Guided endodontics for the treatment and vital pulp concervation of type Ⅲ Dens Invaginatus in an immature maxillary lateral incisor with periapical periodontitis / 实用口腔医学杂志

A young maxillary lateral incisor of Oehlers type Ⅲ Dens invaginatus with peri-invagination periodontitis was reconstructed by CBCT,with the help of guided endodontics,the pathway to invagination was successfully established.The invaginated pseudo-root canal was treated with Vitapex mediation while preserving the pulp.After 6-month follow-up,the tooth was clinically asymptomatic.Radiological ex-amination indicated complete healing of the peri-invagination lesion with narrowed open apex,and the thickened root canal wall.

Construction of a risk prediction model of lung involvement based on chest CT and clinical features in patients with primary Sjogren's syndrome / 实用医学杂志

Objective To construct a risk prediction model of pulmonary involvement based on chest CT and clinical feature in patients with primary Sjogren's syndrome(pSS),and to explore the risk prediction value of the model.Methods A total of 360 pSS patients who had been treated at Handan Hospital of Traditional Chinese Medicine from October 2020 to August 2023 were retrospectively selected as study objects,and were then divided into a modeling group(252 patients)and a verification group(108 patients)according to a ratio of 7∶3.The patients in the modeling group were divided into a control group(201 patients)and an involvement group(51 patients)based on presence or absence of lung involvement.The data on clinical characteristics and features of chest high-resolution CT(HRCT)in the modeling group was collected.Univariate analysis was performed among the groups to determine the relevant factors affecting lung involvement in pSS patients.Binary logistic regression analysis was performed on related factors to screen independent risk factors.A prediction model was established based on the independent risk factors.A verification and value analysis of the column-line prediction model were completed through data collection of the verification group.Results Age,disease course,cough,Raynaud's phenomenon,C-reactive protein(CRP),anti-SSA antibody,and HRCT were the relevant factors affecting lung involvement in pSS patients(all P＜0.05).Further binary logistic regression analysis showed that old age,prolonged disease course,cough and abnormal HRCT imaging were independent risk factors for lung involvement in SS patients(all P＜0.05).A nomogram risk prediction model was constructed based on independent factors.The model verification results indicated that the calibration chart showed better performance in the prediction model.The AUC of the area under the receiver operating characteristic(ROC)curve was 0.993 the modeling group and 0.995 in the validation group.Conclusions The clinical characteristics and the results of chest CT are closely related with lung involvement in patients with pSS.Old age,prolonged disease course,cough,and abnormal HRCT imaging are independent risk factors affecting lung involvement in patients with pSS.The prediction model established on this basis has a higher predictive value for the occurrence of lung involvement in patients receiving after-loading radiotherapy.

Expression of Trichinella spiralis trehalase(TsTRE)protein and establishment of an indirect ELISA / 中国人兽共患病学报

We established an indirect ELISA method using Trichinella spiralis trehalase(TsTRE)protein expressed in prokaryotic cells.The TsTRE gene was amplified by RT-PCR and ligated into the pCold I plasmid,which was expressed in E.coli BL21 competent cells.The rTsTRE protein was purified through affinity column chromatography.The TsTRE protein was localized with immunofluorescence techniques,and the immunogenicity of rTsTRE was detected by westernblotting.Subse-quently,rTsTRE protein was used as a coating antigen to establish an indirect ELISA.We optimized the antigen-coating con-centration,serum dilution concentration,antigen-coating incubation time,type of blocking solution,blocking incubation time,HRP-labeled goat anti-rabbit IgG serum dilution concentration,HRP-labeled goat anti-rabbit IgG serum incubation time and response time of TMB.Subsequently,the critical value,repeatability,sensitivity,specificity and clinical detection rate of the ELISA were evaluated.Immunofluorescence indicated that trehalase was abundant in the rod-shaped body,tail and epidermis of Trichinella spiralis muscle larvae.Western-blot indicated that rTsTRE protein combined with the positive serum of mice infected with T.spiralis for 42 d;the band was approximately 60 kDa.The established indirect ELISA had a positive threshold of 0.384;the intra-run and inter-run coefficients of variation were 5.504％-7.630％ and 4.664％-9.929％,and did not exceed 10％.The lowest detectable titer was 1:1 280.No cross reaction was observed with antibodies to Clonorchissinensis,Schistosoma ja-ponicum,Ascaris suum,Toxocara gondii and Toxocara canis,and the clinical negative detection rate was 0％.Thus,we suc-cessfully expressed the rTsTRE protein.Moreover,the established indirect ELISA method using the TsTRE protein as the coating antigen had good repeatability,sensitivity,specificity and clinical detectability,and can be applied to the detection of clinical samples.

