In 2020,the Pharmaceutical Service Committee of the Chinese Pharmaceutical Association,the Respiratory Disease Branch of the Chinese Medical Association,and the Respiratory Physician Branch of the Chinese Medical Doctor Associ-ation jointly established the"Cough and Wheezing Pharmaceutical Care Clinic(CWPC)program".The specification on CWPC construction provides guidance and reference for improving the quality of cough and wheezing pharmaceutical care services.The Chinese Pharmaceutical Association Specifications on Cough and Wheeze Pharmaceutical Care Clinic construction covered two as-pects,the construction and service of CWPC,respectively.With the analysis of relevant standards,policies and regulations,tech-nical specifications,literature,and other materials,the specification standardized 19 key elements in the organization's construc-tion,service process,quality management,and evaluation improvement.This paper mainly analyzed the construction process and content of the Chinese Pharmaceutical Association Specifications on Cough and Wheeze Pharmaceutical Care Clinic construction,to strengthen the publicity and implementation of the specification,deepen the understanding of the specification content,and pro-mote the implementation of the specification.

Objective:To establish a complete system for the isolation, purification, identification, and culture of Kaposiform hemangioendothelioma-derived endothelial cells (KHE-ECs), to analyze the phenotype of KHE-ECs, and to explore the possibility of establishing a KHE-EC bank.Methods:A novel digestion solution for KHE tumors (patent number: CN202410500224.2) was formulated using collection fluid, Liberase TM and dispase stock solutions, and was used to process tumor tissues to obtain cells. High-purity KHE-ECs were purified using CD31 + immunomagnetic beads. The EGM-2 complete medium containing 10% fetal bovine serum and 2% penicillin-streptomycin solution was employed for cell culture. To verify the characteristics of KHE-ECs, immunofluorescence assay was conducted to determine the expression of endothelial cell-specific markers CD31 and CD34, KHE disease markers podoplanin (D2-40), prospero-related homeobox 1 (Prox-1), and lymphatic vessel endothelial hyaluronan receptor 1 (LYVE1), as well as an infantile hemangioma-specific diagnostic marker glucose transporter 1 (GLUT-1). Human umbilical vein endothelial cells (HUVECs) served as controls for the phenotype analysis of KHE-ECs, including cell viability, cytoskeleton, proliferation, migration, invasion, tube formation, and sprouting ability. Results:Primary cells were successfully isolated from KHE tumor tissues, and high-purity KHE-ECs were obtained by using CD31 + immunomagnetic beads. The cells exhibited typical spindle-shaped morphology and an adherent growth pattern. Immunofluorescence assay showed that KHE-ECs expressed CD31, CD34, D2-40, Prox-1, and LYVE1, but did not express GLUT-1. There were significant differences in cell morphology, cell viability, and cytoskeletal structures between KHE-ECs and HUVECs. Additionally, the KHE-EC group showed significantly increased percentages of proliferative cells (29.1% ± 2.5%), numbers of migratory cells (114.3 ± 9.4) and invasive cells (110.0 ± 6.1), tube length (32 121.0 ± 892.0 μm), and number of sprouting cells (25.0 ± 3.6) compared with the HUVEC group (13.0% ± 2.2%, 38.0 ± 3.6, 35.3 ± 2.3, 25 345.0 ± 448.1 μm, 5.0 ± 1.0, respectively, all P ≤ 0.001) . Conclusion:An innovative digestion solution specifically for KHE tumors was formulated for the first time, and high-purity and well-growing KHE-EC strains were successfully isolated and purified by using the novel digestion solution in combination with CD31 + immunomagnetic beads, providing a stable and reliable cell source for subsequent experimental studies on KHE and laying the foundation for establishing a KHE-EC bank.

