONCLUSIONS: The elicitation teaching mode can not only improve the teaching quality in pharmaceutical standard training,but also strengthen communication and feedback between teachers and students,students and students to make students more deeply understand and master knowledge and improve teaching quality.

Objective To investigate the clinical feature, treatment and prognosis of severe hand-footmouth disease without typical skin lesions. Methods Clinical data from 24 patients with severe hand-footmouth disease without typical skin lesions collected from January 2010 to May 2010 were retrospectively analyzed.Results There were 17 males and 7 females among the 24 patients. Of them, 3 patients with positive EV71 showed no skin lesions at the first visit, 2 presented with only herpes of mouth, 3 with only skin rashes in gluteal regions, 5 with only skin rashes on the palms, 2 with only skin rashes of the knees, 9 with skin rashes in hands, feet and gluteal regions simultaneously. The skin rashes were sparse with an atypical distribution.Fever occurred in all the 24 patients and lasted 6 - 7 days. Nervous system was involved in all the patients,and pulmonary hemorrhage occurred in 4 patients. RT-PCR and real-time PCR showed that the pharyngeal swab was positive for EV71 in 13 patients, for CoxA16 in 2, for other enterovirus in 3, and feces samples were positive for EV71 in 9 patients, for CoxA16 in 1, for other enterovirus in 2. Five patients were positive for EV71 in both pharyngeal swab and feces samples. Four patients died of multiple organ failure, the other 20 patients were cured and discharged from hospital. Conclusions The major pathogen causing severe hand-footmouth disease is EV71 in the 24 patients; children under 2 years are liable to this disease; high fever is common with various rashes. Early recognition of atypical skin lesions and timely management are most important for the control of severe hand-foot-mouth disease and improvement in overall survival of patients with this entity.

Objective To investigate the contents of IL-1β and IL-1ra in cerebrospinal fluid of children with Japanese encephalitis, and their clinical significance. Methods Enzyme linked immunosorbent assay (ELISA) was employed to detect IL-lβ and IL-lra contents in 50 children with Japanese encephalitis and 20 children without nervous system disease (controls). Results IL-1β contents in climax stage, convalescent stage and the controls were (49. 43±14. 59) , (24. 73±14. 50) and (8. 98± 1.26)μg/L (F = 79.88, P<0.01); IL-lra contents in climax stage, convalescent stage and the controls were (177. 39±60. 19), (78. 24±44. 63) and (21. 09±3. 10) μg/L (F = 91. 53, P <0. 01). There were significant differences on IL-lβ and IL-lra contents among children with mild, moderate and severe encephalitis (climax: F = 82.36 and 66.50, P<0.01; convalescence; F = 55. 17 and 79.50, P<0.01). IL-1β content was positively correlated with IL-lra in both climax and convalescent stages (climax; r = 0. 815, P < 0.01; convalescent; r= 0.728, P < 0.01). Conclusions IL-lβ and IL-lra contents in cerebrospinal fluid are significantly increased in children with Japanese encephalitis in climax stage, which are closely correlated with the disease severity. The two indicators may participate in the pathological process of brain damages with Japaness encephalitis.

Objective:To explore the quality of life and its influencing factors in patients with hyperthyroidism.Methods:From June 2017 to June 2019, patients with hyperthyroidism who were first diagnosed in the First Affiliated Hospital of Hainan Medical College were selected as the research objects, and the quality of life of the patients was assessed by the Chinese version of World Health Organization Quality of Life Brief Scale (WHOQOL-BREF). Hamilton Anxiety Scale (HAMA) and Hamilton Depression Scale (HAMD) were used to evaluate the psychological status of the patients. Univariate analysis was used to compare the differences of general information (age, gender, course of disease, marital status, education level, occupation, monthly family income, family relationship, thyroid ophthalmopathy, laboratory examination, thyroid crisis, anxiety level, and depression level) among patients with different quality of life. Multivariate logistic regression analysis was used to analyze the related factors affecting the quality of life of patients with hyperthyroidism.Results:There were 120 patients, aged (45.42 ± 6.38) years old, including 42 males and 78 females. The WHOQOL-BREF score was (76.58 ± 10.42) points, ranging from 25 to 106 points. Among them, 49 cases (40.83%) had high scores (≥80 points) and 71 cases (59.17%) had low scores (<80 points). Univariate analysis showed that there were differences in the quality of life among different marital status, education level, monthly family income, family relationship, thyroid ophthalmopathy, FT 3, FT 4, anxiety level and depression level ( P < 0.05). Multivariate analysis showed that thyroid ophthalmopathy, divorce or widowhood, monthly family income < 2 000 yuan, family disharmony, depression or anxiety, abnormal FT 4 and FT 3 were the risk factors affecting the quality of life of patients with hyperthyroidism ( P < 0.05). Conclusion:The overall quality of life of patients with hyperthyroidism is low, especially those with thyroid ophthalmopathy, divorce or widowhood, monthly family income < 2 000 yuan, family disharmony, depression, anxiety, FT 4 or FT 3 abnormalities.

