OBJECTIVE To establish the index system of quality evaluation standard for pharmacist training in cough and wheeze pharmaceutical care （CWPC） outpatient department， and to provide a basis for the selection of CWPC pharmacist training teachers and the improvement of training plans. METHODS Based on Kirkpatrick model， using the Delphi method， a total of 15 experts from 13 tertiary hospitals in 10 provinces in China were consulted to establish the standard index system of the quality evaluation for CWPC pharmacists training. Analytic hierarchy process （AHP） was adopted to determine the weights of each indicator and quantify the index system according to the weights of indicators at all levels. RESULTS The coefficient of expert authority was 0.810， the judgment coefficient was 0.727， and the familiarity coefficient was 0.893. The Kendall coordination coefficient of each index was 0.308-0.687. The P values of χ2 test were all less than 0.05， which indicated that the degree of coordination of the experts was high. After two rounds of correspondence， Kirkpatrick model-based index system of quality evaluation standard for CWPC pharmacist training was determined. The index system included 4 first-level indexes （participants’ reaction layer， learning gain layer， behavior improvement layer， training outcome layer）， 12 second-level indexes （such as training needs， teaching methods， theoretical knowledge， practical skills， job abilities， patient benefits， etc.） and 44 third-level indexes （such as clear training objectives， core system of CWPC， special device operation and evaluation， professional knowledge related to the treatment of cough and wheeze patients， promoting the construction of CWPC， improving patient compliance）. CONCLUSIONS The constructed index system of quality evaluation standard for CWPC pharmacist training has a certain level of authority and scientificity， and provides a scientific theoretical basis for quality evaluation of CWPC pharmacist training.

Objective:To investigate the effects of osimertinib on proliferation, migration and invasion of procollagen-lysine 2-oxoglutarate 5-dioxygenase 2 (PLOD2) overexpressing HCC827 cells and explore the potential mechanism of PLOD2 induced osimertinib resistance.Methods:We transfected HCC827 cells with LV-vector and LV-over/PLOD2. The expression of PLOD2 was detected by quantitative real time polymerase chain reaction (qRT-PCR) and western blotting. The effects of osimertinib on the proliferation of HCC827-vector and HCC827-PLOD2 cells were evaluated by 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2H tetrazolium bromide (MTT) assay. The effects of osimertinib on the migration and invasion of HCC827-vector and HCC827-PLOD2 cells were determined by Transwell assays. The expressions of E-cadherin and vimentin in cells were detected by immunofluorescence (IF). The expressions of epithelial-mesenchymal transition (EMT), FAK-PI3K/AKT and MAPK signal pathway related proteins were detected by western blotting.Results:The MTT assay showed that HCC827-PLOD2 cells were hyposensitive to osimertinib. The 50% inhibitory concentration (IC 50) and resistance index of osimertinib for HCC827-PLOD2 cells was over 1 000 nmol/L and over 100, respectively. The result of wound healing assay showed that the migration distance of HCC827-PLOD2 was about (2.13±0.21) fold changes as that of HCC827-vector cells. The result of Transwell assay showed that the numbers of HCC827-PLOD2 passing through the matrix membrane were (212.78±10.43), significantly higher than (101.32±12.52) of HCC827-vector cells ( P<0.01). The result of IF showed that compared with HCC827-vector cells, the expression of E-cadherin was down-regulated while vimentin was up-regulated in HCC827-PLOD2 cells. Osimertinb downregulated E-cadherin and upregulated vimentin expression in HCC827-vector cells but had limited effect in HCC827-PLOD2 cells. The result of western blotting showed that PLOD2 significantly increased vimentin expression level while decreased E-cadherin expression level. Osimertinib inhibited the expression of p-EGFR, but did not affect the expressions of PLOD2, p-FAK, p-AKT, p-ERK, vimentin and E-cadherin in HCC827-PLOD2 cells. Conclusion:PLOD2 confers resistance to osimertinib in HCC827 cells by regulating EMT, FAK-PI3K/AKT and MAPK signal pathways.

