Practice of clinical pharmacist developing in medication reconciliation for the inpatients in cardiovascular medi- cine department / 中国药房

OBJECTIVE To analyze the medication reconciliation for the inpatients in cardiovascular medicine department ，to provide reference for the establishment of working mode of clinical pharmacists in the department of cardiovascular medicine and to provide a basis for clinical pharmacists and community pharmacists developing pharmaceutical care for patients after transfering to community health center. METHODS From October 2020 to September 2021，newly admitted or newly transferred inpatients with chronic disease were selected from Shiyan People ’s Hospital of Shenzhen Bao ’an District. Medication reconciliation was conducted by clinical pharmacists after pharmaceutical consultation. According to the Pharmaceutical Care Network Europe （PCNE） classification system V 9.1，the existing drug-related problems （DRPs）were classified and summarized. The effectiveness and safety evaluation，medication education and other measures were provided ，and the acceptance of intervention was analyzed at the same time. RESULTS A total of 100 patients were included ，including 54 males and 46 females. The average age was （60.21±9.69） years，the average number of chronic diseases was （2.84±0.83），and the median number of drugs was 5.00. Among them ，110 treatment drug deviations were found in 74 patients，involving 10 categories and 61 drugs. Top three drugs in the list of accumulative drug deviation were cardiovascular system drugs （35 deviations），digestive medicine drugs （16 deviations）and endocrine system drugs （15 deviations）. The above treatment drug deviation may cause 122 DRPs， mainly “treatment effectiveness”problems（74 DRPs），and the causes were “inappropriate medication time ormedication interval ”（32 DRPs）， followed by “inappropriate drug combination ”（10 DRPs）. Interventions to DRPs mainly concentrated on patient level ，drug level （58）and doctor level （58），155 of which （84.70％）were fully accepted and implemented. CONCLUSIONS Some patients have a weak awareness of medications according to doctor ’s advice；drug reconciliation led by clinical pharma- cists at admission can fully understand the potential drug problems of patients ，and help doctors improve the drug compliance of patients and ensure their medication safety .

Expression and characterization of recombinant wheat quiescin sulfhydryl oxidase and its effect on bread quality / 生物工程学报

Wheat quiescin sulfhydryl oxidase was expressed in Escherichia coli for developing a new biological flour improver. The synthesized wqsox gene was constructed into the vector pMAL-c5x and expressed in E. coli, then the expression conditions of recombinant protein was optimized. The MBP fusion label in recombinant protein was removed by protease digestion after affinity purification. Moreover, enzymatic properties of the purified wQSOX and its effect on bread quality were investigated. The synthesized wqsox gene contained 1 359 bp and encoded 453 amino acids with a deduced molecular weight of 51 kDa. The constructed recombinant vector pMAL-c5x-wqsox could successfully express soluble recombinant protein MBP-wQSOX in E. coli Rosetta gamiB(DE3), and the optimal induced expression conditions for recombinant protein were 25 °C, 0.3 mmol/L IPTG and 6 h. MBP fusion tag was cut out by factor Xa protease and wQSOX was prepared after affinity purification. wQSOX could catalyze the oxidation of DTT, GSH and Cys, accompanying the production of H2O2, and exhibited the highest substrate specificity for DTT. Furthermore, enzymatic properties results demonstrated that the optimal temperature and pH for wQSOX catalyzing oxidation of DTT was 50 °C and 10.0, respectively, and wQSOX presented a good stability under high temperature and alkaline environment. The addition of wQSOX with 1.1 U/g flour significantly (P<0.05) increased 26.4% specific volume of the bread, and reduced 20.5% hardness and 24.8% chewiness of bread crumb compared to the control, indicating a remarkable ability to improve the quality of bread.