Evaluation of scientific research efficiency based on Data Envelopment Analysis in a third-class hospital in Xinjiang / 中华医学科研管理杂志

Objective:To understand the overall development and dynamic change trend of scientific research input-output efficiency of a tertiary hospital in Xinjiang from 2016 to 2021, so as to provide reference and basis for optimizing hospital scientific research configuration and improving scientific research efficiency.Methods:The BCC model of Data Envelopment Analysis (DEA) method is combined with Malmquist index method for analysis and evaluation.Results:Within 6 years, the average value of comprehensive technical efficiency of nursing units has not reached 1.The total factor productivity of nursing units was unstable, showing an alternating rise and fall trend, with a fluctuation range of 0.749~1.140.Conclusions:The overall efficiency of scientific research in clinical departments of hospitals is not high. Hospitals should implement multi-level scientific research evaluation, form a dynamic evaluation system for scientific research, reasonably optimize and supervise the investment and utilization of scientific research resources, improve the scientific research management system, enhance the scientific research awareness of researchers, promote technological progress of researchers, and thereby improve scientific research efficiency.

Analysis of the current situation and factors influencing social isolation of maintenance hemodialysis patients / 中国实用护理杂志

Objective:To investigate the current situation of occurrence of social isolation in maintenance hemodialysis patients and analyze its influencing factors, in order to provide a theoretical basis for the implementation of targeted interventions.Methods:The study was a cross-sectional study. A total of 315 dialysis patients in three hemodialysis centers in Xinjiang were selected by simple sampling method from October 2021 to March 2022, who were investigated using the general information questionnaire, Lubben-6 social network scale, International Physical Activity Questionnaire (sedentary behavior section), Patient Health Questionnaire 9-item scale (PHQ-9) and Frail Scale. Univariate analysis and binary Logistic regression analysis were used to explore the influencing factors affecting the social isolation of dialysis patients.Results:Social isolation was present in 28.89% (91/315) of maintenance hemodialysis patients, and binary Logistic regression analysis revealed that age (45-59 years old: OR=4.815, 95% CI 1.362-17.017;≥60 years old: OR=8.968, 95% CI 2.349-34.236), dialysis age ( OR=2.788, 95% CI 1.334-5.826), sedentary behavior ( OR=2.504, 95% CI 1.406-4.461), depression ( OR=2.095, 95% CI 1.179-3.722), and debilitation ( OR=2.043, 95% CI=1.062-3.933) were influencing factors of social isolation in maintenance hemodialysis patients (all P<0.05). Conclusions:The occurrence of social isolation in maintenance hemodialysis patients was closely associated with advanced age, high dialysis age, sedentary behavior, debilitation, and depressive status, suggesting that medical staff in hemodialysis centers can implement targeted interventions to prevent or improve the level of social isolation in patients based on relevant influencing factors.

Alcohol extract of root and root bark of Toddalia asiatica alleviates CIA in rats through anti-inflammatory and proapoptotic effects / 中国中药杂志

