Objective To establish the UPLC fingerprint chromatogram combined with chemometric analysis for the quality evaluation of classical formula Linggui Zhugan Decoction.Methods SHIMADZU Shim-Pack GIST C18 column(100 mm×2.1 mm,2.0 μm)was used with acetonitrile-0.1%phosphoric acid aqueous solution as mobile phase,gradient elution;flow rate was 0.2 mL/min;the detection wavelength was 266 nm for the first 30 minutes and 235 nm for the last 36 minutes;the column temperature was 30℃.The UPLC fingerprint of Linggui Zhugan Decoction was established by Similarity Evaluation System for Chromatographic Fingerprint of TCM(2012.130723 version),and the common peak was determined and the similarity evaluation was carried out.Based on the peak area determination results of the common peak of the fingerprint,the quality of different batches of Linggui Zhugan Decoction was evaluated by chemometrics such as clustering analysis and principal component analysis.Results A total of 24 common peaks were confirmed and 14 components were identified by using reference substances.The similarity of 10 batches of Linggui Zhugan Decoction samples was greater than 0.950,which could be divided into two categories by chemometrics,and the principal component 1-4 were the main factors affecting its quality evaluation.OPLS-DA identified 6 differential markers.Conclusion The fingerprint research method established in the study is simple,reliable and reproducible.Through the method of fingerprint combined with chemometrics analysis,the differences between Linggui Zhugan Decoction from different origins of medicinal materials are identified,which provides a reference for the internal quality evaluation of Linggui Zhugan Decoction.

ObjectiveTo explore the correlation between dryness and energy metabolism of Atractylodis Rhizoma， and to analyze the difference of medicinal properties between Atractylodes lancea and A. chinensis. MethodA total of 110 healthy SD rats were randomly divided into 11 groups， including normal group， volatile oil of A. lancea 1-5 group （S1-S5 group， doses of 447， 473， 442， 489， 496 mg·kg^-1） and volatile oil of A. chinensis 1-5 group （N1-N5 group， doses of 197， 118， 281， 222， 185 mg·kg^-1）， the administration volume was 0.01 mL·g^-1 with intragastric administration for 21 days. Dryness effect of A. lancea and A. chinensis on rats was evaluated by comparing the body weight， drinking water volume， urine volume， whole blood viscosity and pathological sections of submandibular gland stained with hematoxylin-eosin （HE）. The expression of aquaporin 2 （AQP2） in rat kidney was measured by immunohistochemistry， the mRNA expressions of cytochrome C oxidase subunit 7A2 （COX7A2） and succinate dehydrogenase （SDH） complex subunit D （SDHD） in liver tissue were detected by real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）. The contents of SDH， lactate dehydrogenase （LDH） and sodium ion-potassium ion-adenosine triphosphatase （Na⁺-K⁺-ATPase） in rat plasma were determined by colorimetry. The quality of A. lancea and A. chinensis was evaluated by coefficient of variation method， and Pearson correlation coefficient was used to analyze the correlation between dryness and energy metabolism. ResultCompared with the normal group， the amounts of drinking water and urine in volatile oil of A. lancea group and volatile oil of A. chinensis group increased， and the submandibular gland acini atrophied， the whole blood viscosity of rats in the volatile oil of A. lancea group increased significantly （P<0.01）， the expression levels of COX7A2 and SDHD mRNA， the activities of SDH， LDH and Na⁺-K⁺-ATPase increased significantly （P<0.01）， and the expression of AQP2 in kidney decreased significantly （P<0.01）. Compared with the normal group， the expression level of COX7A2 mRNA， SDH activity and whole blood viscosity in the volatile oil of A. chinensis group increased （P<0.05， P<0.01）， the AQP2 and SDH mRNA expression levels， LDH and Na⁺-K⁺-ATPase activities had no significant difference. The comprehensive score analysis of each index showed that the effect of volatile oil of A. lancea on dryness and energy metabolism was stronger than that of volatile oil of A. chinensis， and there was a positive correlation between dryness index and energy metabolism index. ConclusionThe two indexes show that medicinal properties of A. lancea is stronger than that of A. chinensis， and energy metabolism is closely related to the dryness of Atractylodis Rhizoma. It is suggested that it is reasonable to evaluate the dryness effect of Atractylodis Rhizoma from the perspective of energy metabolism， which can further enrich the evaluation indexes of medicinal properties.