Immunoglobulin fragment F(ab')2 against RBD potently neutralizes SARS-CoV-2 in vitro

COVID-19 caused by the emerging human coronavirus, SARS-CoV-2, has become a global pandemic, leading a serious threat to human health. So far, there is none vaccines or specific antiviral drugs approved for that. Therapeutic antibodies for SARS-CoV-2, was obtained from hyper immune equine plasma in this study. Herein, SARS-CoV-2 RBD with gram level were obtained through Chinese hamster ovary cells high-density fermentation. The binding of RBD to SARS-CoV-2 receptor, human ACE2, was verified and the efficacy of RBD in vivo was tested on mice and then on horses. As a result, RBD triggered high-titer neutralizing antibodies in vivo, and immunoglobulin fragment F(ab)2 was prepared from horse antisera through removing Fc. Neutralization test demonstrated that RBD-specific F(ab)2 inhibited SARS-CoV-2 with EC50 at 0.07 g/ml, showing a potent inhibitory effect on SARS-CoV-2. These results highlights as RBD-specific F(ab)2 as therapeutic candidate for SARS-CoV-2.

Cx43 silencing inhibits mechanical stress-induced apoptosis in mouse articular chondrocyte / 中南大学学报(医学版)

To explore the effect of connexin 43 (Cx43) silence on the apoptosis in mouse chondrocyte under mechanical stress.
 Methods: Mouse chondrocyte ATDC5 cells were divided into a control group, a mechanical stress group, a Cx43 siRNA transfection group, a scramble siRNA transfection group, a mechanical stress+scramble group, and a mechanical stress+siCx43 group. Flexcell FX-5000 system was used to produce mechanical stress on ATDC5 cells cultured in vitro. The mRNA and protein level of Cx43 was detected by quantitative RT-PCR (RT-qPCR) and Western blot. The cell activity and cell apoptosis was detected by cell counting kit-8 (CCK-8) method and flow cytometry, respectively. Caspase-3 activity was detected by colorimetric assay. The protein expression of Bcl-2, Bax, p-JNK and JNK was detected by Western blot.
 Results: Mechanical stress upregulated the mRNA and protein expression of Cx43 (both P<0.05). Transfection of Cx43 siRNA significantly decreased Cx43 mRNA and protein level (both P<0.05). After stimulation with mechanical stress, chondrocyte viability was significantly decreased, whereas cell apoptosis and caspase-3 activity were increased (both P<0.05). Mechanical stress obviously upregulated Bax protein level, and downregulated Bcl-2 protein expression and Bcl-2/Bax (both P<0.05). Cx43 siRNA transfection significantly increased cell viability, inhibited cell apoptosis and caspase-3 activity (both P<0.05). Cx43 siRNA also inhibited Bax expression, and increased the Bcl-2 protein expression and Bcl-2/Bax (both P<0.05). Furthermore, Cx43 siRNA significantly suppressed the p-JNK expression induced by mechanical stress (P<0.05).
 Conclusion: Cx43 silence inhibits mechanical stress-induced apoptosis in chondrocyte, which might be mediated by JNK signaling pathway.

Application of modified Veil nerve-sparing technique in laparoscopic radical prostatectomy / 中国医师进修杂志

Objective To explore the application and early efficacy of modified Veil nerve-sparing technique during laparoscopic radical prostatectomy(LRP).Methods Fifty-seven modified Veil nerve-sparing during LRP procedures were performed in patients with clinically localized prostate cancer between 2012 and 2016 by the same surgeon.Preoperative PSA level was 10.9 μg/ml,and Gleason score was 6.06(5-8).TNM clinical stage showed cT1 in 39 cases and cT2in 18 cases.All patients underwent transrectal ultrasonography before operation. Prostate volume was 40.2 (26- 99) ml. ECT bone scan excluded bone metastasis.MRI or CT examination showed no obvious prostate invasion and lymph node metastasis. The key technology was anatomical separation of detrusor apron, dorsal vascular complex (DVC) and the level between the prostate capsule, and a complete reservation was accomplished. Measurements: the rates and location of positive surgical margins (PSM) and tumor biochemical recurrence rate as well as functional outcomes were presented.Questionnaires were used to assess urine function and IIEF-5 score was used to estimate sexual function.Results Fifty-seven cases were followed up,and the average follow-up of 27.3(6-65)months.Five cases showed biochemical recurrence after 23 months.Five patients had a PSM(2 patients in apical margins,1 patient in left side,1 patient in right side and 1 patient in the bottom).At catheter removal,49 of 57 patients(86%)were dry(0 pads),and 8 of 57 patients(14%)needed one security pad.After 3 months and 6 months,42%(24 of 57 patients)and 60%(34 of 57 patients)presented an International Index of Erectile Function score>15(with or without phospho-diesterasetype-5inhibitors). Conclusions The modified Veil nerve-sparing technique during LRP can retain the fascia around the prostate more completely and restore postoperative urine and erectile function early.For selective cases, it will not increase the positive rate of surgical margins and biochemical recurrence rate.