Objective:To investigate the effects of osimertinib on proliferation, migration and invasion of procollagen-lysine 2-oxoglutarate 5-dioxygenase 2 (PLOD2) overexpressing HCC827 cells and explore the potential mechanism of PLOD2 induced osimertinib resistance.Methods:We transfected HCC827 cells with LV-vector and LV-over/PLOD2. The expression of PLOD2 was detected by quantitative real time polymerase chain reaction (qRT-PCR) and western blotting. The effects of osimertinib on the proliferation of HCC827-vector and HCC827-PLOD2 cells were evaluated by 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2H tetrazolium bromide (MTT) assay. The effects of osimertinib on the migration and invasion of HCC827-vector and HCC827-PLOD2 cells were determined by Transwell assays. The expressions of E-cadherin and vimentin in cells were detected by immunofluorescence (IF). The expressions of epithelial-mesenchymal transition (EMT), FAK-PI3K/AKT and MAPK signal pathway related proteins were detected by western blotting.Results:The MTT assay showed that HCC827-PLOD2 cells were hyposensitive to osimertinib. The 50% inhibitory concentration (IC 50) and resistance index of osimertinib for HCC827-PLOD2 cells was over 1 000 nmol/L and over 100, respectively. The result of wound healing assay showed that the migration distance of HCC827-PLOD2 was about (2.13±0.21) fold changes as that of HCC827-vector cells. The result of Transwell assay showed that the numbers of HCC827-PLOD2 passing through the matrix membrane were (212.78±10.43), significantly higher than (101.32±12.52) of HCC827-vector cells ( P<0.01). The result of IF showed that compared with HCC827-vector cells, the expression of E-cadherin was down-regulated while vimentin was up-regulated in HCC827-PLOD2 cells. Osimertinb downregulated E-cadherin and upregulated vimentin expression in HCC827-vector cells but had limited effect in HCC827-PLOD2 cells. The result of western blotting showed that PLOD2 significantly increased vimentin expression level while decreased E-cadherin expression level. Osimertinib inhibited the expression of p-EGFR, but did not affect the expressions of PLOD2, p-FAK, p-AKT, p-ERK, vimentin and E-cadherin in HCC827-PLOD2 cells. Conclusion:PLOD2 confers resistance to osimertinib in HCC827 cells by regulating EMT, FAK-PI3K/AKT and MAPK signal pathways.

Objective: To test the effect of metastasis associated in lung adenocarcinoma transcript 1 (MALAT1) and/or osimertinib on the proliferation and apoptosis of HCC827 cells, and explore the potential mechanism of MALAT1 induced resistance to osimertinib. Methods: We transfected HCC827 cells with LV-vector or LV-over/MALAT1. Stable transfected cells (HCC827/Vector, HCC827/MALAT1) were selected by adding puromycin. HCC827/MALAT1 cells were further transfected with the shRNA-negative control (NC) or shRNA-human epidermal growth factor receptor 3 (ERBB3) plasmid. The effects of overexpression of MALAT1, knockdown of ERBB3 and/or osimertinib on the proliferation of HCC827 cells were evaluated by 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2H tetrazolium bromide (MTT) assay. Cell apoptosis induced by MALAT1 overexpression, knockdown of ERBB3 and/or osimertinib treatment were analyzed by flow cytometry analysis. The expressions of EGFR and ERBB3 signal pathway related proteins in HCC827 cells treated with overexpression of MALAT1, knockdown of ERBB3 and/or osimertinib treatment were detected by western blot. Results: The MTT assay showed that sensitivity to osimertinib of HCC827/MALAT1 cells were significantly repressed. The 50% inhibitive concentration (IC₅₀) of osimertinib >4 000 nmol/L in HCC827/MALAT1 cells. However, knockdown of ERBB3 facilitated the anti-proliferation effect of osimertinib, and the IC₅₀ of osimertinib in shRNA-ERBB3 cells was (17.27±3.21) nmol/L. The results of flow cytometry analysis showed that the apoptotic rate of HCC827/MALAT1 cells induced by 10 nmol/L osimertinib was (8.38±0.92)%, significantly lower than (27.17±5.83)% of knockdown of ERBB3 (P<0.01). Western blotting showed that the expression of p-ERBB3, p-AKT and p-extracellular regulated protein kinases (ERK) in HCC827/MALAT1 cells was markedly up-regulated, while the expression of p-epithelial growth factor receptor (EGFR) was inhibited. The expressions of p-ERBB3, p-AKT and p-ERK were marginally affected by osimertinb. However, osimertinib downregulated the expressions of p-EGFR, p-ERBB3, p-AKT and p-ERK in ERBB3 deleted cells. Conclusions MALAT1 confers resistance to osimertinb in HCC827 cells by activating of the ERBB3/PI3K/AKT and ERBB3/MAPK/ERK signaling pathways.