Objective:To explore the effect of family integrated care (FIC) combined with intelligent health education platform in health education for elderly patients with stroke.Methods:Eighty patients with stroke were included from the Department of Neurology of the Second Affiliated Hospital of Chongqing Medical University from June 2019 to May 2020. They were divided into the experimental group and the control group ( n=40, respectively) by the random number table method. Patients in the control group received routine FIC in the Department of Neurology. Additional intelligent health education platform was provided for the experimental group to carry out intensive education. The modified Barthel index scale, self-care compliance scale and hospital anxiety and depression scale (HADS) were used before and after the intervention to evaluate patient′s self-care ability, self-care compliance and the degree of anxiety and depression. Results:Finally, 39 patients in the experimental group (1 case lost to followed-up) and 37 patients in the control group (3 cases lost to followed-up) completed the study. After the intervention, the scores of the modified Barthel index and self-care compliance scale of the experimental group were significantly higher than those of the control group [(65.9±12.6) vs (60.1±11.9) points, (78.2±13.6) vs (71.4±14.6) points], the HADS score was significantly lower than the control group [(15.18±2.46) vs (19.46±2.40) points] (all P<0.05). The scores of the modified Barthel index scale and self-care compliance scale of the experimental group after the intervention were significantly higher than those before the intervention [(65.9±12.6) vs (40.8±12.5) points, (78.2±13.6) vs (54.6±15.9) points] (all P<0.05), the HADS score was significantly lower than before the intervention [(15.18±2.46) vs (21.74±3.52) points] (all P<0.05). Similarly,the scores of the modified Barthel index and self-care compliance scale of the control group after the intervention were significantly higher than before the intervention [(60.1±11.9) vs (41.6±9.72) points, (71.4±14.6) vs (54.3±14.8) points], the HADS score was significantly lower than before the intervention [(19.46±2.40) vs (21.38±3.09) points] (all P<0.05). Conclusion:FIC combined with intelligent health education platform can significantly improve the self-care ability and self-nursing compliance of elderly stroke patients, and markedly improve the degree of depression and anxiety.

Objective:To explore the role of parental origin verification in chromosomal microarray analysis (CMA) on the determination of the clinical significance of copy number variations (CNVs).Methods:This retrospective study collected clinical information from 73 core families who underwent prenatal diagnosis at Peking University First Hospital from November 2017 to December 2019. Indications for prenatal diagnosis included ultrasound abnormality in 54 cases (including 12 with thickened nuchal translucency (≥2.5 mm), four with fetal growth restriction, seven with abnormal pregnancy history, and 31 with isolated ultrasound abnormality), NIPT indicated high-risk in four cases, advanced age in nine cases, abnormal pregnancy history alone in three cases, intrauterine death in two cases and one with maternal mental retardation. Genomic DNA of amniotic fluid sample, chorionic villi, cord blood, fetal tissues, and fetal heart blood were extracted using genomic DNA extraction kit. The CNVs of prenatal samples in 73 subjects were analyzed using array-based comparative genomic hybridization (array-CGH) analysis and single nucleotide polymorphism array (SNP-array). Peripheral blood DNA of the couples, and relevant families if necessary, were collected and analyzed in the same way. The results of parental origin detection in CMA were summarized.Results:A total of 76 CNVs were detected in these 73 samples, out of which nine were pathogenic and parental origin detection revealed that six were de novo, two were maternally, and one was paternally inherited; six CNVs were likely pathogenic, including three de novo, two maternally inherited and one paternally inherited; 20 CNVs were variants of uncertain significance, including five paternally inherited, three maternally inherited and 12 de novo; 41 CNVs were likely benign, among which 38 were inherited from parents with normal phenotype. Conclusions:Parental origin verification plays an important role in explaining the clinical significance of detected fetal CNVs and thereby can help to analyze its clinical effect and reproductive risk.