Application of pubovesical complex preserving technique during laparoscopic radical prostatectomy / 中华泌尿外科杂志

Objective To describe a novel pubovesical complex preserving technique for laparoscopic radical prostatectomy and to evaluate its postoperative outcomes.Methods From January 2011 to May 2017,168 patients who underwent laparoscopic radical prostatectomy were enrolled and analyzed retrospectively.Their mean age were 62.8 (46-74) years,preoperative PSA 11.3ng/ml,Gleason score 6.7,preoperative prostate volume 46.5 ml.They all got preoperative potency (IIEF-5 score ≥ 15 score).TNM clinical stage:cT1 123 cases,45 cases cT2.There were 59 patients with pubovesical complex preserving technique for laparoscopic radical prostatectomy (group A):without pelvic fascia cut and deep vein complex suture.The preservation of the periprostatic anatomy was kept by preserving the pubovesical complex,including detrusor apron with pubovesical ligaments,DVC and NVB.There were 46 patients with conventional intrafascial laparoscopic radical prostatectomy (group B) and 63 patients with interfascial laparoscopic radical prostatectomy(group C).No differences were found between the three groups in terms of preoperative age,clinical staging,prostate-specific antigen (PSA) values,Gleason score at biopsy and preoperative good potency (IIEF-5 score)(P ＞0.05).Continence was defined as zero to one security pad per day.The three groups were compared for perioperative variables,PSM (positive surgical margin,PSM)rate,postoperative urinary continence functional and potency (IIEF-5 score).Biochemical recurrence-free survival was by Kaplan-Meier and log-rank.Results No differences were found in the three groups in terms of operative times,blood loss,catheterization time and postoperative stay and histologic status (PSM was similar to that of the groups (8.5％ in group A,13.0％ in group B vs.11.1％ in group C).Urinary incontinence:group A,the continence rate was 71％,82％,92％ and 100％ at 1,3 and 6 months after catheter removal,respectively;group B,the continence rate was 63％,80％,89％ and 96％ respectively;group C,it was 24％,54％,79％ and 86％ respectively.The group A showed a significantly earlier recovery from incontinence compared with that in the group C at immediately after catheter removal and 1 month after catheter remove (x2 =27.47,P ＜ 0.001;x2 =15.20,P ＜ 0.01).The group B showed a significantly earlier recovery from incontinence compared with that in the group C at immediately after catheter removal and 1 month,(x2 =17.00,P ＜ 0.01;x2 =8.20,P ＜ 0.05).No differences were found between the A and B groups at immediately after catheter removal and 1 month,(P ＞ 0.05).Regarding sexual function,at the postoperative 1,3,6 months,median IIEF-score was 10,11,16 in the group A,respectively,8,9,13 in the group B respectively,and 7,8,12 in the group C respectively.No differences were found in the three groups in potency (IIEF-5 score).Baseline IIEF-score was reached by 53％,35％ and 21％ at postoperative 6 months.There were significant differences between the A and the C groups.(x2 =13.45,P ＜0.01).There were no significant differences between the A and the B groups.(x2 =3.30,P ＞ 0.05).Follow-up was 31.6 (6-69) months.Biochemical recurrence-free survival at 3 years was 79.3％,76.3％ and 76.4％ by A,B and C group,respectively.Conclusions The pubovesical complex preserving technique for laparoscopic radical prostatectomy provides early recovery from incontinence,faster recovery of sexual function preoperative levels.