Objective:To establish a complete system for the isolation, purification, identification, and culture of Kaposiform hemangioendothelioma-derived endothelial cells (KHE-ECs), to analyze the phenotype of KHE-ECs, and to explore the possibility of establishing a KHE-EC bank.Methods:A novel digestion solution for KHE tumors (patent number: CN202410500224.2) was formulated using collection fluid, Liberase TM and dispase stock solutions, and was used to process tumor tissues to obtain cells. High-purity KHE-ECs were purified using CD31 + immunomagnetic beads. The EGM-2 complete medium containing 10% fetal bovine serum and 2% penicillin-streptomycin solution was employed for cell culture. To verify the characteristics of KHE-ECs, immunofluorescence assay was conducted to determine the expression of endothelial cell-specific markers CD31 and CD34, KHE disease markers podoplanin (D2-40), prospero-related homeobox 1 (Prox-1), and lymphatic vessel endothelial hyaluronan receptor 1 (LYVE1), as well as an infantile hemangioma-specific diagnostic marker glucose transporter 1 (GLUT-1). Human umbilical vein endothelial cells (HUVECs) served as controls for the phenotype analysis of KHE-ECs, including cell viability, cytoskeleton, proliferation, migration, invasion, tube formation, and sprouting ability. Results:Primary cells were successfully isolated from KHE tumor tissues, and high-purity KHE-ECs were obtained by using CD31 + immunomagnetic beads. The cells exhibited typical spindle-shaped morphology and an adherent growth pattern. Immunofluorescence assay showed that KHE-ECs expressed CD31, CD34, D2-40, Prox-1, and LYVE1, but did not express GLUT-1. There were significant differences in cell morphology, cell viability, and cytoskeletal structures between KHE-ECs and HUVECs. Additionally, the KHE-EC group showed significantly increased percentages of proliferative cells (29.1% ± 2.5%), numbers of migratory cells (114.3 ± 9.4) and invasive cells (110.0 ± 6.1), tube length (32 121.0 ± 892.0 μm), and number of sprouting cells (25.0 ± 3.6) compared with the HUVEC group (13.0% ± 2.2%, 38.0 ± 3.6, 35.3 ± 2.3, 25 345.0 ± 448.1 μm, 5.0 ± 1.0, respectively, all P ≤ 0.001) . Conclusion:An innovative digestion solution specifically for KHE tumors was formulated for the first time, and high-purity and well-growing KHE-EC strains were successfully isolated and purified by using the novel digestion solution in combination with CD31 + immunomagnetic beads, providing a stable and reliable cell source for subsequent experimental studies on KHE and laying the foundation for establishing a KHE-EC bank.

In 2020,the Pharmaceutical Service Committee of the Chinese Pharmaceutical Association,the Respiratory Disease Branch of the Chinese Medical Association,and the Respiratory Physician Branch of the Chinese Medical Doctor Associ-ation jointly established the"Cough and Wheezing Pharmaceutical Care Clinic(CWPC)program".The specification on CWPC construction provides guidance and reference for improving the quality of cough and wheezing pharmaceutical care services.The Chinese Pharmaceutical Association Specifications on Cough and Wheeze Pharmaceutical Care Clinic construction covered two as-pects,the construction and service of CWPC,respectively.With the analysis of relevant standards,policies and regulations,tech-nical specifications,literature,and other materials,the specification standardized 19 key elements in the organization's construc-tion,service process,quality management,and evaluation improvement.This paper mainly analyzed the construction process and content of the Chinese Pharmaceutical Association Specifications on Cough and Wheeze Pharmaceutical Care Clinic construction,to strengthen the publicity and implementation of the specification,deepen the understanding of the specification content,and pro-mote the implementation of the specification.