Objective: To investigate the relationship of procollagen-lysine 2-oxoglutarate 5-dioxygenase 2 (PLOD2) expression and the clinical characteristics of osteosarcoma, and explore the potential mechanism of tumour metastasis promoted by PLOD2. Methods: The expression of PLOD2 in osteosarcoma tissues and paired adjacent tissues were detected by immunohistochemistry and qRT-PCR. Correlation of PLOD2 expression in osteosarcoma with the clinical pathologic features was analyzed by Chi square test and Kaplan-Meier analysis.Fibrillar collagen formation and collagen deposition in the tumor tissues were detected by picrosirius red staining. We transfected U-2OS cells with LV-vector, LV-over/PLOD2, sh-NC and sh-PLOD2. The expression of PLOD2 was detected by qRT-PCR. The impact of POLD2 on U-2OS cell invasion was determined by wound-healing assay and Transwell migration assay. The expressions of PLOD2/FAK/JAK2-STAT3 signal pathway related proteins were detected by western blotting. Results: The high expression level of PLOD2 in osteosarcoma tissues was 72.5%, significantly higher than 0% in paired adjacent noncancerous tissues (P<0.01), the expression of PLOD2 was positively correlated with lymph node metastasis, pulmonary metastasis and poor outcome (P<0.01). The same results were also observed in qRT-PCR assay. The median survival time of patients with high expression of PLOD2 protein was 13 months, significantly shorter than 32 months of patients with low expression of PLOD2 (P<0.05). The result of picrosirius red staining showed that the percentage of collagen fiber deposition in the osteosarcoma tissue with high level of PLOD2 was (74.43+ 9.63)%, significantly higher than (9.67±1.28)% in tissue with low expression of PLOD2 (P<0.001). The result of wound-healing and Transwell migration assay showed that over-expression of PLOD2 markedly promoted the invasion, however, knockdown of PLOD2 suppressed the invasion of U-2OS cells (both P<0.01). The result of western blotting showed that over-expression of PLOD2 significantly increased the expression levels of p-FAK, p-JAK2, p-STAT3, but knockdown PLOD2 decreased the levels of p-FAK, p-JAK2, p-STAT3 in U-2OS cells. Conclusions Up-regulation of PLOD2 in osteosarcoma is correlated with lymphatic and distant metastasis. PLOD2 promotes invasion and metastasis of osteosarcoma might through FAK/JAK2-STAT3 signal pathway.

Objective To test the effect of metastasis associated in lung adenocarcinoma transcript 1 ( MALAT1) and／or osimertinib on the proliferation and apoptosis of HCC827 cells, and explore the potential mechanism of MALAT1 induced resistance to osimertinib. Methods We transfected HCC827 cells with LV?vector or LV?over／MALAT1. Stable transfected cells ( HCC827／Vector, HCC827／MALAT1) were selected by adding puromycin. HCC827／MALAT1 cells were further transfected with the shRNA?negative control ( NC) or shRNA?human epidermal growth factor receptor 3 ( ERBB3 ) plasmid. The effects of overexpression of MALAT1, knockdown of ERBB3 and／or osimertinib on the proliferation of HCC827 cells were evaluated by 3?(4,5?dimethyl?2?thiazolyl)?2,5?diphenyl?2H tetrazolium bromide ( MTT) assay. Cell apoptosis induced by MALAT1 overexpression, knockdown of ERBB3 and／or osimertinib treatment were analyzed by flow cytometry analysis. The expressions of EGFR and ERBB3 signal pathway related proteins in HCC827 cells treated with overexpression of MALAT1, knockdown of ERBB3 and／or osimertinib treatment were detected by western blot. Results The MTT assay showed that sensitivity to osimertinib of HCC827／MALAT1 cells were significantly repressed. The 50% inhibitive concentration (IC50 ) of osimertinib ＞4 000 nmol／L in HCC827／MALAT1 cells.However, knockdown of ERBB3 facilitated the anti?proliferation effect of osimertinib, and the IC50 of osimertinib in shRNA?ERBB3 cells was (17.27±3.21) nmol／L. The results of flow cytometry analysis showed that the apoptotic rate of HCC827／MALAT1 cells induced by 10 nmol／L osimertinib was (8.38±0.92)%, significantly lower than ( 27.17± 5.83)% of knockdown of ERBB3 ( P＜0.01). Western blotting showed that the expression of p?ERBB3, p?AKT and p?extracellular regulated protein kinases ( ERK) in HCC827／MALAT1 cells was markedly up?regulated, while the expression of p?epithelial growth factor receptor (EGFR) was inhibited. The expressions of p?ERBB3, p?AKT and p?ERK were marginally affected by osimertinb. However, osimertinib downregulated the expressions of p?EGFR, p?ERBB3, p?AKT and p?ERK in ERBB3 deleted cells. Conclusions MALAT1 confers resistance to osimertinb in HCC827 cells by activating of the ERBB3／PI3K／AKT and ERBB3／MAPK／ERK signaling pathways.