Study on the preoperative clinical criteria for nerve-sparing radical prostatectomy / 肿瘤研究与临床

Objective@#To investigate the preoperative clinical criteria for nerve-sparing radical prostatectomy.@*Methods@#A retrospective analysis of 79 patients undergoing radical prostatectomy with complete clinical and pathological data in Jinhua Hospital of Zhejiang University from January 2012 to December 2016 was performed. The distance between the edge of the prostate tumor and the neurovascular bundle (NVB) was measured. When the distance between the edge of the tumor and the ipsilateral NVB was >2 mm, NVB retention surgery can be performed; when it was ≤2 mm, NVB retention surgery cannot be performed. The influencing factors of the distance between the tumor edge and NVB were analyzed by χ 2 test and logistic regression analysis.@*Results@#Univariate analysis showed that side-specific positive biopsy core ≥1/3, side-specific maximum tumor length in biopsy core ≥5 mm, side-specific tumor involvement rate in biopsy core ≥1/2 and extraprostatic cancer extension by preoperative magnetic resonance imaging (MRI) examination were associated with the distance between the tumor edge and NVB (all P < 0.01). Multivariate analysis showed that extraprostatic cancer extension by preoperative MRI examination (OR = 3.66, P = 0.006) and side-specific positive biopsy core ≥1/3 (OR = 3.39, P = 0.008) were the independent influence factors.@*Conclusion@#The clinical criteria for a nerve-sparing radical prostatectomy are side-specific positive biopsy core <1/3 and no extraprostatic extension by preoperative MRI examination.

Enzyme Inhibitors Cause Multiple Effects on Accumulation of Monoterpene Indole Alkaloids in Catharanthus Roseus Cambial Meristematic Cell Cultures.

BACKGROUND: Enzyme inhibitors have been used for the clarification of biosynthesis of natural products. Catharanthus roseus cambial meristematic cell (CMC) culture has been established and proved to be a better monoterpeneindole alkaloid (MIA) producer than C. roseus dedifferentiated cell (DDC) culture. However, little is known about the inter-relationship of the MIA-biosynthetic genes with respect to their transcription. OBJECTIVE: To clarify effects of alteration of one gene transcription on transcript levels of another genes in MIA-biosynthetic pathway, and how the accumulation of MIAs in CMCs are influenced by the alteration of their biosynthetic gene transcript levels. MATERIALS AND METHODS: 3-Hydroxy-3-methylglutaryl-CoA reductase (HMGR) inhibitor lovastatin and 1-deoxy-D-xylulose 5-phosphate synthase (DXS) inhibitor clomazone were fed to C. roseus CMC cultures. The contents of MIAs were qualified by High Performance Liquid Chromatography and the transcript levels of the relevant genes were measured by qRT-PCR. RESULTS: Lovastatin improved the accumulation of MIAs via increasing the transcription of their biosynthetic genes encoding DXS1, tryptonphan decarboxylase (TDC), loganic acid methyltransferase (LAMT), strictosidine synthase (STR), desacetoxyvindoline-4-hydroxylase (D4H) and ORCA3 (a jasmonate-responsive transcriptional regulator), whereas clomazone reduced the contents of MIAs and the mRNA levels of the corresponding genes. CONCLUSION: The biosynthesis of MIAs in C. roseus is is manipulated via a complex mechanism, the knowledge of which paves the way for rationally tuning metabolic flux to improve MIA production in C. roseus CMCs.

The healing of alveolar bone defects with novel bio-implants composed of Ad-BMP9-transfected rDFCs and CHA scaffolds.