OBJECTIVE: To explore the genetic basis for a pedigree affected with Charcot-Marie-Tooth (CMT) disease through high-throughput sequencing. METHODS: Potential variants of the genes associated with CMT were screened by next-generation sequencing (NGS) of the members of the pedigree. RESULTS: NGS has revealed that the two affected sisters both harbored homozygous c.1A>G variant of the GDAP1 gene, which caused replacement of the first amino acid Methionine by Valine (p.Met1Val). Their parents were both carriers of the heterozygous c.1A>G variant. The variant was unreported previously and has an extremely low frequency in the population. Meanwhile, one of the sisters and the mother also carried heterozygous c.710A>T variant of the BAG3 gene. CONCLUSION The homozygous c.1A>G variant of the GDAP1 gene probably underlay the CMT in both children. Above result has enabled clinical diagnosis and genetic counseling for this pedigree.
Adaptor Proteins, Signal Transducing/genetics* ; Apoptosis Regulatory Proteins/genetics* ; Charcot-Marie-Tooth Disease/genetics* ; Child ; Female ; Fibula/abnormalities* ; Homozygote ; Humans ; Mutation ; Nerve Tissue Proteins/genetics* ; Pedigree

Objective:To compare the efficacy of the combination of abidol, lopinavir/ritonavir plus recombinant interferon α-2b (rIFNα-2b) and the combination of lopinavir/ritonavir plus rIFNα-2b for patients with COVID-19 in Zhejiang province.Methods:A multicenter prospective study was carried out to compare the efficacy of triple combination antiviral therapy and dual combination antiviral therapy in 15 medical institutions of Zhejiang province during January 22 to February 16, 2020. All patients were treated with rIFNα-2b (5 million U, 2 times/d) aerosol inhalation, in addition 196 patients were treated with abidol (200 mg, 3 times/d) + lopinavir/ritonavir (2 tablets, 1 time/12 h) (triple combination group) and 41 patients were treated with lopinavir/ritonavir (2 tablets, 1 time/12 h) (dual combination group). The patients who received triple combination antiviral therapy were further divided into three subgroups: <48 h, 3-5 d and >5 d according the time from the symptom onset to medication starting. The therapeutic efficacy was compared between triple combination group and dual combination group, and compared among 3 subgroups of patients receiving triple combination antiviral therapy. SPSS 17.0 software was used to analyze the data.Results:The virus nucleic acid-negative conversion time in respiratory tract specimens was (12.2±4.7) d in the triple combination group, which was shorter than that in the dual combination group [(15.0±5.0) d] ( t=6.159, P<0.01). The length of hospital stay in the triple combination group [12.0 (9.0, 17.0) d] was also shorter than that in the dual combination group [15.0 (10.0, 18.0) d] ( H=2.073, P<0.05). Compared with the antiviral treatment which was started within after the symptom onset of in the triple combination group, the time from the symptom onset to the viral negative conversion was 13.0 (10.0, 17.0), 17.0 (13.0, 22.0) and 21.0 (18.0, 24.0) d in subgroups of 48 h, 3-5 d and >5 d, respectively ( Z=32.983, P<0.01), while the time from antiviral therapy to viral negative conversion was (11.8±3.9), (13.5±5.1) and (11.2±4.3) d, respectively( Z=6.722, P<0.05). Conclusions:The triple combination antiviral therapy of abidol, lopinavir/litonavir and rIFNα-2b shows shorter viral shedding time and shorter hospitalization time, compared with the dual combination antiviral therapy; and the earlier starting triple combination antiviral therapy will result in better antiviral efficacy.