Objective To investigate the relationship of procollagen?lysine 2?oxoglutarate 5?dioxygenase 2 ( PLOD2 ) expression and the clinical characteristics of osteosarcoma, and explore the potential mechanism of tumour metastasis promoted by PLOD2. Methods The expression of PLOD2 in osteosarcoma tissues and paired adjacent tissues were detected by immunohistochemistry and qRT?PCR. Correlation of PLOD2 expression in osteosarcoma with the clinical pathologic features was analyzed by Chi square test and Kaplan?Meier analysis.Fibrillar collagen formation and collagen deposition in the tumor tissues were detected by picrosirius red staining. We transfected U?2OS cells with LV?vector, LV?over／PLOD2, sh?NC and sh?PLOD2. The expression of PLOD2 was detected by qRT?PCR. The impact of POLD2 on U?2OS cell invasion was determined by wound?healing assay and Transwell migration assay. The expressions of PLOD2／FAK／JAK2?STAT3 signal pathway related proteins were detected by western blotting. Results The high expression level of PLOD2 in osteosarcoma tissues was 72.5%, significantly higher than 0% in paired adjacent noncancerous tissues ( P＜0.01), the expression of PLOD2 was positively correlated with lymph node metastasis, pulmonary metastasis and poor outcome (P＜0.01). The same results were also observed in qRT?PCR assay. The median survival time of patients with high expression of PLOD2 protein was 13 months, significantly shorter than 32 months of patients with low expression of PLOD2 ( P＜0.05). The result of picrosirius red staining showed that the percentage of collagen fiber deposition in the osteosarcoma tissue with high level of PLOD2 was (74.43+9.63)%, significantly higher than ( 9.67± 1.28)% in tissue with low expression of PLOD2 (P＜0.001).The result of wound?healing and Transwell migration assay showed that over?expression of PLOD2 markedly promoted the invasion, however, knockdown of PLOD2 suppressed the invasion of U?2OS cells ( both P＜0.01).The result of western blotting showed that over?expression of PLOD2 significantly increased the expression levels of p?FAK, p?JAK2, p?STAT3, but knockdown PLOD2 decreased the levels of p?FAK, p?JAK2, p?STAT3 in U?2OS cells. Conclusions Up?regulation of PLOD2 in osteosarcoma is correlated with lymphatic and distant metastasis. PLOD2 promotes invasion and metastasis of osteosarcoma might through FAK／JAK2?STAT3 signal pathway.

Objective To test the effect of metastasis associated in lung adenocarcinoma transcript 1 ( MALAT1) and／or osimertinib on the proliferation and apoptosis of HCC827 cells, and explore the potential mechanism of MALAT1 induced resistance to osimertinib. Methods We transfected HCC827 cells with LV?vector or LV?over／MALAT1. Stable transfected cells ( HCC827／Vector, HCC827／MALAT1) were selected by adding puromycin. HCC827／MALAT1 cells were further transfected with the shRNA?negative control ( NC) or shRNA?human epidermal growth factor receptor 3 ( ERBB3 ) plasmid. The effects of overexpression of MALAT1, knockdown of ERBB3 and／or osimertinib on the proliferation of HCC827 cells were evaluated by 3?(4,5?dimethyl?2?thiazolyl)?2,5?diphenyl?2H tetrazolium bromide ( MTT) assay. Cell apoptosis induced by MALAT1 overexpression, knockdown of ERBB3 and／or osimertinib treatment were analyzed by flow cytometry analysis. The expressions of EGFR and ERBB3 signal pathway related proteins in HCC827 cells treated with overexpression of MALAT1, knockdown of ERBB3 and／or osimertinib treatment were detected by western blot. Results The MTT assay showed that sensitivity to osimertinib of HCC827／MALAT1 cells were significantly repressed. The 50% inhibitive concentration (IC50 ) of osimertinib ＞4 000 nmol／L in HCC827／MALAT1 cells.However, knockdown of ERBB3 facilitated the anti?proliferation effect of osimertinib, and the IC50 of osimertinib in shRNA?ERBB3 cells was (17.27±3.21) nmol／L. The results of flow cytometry analysis showed that the apoptotic rate of HCC827／MALAT1 cells induced by 10 nmol／L osimertinib was (8.38±0.92)%, significantly lower than ( 27.17± 5.83)% of knockdown of ERBB3 ( P＜0.01). Western blotting showed that the expression of p?ERBB3, p?AKT and p?extracellular regulated protein kinases ( ERK) in HCC827／MALAT1 cells was markedly up?regulated, while the expression of p?epithelial growth factor receptor (EGFR) was inhibited. The expressions of p?ERBB3, p?AKT and p?ERK were marginally affected by osimertinb. However, osimertinib downregulated the expressions of p?EGFR, p?ERBB3, p?AKT and p?ERK in ERBB3 deleted cells. Conclusions MALAT1 confers resistance to osimertinb in HCC827 cells by activating of the ERBB3／PI3K／AKT and ERBB3／MAPK／ERK signaling pathways.