Cells, scaffolds, and growth factors play important roles in bone regeneration. Bone morphogenetic protein 9 (BMP9), a member of BMP family, could facilitate osteogenesis by regulating growth factors and promoting angiogenesis. Similar to other stem cells, rat dental follicle stem cells (rDFCs), the precursor cells of cementoblasts, osteoblasts and periodontal ligament cells, can self-renew and exhibit multipotential capacity. Coralline hydroxyapatite (CHA) has good biocompatibility and conductivity required for bone tissue engineering. Here, we reported that BMP9 could enhance the osteogenic differentiation of rDFCs in cell culture. Moreover, our results suggested that BMP9 acted through the Smad1/5/8 signaling pathway. We also produced a novel scaffold that encompasses bio-degradable CHA seeded with recombinant adenoviruses expressing BMP9-transfected rDFCs (Ad-BMP9-transfected rDFCs). With this implant, we achieved more alveolar bone regeneration in the alveolar bone defect compared to blank group, CHA group and rDFCs group. Our results provided a novel bio-implants composed of Ad-BMP9-transfected rDFCs and CHA scaffolds and its mechanism is regarding the activation of Smad1/5/8 signaling pathway in BMP9-induced rDFCs osteogenesis.

BMP9 effectively induces osteogenic differentiation of immortalized calvarial mesen-chymal progenitor cells / 军事医学

Objective To investigate the effect of adenovirus-bone morphogenic protein 9 ( Ad-BMP9 ) on osteogenic differentiation of immortalized calvarial mesenchymal progenitor cells ( iCALs ) .Methods iCALs were infected with adenoviral vectors encoding BMP-9 or green fluorescent protein ( GFP) and the early osteogenic differentiation was assessed by detecting alkaline phosphatase (ALP) activity after being cultured for 3, 5 and 7 days.14 days after infection, alizarin red S staining was performed to study the formation of osteogenic calcium nodules .The expression of osteogenic marker genes Runx2 and OCN was assessed by quantitative real-time ( RT )-PCR and Western blotting .Results Significant increases in ALP activity and in the expressions of Runx 2 and OCN were detected in BMP-9 treated iCALs compared with GFP-treated cells(P<0.05).Meanwhile, alizarin red S staining showed that more mineralized nodules were found in the BMP-9 induced group .Conclusion BMP-9 can promote the osteogenic differentiation of iCALs .

Establishment of reproducible xenotransplantation model of T cell acute lymphoblastic leukemia in NOD/SCID mice / 华中科技大学学报（医学）（英德文版）

T cell acute lymphoblastic leukemia (T-ALL) is an aggressive leukemia. However the poor prognosis and low morbidity restrict further analysis of the disease. Therefore there is an increasing demand to develop animal models for identifying novel therapeutic approaches. In this study, we inoculated the anti-mouse CD122 monoclonal antibody conditioned NOD/SCID mice with the leukemia cells from 9 T-ALL patients and 1 cell line via the tail vein. Four of the 9 patients and the cell line were successfully engrafted. Flow cytometry detected high percentage of human CD45(+) cells in recipient mice. Immunohistochemistry showed infiltration of human CD45(+) cells in different organs. Serial transplantation was also achieved. In vivo drug treatment showed that dexamethasone could extend survival, which was consistent with clinical observation. These results demonstrated that we successfully established 5 xenotransplantation models of T-ALL in anti-mCD122 mAb conditioned NOD/SCID mice, which recapitulated the characteristics of original disease.

Establishment of reproducible xenotransplantation model of T cell acute lymphoblastic leukemia in NOD/SCID mice / 华中科技大学学报（医学）（英德文版）

T cell acute lymphoblastic leukemia (T-ALL) is an aggressive leukemia. However the poor prognosis and low morbidity restrict further analysis of the disease. Therefore there is an increasing demand to develop animal models for identifying novel therapeutic approaches. In this study, we inoculated the anti-mouse CD122 monoclonal antibody conditioned NOD/SCID mice with the leukemia cells from 9 T-ALL patients and 1 cell line via the tail vein. Four of the 9 patients and the cell line were successfully engrafted. Flow cytometry detected high percentage of human CD45(+) cells in recipient mice. Immunohistochemistry showed infiltration of human CD45(+) cells in different organs. Serial transplantation was also achieved. In vivo drug treatment showed that dexamethasone could extend survival, which was consistent with clinical observation. These results demonstrated that we successfully established 5 xenotransplantation models of T-ALL in anti-mCD122 mAb conditioned NOD/SCID mice, which recapitulated the characteristics of original disease.