Objective: Comparing the benefit of Abidor, lopinavir/ritonavir and recombinant interferon α-2b triple combination antiviral therapy and lopinavir/ritonavir and interferon dual combination antiviral therapy to hospitalized novel coronavirus pneumonia 2019 in Zhejiang province. Methods: A multi-center prospective study was carried out to compare the effect of triple combination antiviral therapy with dual combination antiviral therapy in 15 medical institutions of Zhejiang Province. All patients were treated with recombinant interferon α-2b (5 million U, 2 times/d) aerosol inhalation. 196 patients were treated with abidol (200 mg, 3 times/d) + lopinavir / ritonavir (2 tablets, 1 time/12 h) as the triple combination antiviral treatment group. 41 patients were treated with lopinavir / ritonavir (2 tablets, 1 time/12 h) as the dual combination antiviral treatment group. The patients who received triple combination antiviral therapy were divided into three groups: within 48 hours, 3-5 days and > 5 days after the symptom onset. To explore the therapeutic effects of triple combination antiviral drugs and dual combination antiviral drugs, as well as triple combination antiviral drugs with different antiviral initiate time. SPSS17.0 software was used to analyze the data. Results: The time of virus nucleic acid turning negative was (12.2 ± 4.7) days in the triple combination antiviral drug group, which was shorter than that in the dual combination antiviral drug group [(15.0 ± 5.0) days] (t = 6.159, P < 0.01 ). The length of hospital stay [12 (9, 17) d] in the triple combination antiviral drug group was also shorter than that in the dual combination antiviral drug group [15 (10, 18) d] (H = 2.073, P < 0.05). Comparing the antiviral treatment which was started within 48 hours, 3-5 days and > 5 days after the symptom onset of triple combination antiviral drug group, the time from the symptom onset to the negative of viral shedding was 13 (10,16.8), 17 (13,22) and 21 (18-24) days respectively (Z = 32.983, P < 0.01), and the time from antiviral therapy to the negative of viral shedding was (11.8±3.9) , (13.5±5.1) and (11.2±4.3) d. The differences among the three groups were statistically significant (Z=32.983 and 6.722, P<0.01 or<0.05). Conclusions The triple combination antiviral therapy of Abidor, Lopinavir/Litonavir and recombinant interferon α-2b showed shorter viral shedding time and hospitalization time compared with the dual combination antiviral therapy. The earlier the time to initiate triple antiviral treatment, the shorter the time of virus shedding.

OBJECTIVE： To provide reference for reducing the rate of outpatient/emergency intravenous infusion in paediatric and improving the safety of drug use in children. METHODS： The comprehensive intervention of outpatient/emergency intravenous infusion in paediatric by pharmacists of our hospital though multiple measures was introduced， such as education training， system construction and management， multi-party monitoring and intervention. Related data were selected before （Jan-Jun. 2018） and after intervention （Jul.-Dec. 2018） to evaluate intervention effects， involving paediatric outpatient/emergency intravenous infusion rate， antibiotics intravenous infusion rate， the rate of intravenous infusion prescription， total cost of antibiotics， TCM injection， adjunctive drugs and key monitoring drugs in infusion prescriptions. RESULTS： Through the comprehensive intervention of pharmacists， related indexes of outpatient/emergency intravenous infusion in paediatric were decreased greatly in our hospital. The rate of intravenous infusion， the rate of antibiotics intravenous infusion， the rate of intravenous infusion prescription and the rate of antibiotics intravenous infusion prescription were decreased from 19.52％， 15.46％， 20.29％， 11.20％ to 10.37％， 8.55％， 10.25％， 6.64％（P＜0.001）， respectively. Total cost of antibiotics， TCM injection， adjunctive drug and key monitoring drug were decreased respectively in infusion prescriptions （P＜0.001）. CONCLUSIONS： The comprehensive intervention measures taken by pharmacists in our hospital can reduce the rate of outpatient/emergency intravenous infusion in paediatric and the medical cost， and promote the safety of drug use in children.

OBJECTIVE: To explore the effects of nano-silicon dioxide( SiO_2) on the survival and poly( ADP-ribose)polymerase-1( PARP-1) expression in human bronchial epithelial cells( 16 HBE cells). METHODS: i) The 16 HBE cells were treated with nano-SiO_2 at concentrations ranging from 0 to 100 mg/L for 24. 0 hours,and CCK-8 assay was used to examine cell viability. ii) The 16 HBE cells were divided into 6 groups: solvent control group( equal volume solvent treatment),micro-SiO_2 control group( treated with 20 mg/L micro-SiO_2),5,10,and 20 mg/L nano-SiO_2 groups( treated with the corresponding final dose of nano-SiO_2),and curcumin group. The curcumin group was given pretreatment with curcumin at a final concentration of 10 μmol/L for 2. 0 hours followed by treatment with a final concentration of 20 mg/L of nano-SiO_2. Cells in each group were harvested at time points of 4. 0,12. 0 and 24. 0 hours after treatment. The relative expression of PARP-1 mRNA and protein in 16 HBE cells was detected by quantitative real-time polymerase chain reaction and Western blotting respectively. RESULTS: i) The survival of 16 HBE cells decreased with increasing nano-SiO_2 treatment dose,showing a dose-effect relationship( P < 0. 01). ii) The expression of PARP-1 mRNA and protein in 16 HBE cells were dose-dependently decreased after nano-SiO_2 stimulation at the 12. 0 and 24. 0 hours time points( P < 0. 01). The expression of PARP-1 mRNA and protein in 5,10,and 20 mg/L nano-SiO_2 groups decreased at the above mentioned time points( P < 0. 05),compared with the solvent control group at the same time points. The expression of PARP-1 mRNA and protein in 20 mg/L nano-SiO_2 group was lower than that in the micro-SiO_2 control group at the same 12. 0 and 24. 0 hours time point( P < 0. 05). The above two indexes of cells were higher in curcumin group than that of 20 mg/L nano-SiO_2 group at the 12. 0 hours time point( P < 0. 05). CONCLUSION: Nano-SiO_2 stimulation can lead to decrease survival of 16 HBE cells in a dose-dependent manner and down-regulation of PARP-1 expression may be one of the mechanisms of proliferation and inhibition of 16 HBE cells induced by nano-SiO_2. Curcumin has certain protective effect on nano-SiO_2-induced 16 HBE cell injury.