Objective To investigate the relationship of procollagen?lysine 2?oxoglutarate 5?dioxygenase 2 ( PLOD2 ) expression and the clinical characteristics of osteosarcoma, and explore the potential mechanism of tumour metastasis promoted by PLOD2. Methods The expression of PLOD2 in osteosarcoma tissues and paired adjacent tissues were detected by immunohistochemistry and qRT?PCR. Correlation of PLOD2 expression in osteosarcoma with the clinical pathologic features was analyzed by Chi square test and Kaplan?Meier analysis.Fibrillar collagen formation and collagen deposition in the tumor tissues were detected by picrosirius red staining. We transfected U?2OS cells with LV?vector, LV?over／PLOD2, sh?NC and sh?PLOD2. The expression of PLOD2 was detected by qRT?PCR. The impact of POLD2 on U?2OS cell invasion was determined by wound?healing assay and Transwell migration assay. The expressions of PLOD2／FAK／JAK2?STAT3 signal pathway related proteins were detected by western blotting. Results The high expression level of PLOD2 in osteosarcoma tissues was 72.5%, significantly higher than 0% in paired adjacent noncancerous tissues ( P＜0.01), the expression of PLOD2 was positively correlated with lymph node metastasis, pulmonary metastasis and poor outcome (P＜0.01). The same results were also observed in qRT?PCR assay. The median survival time of patients with high expression of PLOD2 protein was 13 months, significantly shorter than 32 months of patients with low expression of PLOD2 ( P＜0.05). The result of picrosirius red staining showed that the percentage of collagen fiber deposition in the osteosarcoma tissue with high level of PLOD2 was (74.43+9.63)%, significantly higher than ( 9.67± 1.28)% in tissue with low expression of PLOD2 (P＜0.001).The result of wound?healing and Transwell migration assay showed that over?expression of PLOD2 markedly promoted the invasion, however, knockdown of PLOD2 suppressed the invasion of U?2OS cells ( both P＜0.01).The result of western blotting showed that over?expression of PLOD2 significantly increased the expression levels of p?FAK, p?JAK2, p?STAT3, but knockdown PLOD2 decreased the levels of p?FAK, p?JAK2, p?STAT3 in U?2OS cells. Conclusions Up?regulation of PLOD2 in osteosarcoma is correlated with lymphatic and distant metastasis. PLOD2 promotes invasion and metastasis of osteosarcoma might through FAK／JAK2?STAT3 signal pathway.

Objective: To study the experience of famous old Traditional Chinese Medicine doctors of Jiangsu, Zhejiangand Shanghai in applying medicinal paste according to the deconstruction analysis of the medical cases formed by miningthe linked data of original cases on the basis of TCM features. Methods: To apply a deconstruction analysis to thesedoctors. medical cases with the quantifiable trend data on the data processing platform of Medcase. Results: This studycollected 472 medical visits, 250 pathogenesis elements, 400 therapies and 529 kinds of herbs. Conclusion: The medicinal pastes are applied by these famous doctors to treat commonly seen consumptive diseases, namely to intervenesub-health status. The chief core indications are lassitude, poor sleep, lumbar soreness and poor appetite; the relatedtongue and pulse conditions are thin or white tongue coating, red or pale tongue proper, and a thready or wiry pulse; thechief pathogenesis elements are deficiency of the liver and kidney, and deficiency of the spleen and kidney; thefrequently used herbs are Radix Astragali seu Hedysari, Rhizoma Atractylodis Macrocephalae, Poria, Radix Angelicae Sinensis, Radix Paeoniae, Radix Glycyrrhizae, Radix Rehmanniae Preparata, etc. The prescriptions for medicinal pastesare made by modifying several basic formulas including Sijunzi Decoction, Siwu Decoction, Bazhen Decoction, DangguiBuxue Decoction, Liuwei Dihuang Pills, and Shengmai Decoction. The common excipients for pastes are Ejiao, rockcandy, Guibanjiao, Lujiaojio, etc. The guide for making the pastes are combined therapies, such as regulating both qi andblood, tonifying both yin and yang, and treating both deficiency and excess.