Objective To construct the engineering bacteria expressing the recombinant human Kunitz protease inhibitor domain of amyloid protein precursor variant (rhKD/APPvar)in Pichia pastoris,and to establish the methods suitable for large-scale fermentation and purification of rhKD/APPvar.Methods The rhKD/APPvar expression vector was constructed based on the rhKD/APPvar-pPICZαexpression vector. Two restriction enzyme loci (ApaⅠ and SacⅡ)were added to two flanks of KD/APP and human KD/APP activity center RAM was replaced by the active site of BPTI KAR.After the rhKD/APPvar-pPICZαexpression vector was transformed into Pichiapastoris,optimized expression and purification of rhKD/APPvar was performed.The rhKD/APPvar was purified with cation exchange chromatography and desalting.Results The results of digestion identification and DNA sequencing analysis demonstrated that the recombinant plasmid rhKD/APPvar-pPICZα was successfully constructed and transfected into pastoris X-33. The SDS-PAGE analysis results indicated that rhKD/APPvar expressed after the induction of methanol and the relative molecular weight was 6 700.After a series of experiments the optimal expression conditions of rhKD/APPvar were obtained as follows:the optimal pH was 6.0 and the optimal induction time point was about the 5 th day for the strain.After purified the purity of rhKD/APPvar was about 95%.Conclusion KD/APPvar-pPICZ is successfully constructed;after expression in Pichia pastoris and purification,the rhKD/APPvar protein is obtained.

AIM: To observe the effect of osthole on tricalcium phosphate (TCP) particles-induced calvarial osteolysis in vivo.METHODS:Male ICR mice were randomly divided into sham group , TCP group and osthole group .A mouse calvarial model of osteolysis was established by TCP particles .On the second postoperative day , osthole (20 mg/kg) was locally injected into the calvarium under the periosteum 3 times a week.Two weeks after osthole treatment , blood and calvaria were collected to determine the level of bone turnover markers such as alkaline phosphatase ( ALP) , osteocalcin and tartrate-resistant acid phosphatase ( TRACP) .The periosteum was performed to examine the release of tumor necrosis factor-α(TNF-α), interleukin-6 (IL-6) and IL-1βby ELISA.The calvaria was obtained for histological and molecular analyses.RESULTS:Data from HE and TRACP staining revealed that osthole prevented TCP particles-induced obvious increase in osteoclastogenesis and resorption area in the metaphysis of mouse calvaria .Osthole treatment increased ALP ac-tivity and osteocalcin level , and dncreased the activity of TRACP in the mouse serum compared with TCP group .Further-more, TCP particles-induced the releases of TNF-α, IL-6 and IL-1βwere significantly suppressed by osthole treatment .In addition, Western blot demonstrated that endoplasmic reticulum ( ER) stress markers such as glucose-regulated protein 78 (GRP78) and CAAT/enhancer binding protein homologous protein (CHOP) were significantly up-regulated in TCP parti-cles-implanted calvarial mice , indicating that TCP particles triggered an ER stress response in the mouse calvarial osteolysis model , which obviously attenuated by osthole .CONCLUSION:Osthole inhibits TCP particles-induced calvarial osteolysis in mice, which is mediated by inhibition of ER stress signaling pathway .