Objective:To analyze the impact of cecal ligation and puncture (CLP)-induced sepsis on the proliferation and differentiation of intestinal epithelial cells.Methods:① Animal experiment: sixteen male C57BL/6 mice were divided into sham operation group (Sham group) and CLP-induced sepsis model group (CLP group) by random number table method, with 8 mice in each group. After 5 days of operation, the jejunal tissues were taken for determination of leucine-rich-repeat-containing G-protein-coupled receptor 5 (LGR5) and intestinal alkaline phosphatase (IAP) by polymerase chain reaction (PCR). The translation of LGR5 was detected by Western blotting. The expression of proliferating cell nuclear antigen (Ki67) was analyzed by immunohistochemistry. IAP level was detected by modified calcium cobalt staining and colorimetry. Immunofluorescence staining was used to detect the expression of Paneth cell marker molecule lysozyme 1 (LYZ1) and goblet cell marker molecule mucin 2 (MUC2). ② Cell experiment: IEC6 cells in logarithmic growth stage were divided into blank control group and lipopolysaccharide (LPS) group (LPS 5 μg/mL). Twenty-four hours after treatment, PCR and Western blotting were used to analyze the transcription and translation of LGR5. The proliferation of IEC6 cells were detected by 5-ethynyl-2'-deoxyuridine (EdU) staining. The transcription and translation of IAP were detected by PCR and colorimetric method respectively.Results:① Animal experiment: the immunohistochemical results showed that the positive rate of Ki67 staining in the jejunal tissue of CLP group was lower than that of Sham group [(41.7±2.5)% vs. (48.7±1.4)%, P = 0.01]. PCR and Western blotting results showed that there were no statistical differences in the mRNA and protein expressions of LGR5 in the jejunal tissue between the CLP group and Sham group (Lgr5 mRNA: 0.7±0.1 vs. 1.0±0.2, P = 0.11; LGR5/β-actin: 0.83±0.17 vs. 0.68±0.19, P = 0.24). The mRNA (0.4±0.1 vs. 1.0±0.1, P < 0.01) and protein (U/g: 47.3±6.0 vs. 73.1±15.3, P < 0.01) levels of IAP in the jejunal tissue were lower in CLP group. Immunofluorescence saining analysis showed that the expressions of LYZ1 and MUC2 in the CLP group were lower than those in the Sham group. ②Cell experiment: PCR and Western blotting results showed that there was no significant difference in the expression of LGR5 between the LPS group and the blank control group (Lgr5 mRNA: 0.9±0.1 vs. 1.0±0.2, P = 0.33; LGR5/β-actin: 0.71±0.18 vs. 0.69±0.04, P = 0.81). The proliferation rate of IEC6 cells in the LPS group was lower than that in the blank control group, but there was no significant difference [positivity rate of EdU: (40.5±3.8)% vs. (46.5±3.6)%, P = 0.11]. The mRNA (0.5±0.1 vs. 1.0±0.2, P < 0.01) and protein (U/g: 15.0±4.0 vs. 41.2±10.4, P < 0.01) of IAP in the LPS group were lower than those in the blank control group. Conclusion:CLP-induced sepsis inhibits the proliferation and differentiation of intestinal epithelial cells, impairing the self-renewal ability of intestinal epithelium.

Objective:To analyze the clinical features, differential diagnosis, treatment and prognosis of complex lymphatic malformations.Methods:The clinical data of patients with complex lymphatic malformation were retrospectively analyzed from April 2010 to April 2022 in the Multidisciplinary Outpatient Department of the Vascular Disease Team of West China Hospital, Sichuan University. All patients were diagnosed with complex lymphatic malformation after consultation with multidisciplinary experts in pediatric surgery, radiology, plastic surgery, pathology, rehabilitation and other departments. The clinical manifestations, blood routine, coagulation function, magnetic resonance imaging and treatment methods of the patients were analyzed. According to the follow-up and disease results, the patients were divided into improvement, stability, progress and death.Results:A total of 18 patients with complex lymphatic malformations were included in the study, including 6 males and 12 females. The age of first diagnosis ranged from 1 month to 29 years old, and the median age was 2.5 years old. Patients were followed up and treated for 0.4 to 12.0 years, with an average follow-up of 3.5 years. Ten patients had pleural and pericardial effusion; 15 patients had visceral involvement which showed multifocal changes in imaging examinations; 9 cases were accompanied by bone destruction, which in Gorham-Stout disease patients broke through the cortex while in generalized lymphatic anomalies it did not; 14 patients had various degrees of coagulation abnormalities, of which 8 patients with severe coagulation dysfunction were all diagnosed as kaposiform lymphangiomatosis. Of the 18 patients, one kaposiform lymphangiomatosis patient died; six patients progressed; eight patients were stable; and three patients improved.Conclusion:The clinical characteristics of patients with complex lymphatic malformations are systemic, diverse and complex. The clinical symptoms of patients with diffuse lymphatic malformation accompanied by involvement of bone and multiple internal organs, chest and abdominal effusion, and coagulation dysfunction should be considered as complex lymphatic malformation. However, due to overlapping clinical characteristics of each subtypes, it is difficult to distinguish patients with complex lymphatic malformation, and the curative effect and prognosis are poor. Precision targeted drugs are the future research direction for the treatment of such diseases.