This study aims to investigate the therapeutic effect of alcohol extract of root and root bark of Toddalia asiatica(TAAE) on collagen-induced arthritis(CIA) in rats through phosphatidylinoinosidine-3 kinase/protein kinase B(PI3K/Akt) signaling pathway. To be specific, CIA was induced in rats, and then the rats were treated(oral, daily) with TAAE and Tripterygium Glycoside Tablets(TGT), respectively. The swelling degree of the hind leg joints was scored weekly. After 35 days of administration, the histopathological changes were observed based on hematoxylin and eosin(HE) staining. Enzyme-linked immunosorbent assay(ELISA) was employed to detect the levels of cytokines [tumor necrosis factor-α(TNF-α), interleukin(IL)-6)]. Terminal deoxynucleotidyl transferase dUTP nick end labeling(TUNEL) staining was performed to detect the apoptosis of synoviocytes in rats. Western blot was used to detect the expression levels of apoptosis-related proteins B-cell lymphoma 2(Bcl-2)-associated X(Bax), Bcl-2, and caspase-3 and pathway-related proteins phosphoinositide 3-kinase(PI3K), phosphorylated(p)-PI3K, protein kinase B(Akt), and p-Akt. RT-qPCR was conducted to examine the mRNA levels of Bax, Bcl-2, caspase-3, TNF-α, IL-6, and IL-1β and pathway-related proteins PI3K, p-PI3K, Akt, and p-Akt. TAAE can alleviate the joint swelling in CIA rats, reduce serum levels of inflammatory cytokines, improve synovial histopathological changes, promote apoptosis of synoviocytes, and inhibit synovial inflammation. In addition, RT-qPCR and Western blot results showed that TAAE up-regulated the level of Bax, down-regulated the level of Bcl-2, and activated caspase-3 to promote apoptosis in synoviocytes. TAAE effectively down-regulated the protein levels of p-PI3K and p-Akt. In this study, TAAE shows therapeutic effect on CIA in rats and reduces the inflammation. The mechanism is that it suppresses PI3K/Akt signaling pathway and promotes synoviocyte apoptosis. Overall, this study provides a new clue for the research on the anti-inflammatory mechanism of TAAE and lays a theoretical basis for the better clinical application of TAAE in the treatment of inflammatory and autoimmune diseases.

Establishment the Evaluation System of Scientific Research Performance for Medical Staff in General Hos-pital / 中国医院管理

Objective Establishing an integrated assessment index system in order to strengthen the multi-dimen-sional evaluation for scientific research performance of medical staff in tertiary general hospital.Methods The method of documentary and Delphi are used in filtering the indexes of medical personnel's scientific research performance evaluation and the method of analytic hierarchy process is used in ascribing different weights for each index that the consistency is tested.Results There are two evaluation index systems in the Evaluation System of medical research performance in tertiary general hospital.The first one is Medical Staff's Scientific Research Ability Evaluation System.There are 51 indexes in the system,including 2 first-level indexes,1 second-level indexes and 38 third-level indexes.The second system is Scientific Research Performance Evaluation Index System,which have 70 indicators,including 4 first-level indicators,15 second-level indicators,51 third-level indicators.Conclusion The index system of scientific research performance evaluation has a high credibility,and can be popularized as an index of scientific research performance evaluation for medical staff in hospital based on the practice evaluation.The system could provide data support for hospital to implement more accurate scientific research management policy.

Clinical management and prognosis for descending necrotizing mediastinitis / 中华耳鼻咽喉头颈外科杂志

Objective: To investigate the clinical characteristics, treatment experiences and prognostic factors for descending necrotizing mediastinitis (DNM). Methods: A retrospective analysis was performed on the data of 22 patients with DNM diagnosed and treated in Henan Provincial People's Hospital from January 2016 to August 2022, including 16 males and 6 females, aged 29-79 years. After admission, all patients underwent CT scanning of the maxillofacial, cervical, and thoracic regions to confirm their diagnoses. Emergency incision and drainage were performed. The neck incision was treated with continuous vacuum sealing drainage. According to the prognoses, the patients were divided into cure group and death group, and the prognostic factors were analyzed. SPSS 25.0 software was used to analyze the clinical data. Rusults: The main complaints were dysphagia (45.5%, 10/22) and dyspnea (50.0%, 11/22). Odontogenic infection accounted for 45.5% (10/22) and oropharyngeal infection accounted for 54.5% (12/22). There were 16 cases in the cured group and 6 cases in the death group, with a total mortality rate of 27.3%. The mortality rates of DNM typeⅠand typeⅡwere respectively 16.7% and 40%. Compared with the cured group, the death group had higher incidences for diabetes, coronary heart disease and septic shock (all P<0.05). There were statistically significant differences between the cure group and the death group in procalcitonin level (50.43 (137.64) ng/ml vs 2.92 (6.33) ng/ml, M(IQR), Z=3.023, P<0.05) and acute physiology and chronic health evaluation Ⅱ(APACHEⅡ) score (16.10±2.40 vs 6.75±3.19, t=6.524, P<0.05). Conclution: DNM is rare, with high mortality, high incidence of septic shock, and the increased procalcitonin level and APACHE Ⅱ score combined diabetes and coronary heart disease are the poor prognostic factors for DNM. Early incision and drainage combined with continuous vacuum sealing drainage technique is a better way to treat DNM.