Objective:To investigate the distribution of pathogen species isolated from cerebrospinal fluid culture (CSF) in children and analyze the antibiotic-resistance of the main isolates in vitro, which provides reference for interpreting the pathogens and choosing antibiotics in empiric therapy for pediatric patients. Methods:The results of cerebrospinal fluid culture were collected by checking laboratory information system of the Children′s Hospital of Zhejiang University and the clinical characteristics of these children were analyzed retrospectively by checking electronic medical record system.Results:A total of 1 312 isolates were detected, including 1 294 isolates of bacteria and 18 isolates of fungi. A total of 497 (37.9%) isolates were pathogenic microorganisms, of which 288 (57.9%) isolates were gram-positive, 200 (40.3%) isolates were gram-negative, and 9 (1.8%) isolates were fungi. The top 5 pathogens were Escherichia coli (102 isolates, 20.5%), Streptococcus pneumoniae (64 isolates, 12.9%), Streptococcus agalactiae (52 isolates, 10.5%), Enterococcus faecium (33 isolates, 6.6%) and Staphylococcus aureus (28 isolates, 5.6%). Most of the Streptococcus pneumoniae strains were isolated from children more than 1 year old (76.6%, 49/64), while the other top 4 bacteria were mainly isolated from infants less than 1 year old, with the rate of 95.1%(97/102) for Escherichia coli, 98.1%(51/52) for Streptococcus agalactiae, 81.8%(27/33) for Enterococcus faecium and 71.4% (20/28) for Staphylococcus aureus. A total of 815 (62.1%) isolates were considered to be contaminated pathogens according to the analysis on clinical manifestations and other laboratory findings in CSF, and coagulase-negative Staphylococcus (680 isolates), Micrococcus (50 isolates), Corynebacterium (28 isolates) and Enterococcus faecium (23 isolates), which accounted for 41.1% (23/56) of the total detected Enterococcus faecium, were the top 4 contaminated bacteria. During the study period, the isolation rate of the pathogenic microorganisms increased year by year (χ2=34.84, P<0.001), while the isolation rate of the contaminated pathogens, which detected mainly in summer and autumn, decreased year by year (χ2=13.26, P<0.001). Conclusions:The predominant bacteria causing pediatric purulent meningitis were Escherichia coli, Streptococcus pneumoniae, Streptococcus agalactiae, Enterococcus faecium and Staphylococcus aureus. Coagulase-negative Staphylococcus, Micrococcus, Corynebacterium and Enterococcus faecium were common contaminated bacteria in CSF culture, therefore clinicians should interpret the results of CSF culture cautiously according to the bacterial species and clinical manifestations.

Objective To investigate the clinical and laboratory features of severe cases of hand,foot and mouth disease (HFMD) in Wenzhou, Zhejiang Province. Methods Clinical data of 107 children with HFMD, including 97 severe and 10 critical cases treated in Children' s Hospital Affiliated to Wenzhou Medical College during January and May 2010 were retrospectively analyzed. One hundred and fifty children with mild HFMD were also selected as the controls. Clinical features and laboratory results were compared between the two groups. Results Fever, rash and infection in central nervous system were observed in all patients with severe HFMD, and symptoms on respiratory, digestive and cardiovascular systems were more serious than those of mild HFMD cases. White blood cell counts (WBC) were higher in severe group than those in controls (t = 12.72, P ＜0.01). Hyperglycemia (9. 2 mmol/L) and abnormal troponin (0. 3 -9. 0 ng/mL) were presented in all the critical patients. Cerebrospinal fluid WBC counts were raised in 97 severe HFMD patients (98.5 × 106/L for average) with predominance of lymphocytes. Among 107 severe patients, EV71 was positive in 70, including all 10 critical cases. Conclusion Involvement of nervous,respiratory and digestive symptoms is common in severe cases of HFMD, and EV71 is the predominant pathogen.

Objective To investigate Smac/DIABLO and cytochrome c(cyt-c)mRNA levels in liver tissue of rats with acute hepatic failure treated by microencapsulated hepatocyte.Methods Acute hepatic failure were induced by intraperitoneal injection of D-galactosamine in rats.and the rats were treated with microencapsulated hepatocytes,free hepatocytes and physical saline(contr01),respectively.Smac/DIABLO and cyt-c mRNA in liver tissue was detected by RT-PCR and the mRNA expression levels among three groups were compared.Results Smac/DIABLO and cyt-c mRNA levels in liver tissues of rats with acute hepatic failure were higher than those of normal rata(F=4.345,14.821,47.565,42.178 and 62.961,P<0.05).The peak values of Smac/DIABLO and cyt-C mRNA expressions in free hepatocytes and control groups were at 48 h.while that in microencapsulated hepatoeytes group was at 24 h.Conclusion Smac/DIABLO and cyt-c mRNA expression is an indicator of apoptosis of hepatocytes.

0.05).Conclusion In-vitro-cultured CB has good effects in the treatment of apoplexy.Neither in-vitro-cultrued CB nor natural CB for apoplexy has obvious adverse reaction.