Objective To explore the influence of dexmedetomidine combined with sufentanil on postoperative analgesia effect and immune function for patients undergoing lower limb fracture surgery. Methods One hundred cases patients of lower limb fracture undergoing open reduction and internal fixation of elective surgery in our hospital from January 2016 to November 2016 were selected ( ASAⅠ-Ⅱgrade, male 51 cases and female 49 cases, 30-65 years old, 50-75 kg). All cases were randomly divided into sufentanil group (Group S) and dexmedetomidine combined with sufentanil group (Group DS) according to the random number table, who used patient-controlled intravenous analgesia (PCIA). The PCIA drug formulations of two groups were as followings: sufentanil group (Group S) used 3 μg/h sufentanil+ondansetron 16 mg, dissolved in 0.9%sodium chloride injection 100 ml;dexmedetomidine combined with sufentanil group (Group DS) used dexmedetomidine 0.1μg/(kg · h)+sufentanil 2μg/h+ondansetron 16 mg, dissolved in 0.9%sodium chloride injection 100 ml;background infusion rate was 2 ml/h, and loading dose was 2 ml. Dose for patient-controlled analgesia (PCA) was 0.5 ml, and locking time was 15 min. The changes of pain, MAP, HR , sedation scores were recorded at 4 h (T1), 8 h (T2), 24 h (T3) and 48 h (T4) after operation; nausea and vomiting, hypotension, bradycardia, respiratory depression and other adverse reactions were aslo recorded at the same time. At 10 min before induction of anesthesia (T0) and T1-T4 after operation, the CD3+, CD4+, CD8+and NK cell activity of peripheral blood T lymphocyte subsets were determined using flow cytometry, and CD4 +/CD8 + ratio was calculated. Results The levels of MAP and HR in DS group at each time point after operation were lower than those in group S (P<0.05), the level of visual analogue score (VAS) in DS group at different time points were significantly lower than those of s group: (1.8 ± 0.3) scores vs. (2.5 ± 0.5) scores, (1.1 ± 0.5) scores vs. (1.9 ± 0.3) scores, (1.0 ± 0.5) scores vs. (1.8 ± 0.5) scores, (0.8 ± 0.3) scores vs. (1.5 ± 0.6) scores (P < 0.05), Ramsay Sedation score was significantly higher than that of s group: (3.5 ± 0.3) scores vs. (2.4 ± 0.6) scores, (3.3 ± 0.5) scores vs. (2.5 ± 0.3) scores, (3.5 ± 0.6) scores vs. (2.3 ± 0.5) scores, (3.2 ± 0.4) scores vs. (2.2 ± 0.5)scores (P<0.05);nausea and vomiting occurred in two patients after the operation of group DS , which was significantly lower than that of S group (P<0.05);compared with those at T0, CD3+, CD4+, CD4+/CD8+and NK cells in the two groups of patients decreased significantly at the time of T1-T4 (P<0.05);The levels of CD3+, CD4+, CD4+/CD8+and NK cells were significantly higher in group DS at T1- T4 than those in group S(P < 0.05). Conclusions Dexmedetomidine combined with sufentanil for analgesia in patients with lower limb fracture surgery has better effect. It decreases the incidence of nausea and vomiting, and improves the cellular immune function of patients.