Objective:To analyze the clinical features, differential diagnosis, treatment and prognosis of complex lymphatic malformations.Methods:The clinical data of patients with complex lymphatic malformation were retrospectively analyzed from April 2010 to April 2022 in the Multidisciplinary Outpatient Department of the Vascular Disease Team of West China Hospital, Sichuan University. All patients were diagnosed with complex lymphatic malformation after consultation with multidisciplinary experts in pediatric surgery, radiology, plastic surgery, pathology, rehabilitation and other departments. The clinical manifestations, blood routine, coagulation function, magnetic resonance imaging and treatment methods of the patients were analyzed. According to the follow-up and disease results, the patients were divided into improvement, stability, progress and death.Results:A total of 18 patients with complex lymphatic malformations were included in the study, including 6 males and 12 females. The age of first diagnosis ranged from 1 month to 29 years old, and the median age was 2.5 years old. Patients were followed up and treated for 0.4 to 12.0 years, with an average follow-up of 3.5 years. Ten patients had pleural and pericardial effusion; 15 patients had visceral involvement which showed multifocal changes in imaging examinations; 9 cases were accompanied by bone destruction, which in Gorham-Stout disease patients broke through the cortex while in generalized lymphatic anomalies it did not; 14 patients had various degrees of coagulation abnormalities, of which 8 patients with severe coagulation dysfunction were all diagnosed as kaposiform lymphangiomatosis. Of the 18 patients, one kaposiform lymphangiomatosis patient died; six patients progressed; eight patients were stable; and three patients improved.Conclusion:The clinical characteristics of patients with complex lymphatic malformations are systemic, diverse and complex. The clinical symptoms of patients with diffuse lymphatic malformation accompanied by involvement of bone and multiple internal organs, chest and abdominal effusion, and coagulation dysfunction should be considered as complex lymphatic malformation. However, due to overlapping clinical characteristics of each subtypes, it is difficult to distinguish patients with complex lymphatic malformation, and the curative effect and prognosis are poor. Precision targeted drugs are the future research direction for the treatment of such diseases.

Objective:To analyze demographic and clinical characteristics of infantile hemangioma (IH) , and to explore related risk factors for IH.Methods:A multicenter case-control study was conducted. IH patients (case group) and healthy children (control group) were collected from West China Hospital of Sichuan University, West China Second University Hospital of Sichuan University and Yulin Community Central Hospital of Chengdu from October 2018 to December 2020. The data on patients′ demographic characteristics, and risk factors during their mothers′ pre-pregnancy, pregnancy and perinatal period were collected and retrospectively analyzed. Univariate and multivariate analyses were performed using binary logistic regression.Results:A total of 1 479 patients with IH and 1 086 healthy children were included in this study. There were 456 males and 1 023 females in the case group, with the age being 3.74 ± 2.82 months, and there were 359 males and 727 females in the control group, with the age being 3.95 ± 2.77 months. There was no significant difference in the gender ratio, age, ethnic composition, birth weight or birth height between the case group and control group (all P > 0.05) . IH lesions mostly affected the head and face (564 cases, 38.1%) , followed by the trunk (449 cases, 30.6%) and limbs (356 cases, 24.1%) . At the visit, 1 109 (75.0%) patients presented with proliferating IH, 1 059 (71.6%) with superficial IH, and 1 306 (88.3%) with focal IH. The IH lesion area ranged from 0.01 to 168.00 (6.24 ± 12.91) cm 2, and the segmental IH area ranged from 7.50 to 168.00 (32.17 ± 26.94) cm 2. Univariate logistic regression analysis showed some factors influencing the occurrence of IH (all P < 0.05) , including pre-pregnancy factors (delivery history and miscarriage history) , pregnancy factors (fetal distress, cord entanglement, history of threatened abortion, placenta previa, oligohydramnios, gestational hypothyroidism, gestational anemia, history of progesterone supplementation, history of thyroxine drug use, history of uterus myomas) , and perinatal factors (including fetal position, gestational weeks, premature rupture of membranes and preterm premature rupture of membranes) . Multivariate binary logistic regression adjusted analysis showed that fetal breech presentation, preterm birth, cord entanglement and history of thyroxine drug use during pregnancy did not influence the occurrence of IH (all P > 0.05) ; the delivery history was the strongest independent risk factor for IH (adjusted OR = 5.624, 95% CI: 4.275 to 7.398, P < 0.001) , and gestational hypothyroidism and history of uterus myomas were protective factors for IH. Conclusions:In this study, the average age of IH patients at visit was 4 months, skin lesions mostly occurred on the head and face, and most were superficial and focal in the proliferative stage. The occurrence and development of IH may be associated with placental diseases, hypoxia, maternal hormone levels during pregnancy, etc.

Objective:To investigate the similarities and differences in clinical manifestations, lesion features, treatment options and prognosis in patients with kaposiform hemangioendothelioma (KHE) and tufted angioma (TA).Methods:The clinical data of KHE patients and TA patients diagnosed and treated in West China Hospital of Sichuan University from August 2016 to June 2020 were retrospectively analyzed. The similarities and differences in gender, the age of onset, clinical features, complications, histopathology, imaging manifestations, treatment and prognosis of the two diseases were compared. The χ2 test or Fisher’s exact test were used to analyze the qualitative data. The Mann-Whitney U test was used to analyze the quantitative data of the two groups that did not conform to the normal distribution. P<0.05 was considered to be statistically significant. Results:A total of 217 patients were included, including 183 patients with KHE and 34 patients with TA. There was no significant difference between KHE and TA in male to female ratio ( χ2=0.44, P=0.510), the age of onset ( U=2 757.00, Z=-1.09, P=0.278) and the sites of lesion ( χ2=3.64, P=0.162). The mixed type of KHE was the most common, reaching 63.39% (116/183), while the superficial type of TA was the most common, reaching 88.24% (30/34). The lesion diameter of KHE patients was 6.30(4.40, 9.70) cm, which was larger than that of TA patients 2.95(2.05, 4.03) cm, and the difference was statistically significant ( U=967.50, Z=-6.38, P<0.001). KHE is more likely to involve skeletal muscle, and cause thrombocytopenia and severe fibrinogenopenia. KHE mainly involved the dermis, subcutaneous tissue and even deep muscles, and manifested as an infiltrative mass. Magnetic resonance imaging (MRI) showed mass diffuse with high signal on T 2 phase. TA was often a superficial lesion that only involved the subcutaneous fat layer and was lumpy. MRI showed that the high signal was confined to the subcutaneous fat layer. The total effective rate of KHE [KHE(85.79%) vs. TA(91.18%)] and the total effective rate of drug therapy [KHE(85.32%) vs. TA(95.65%)] were lower than those of TA. Conclusions:Compared with TA, KHE has a larger tumor diameter, higher invasiveness, higher risk of complications. In addition, treatment plan was more complicated and treatment response rate was lower in patients with KHE compared with those in patients with TA.

Objective:To investigate the similarities and differences in clinical manifestations, lesion features, treatment options and prognosis in patients with kaposiform hemangioendothelioma (KHE) and tufted angioma (TA).Methods:The clinical data of KHE patients and TA patients diagnosed and treated in West China Hospital of Sichuan University from August 2016 to June 2020 were retrospectively analyzed. The similarities and differences in gender, the age of onset, clinical features, complications, histopathology, imaging manifestations, treatment and prognosis of the two diseases were compared. The χ2 test or Fisher’s exact test were used to analyze the qualitative data. The Mann-Whitney U test was used to analyze the quantitative data of the two groups that did not conform to the normal distribution. P<0.05 was considered to be statistically significant. Results:A total of 217 patients were included, including 183 patients with KHE and 34 patients with TA. There was no significant difference between KHE and TA in male to female ratio ( χ2=0.44, P=0.510), the age of onset ( U=2 757.00, Z=-1.09, P=0.278) and the sites of lesion ( χ2=3.64, P=0.162). The mixed type of KHE was the most common, reaching 63.39% (116/183), while the superficial type of TA was the most common, reaching 88.24% (30/34). The lesion diameter of KHE patients was 6.30(4.40, 9.70) cm, which was larger than that of TA patients 2.95(2.05, 4.03) cm, and the difference was statistically significant ( U=967.50, Z=-6.38, P<0.001). KHE is more likely to involve skeletal muscle, and cause thrombocytopenia and severe fibrinogenopenia. KHE mainly involved the dermis, subcutaneous tissue and even deep muscles, and manifested as an infiltrative mass. Magnetic resonance imaging (MRI) showed mass diffuse with high signal on T 2 phase. TA was often a superficial lesion that only involved the subcutaneous fat layer and was lumpy. MRI showed that the high signal was confined to the subcutaneous fat layer. The total effective rate of KHE [KHE(85.79%) vs. TA(91.18%)] and the total effective rate of drug therapy [KHE(85.32%) vs. TA(95.65%)] were lower than those of TA. Conclusions:Compared with TA, KHE has a larger tumor diameter, higher invasiveness, higher risk of complications. In addition, treatment plan was more complicated and treatment response rate was lower in patients with KHE compared with those in patients with TA.