Efficacy and safety of mitoxantrone hydrochloride liposome injection in treatment of peripheral T-cell lymphomas: a multicenter, non-interventional, ambispective cohort, real-world study (MOMENT) / 白血病·淋巴瘤

Objective:To evaluate the efficacy and safety of mitoxantrone hydrochloride liposome injection in the treatment of peripheral T-cell lymphoma (PTCL) in a real-world setting.Methods:This was a real-world ambispective cohort study (MOMENT study) (Chinese clinical trial registry number: ChiCTR2200062067). Clinical data were collected from 198 patients who received mitoxantrone hydrochloride liposome injection as monotherapy or combination therapy at 37 hospitals from January 2022 to January 2023, including 166 patients in the retrospective cohort and 32 patients in the prospective cohort; 10 patients in the treatment-na?ve group and 188 patients in the relapsed/refractory group. Clinical characteristics, efficacy and adverse events were summarized, and the overall survival (OS) and progression-free survival (PFS) were analyzed.Results:All 198 patients were treated with mitoxantrone hydrochloride liposome injection for a median of 3 cycles (range 1-7 cycles); 28 cases were treated with mitoxantrone hydrochloride liposome injection as monotherapy, and 170 cases were treated with the combination regimen. Among 188 relapsed/refractory patients, 45 cases (23.9%) were in complete remission (CR), 82 cases (43.6%) were in partial remission (PR), and 28 cases (14.9%) were in disease stabilization (SD), and 33 cases (17.6%) were in disease progression (PD), with an objective remission rate (ORR) of 67.6% (127/188). Among 10 treatment-na?ve patients, 4 cases (40.0%) were in CR, 5 cases (50.0%) were in PR, and 1 case (10.0%) was in PD, with an ORR of 90.0% (9/10). The median follow-up time was 2.9 months (95% CI 2.4-3.7 months), and the median PFS and OS of patients in relapsed/refractory and treatment-na?ve groups were not reached. In relapsed/refractory patients, the difference in ORR between patients with different number of treatment lines of mitoxantrone hydrochloride liposome injection [ORR of the second-line, the third-line and ≥the forth-line treatment was 74.4% (67/90), 73.9% (34/46) and 50.0% (26/52)] was statistically significant ( P = 0.008). Of the 198 PTCL patients, 182 cases (91.9%) experienced at least 1 time of treatment-related adverse events, and the incidence rate of ≥grade 3 adverse events was 66.7% (132/198), which was mainly characterized by hematologic adverse events. The ≥ grade 3 hematologic adverse events mainly included decreased lymphocyte count, decreased neutrophil count, decreased white blood cell count, and anemia; non-hematologic adverse events were mostly grade 1-2, mainly including pigmentation disorders and upper respiratory tract infection. Conclusions:The use of mitoxantrone hydrochloride liposome injection-containing regimen in the treatment of PTCL has definite efficacy and is well tolerated, and it is a new therapeutic option for PTCL patients.

Efficacy of the herbal pair, Radix Achyranthis Bidentatae and Eucommiae Cortex, in preventing glucocorticoid-induced osteoporosis in the zebrafish model / 中西医结合学报

OBJECTIVE@#In traditional Chinese medicine, the herbal pair, Radix Achyranthis Bidentatae (RAB) and Eucommiae Cortex (EC), is widely used to treat osteoporosis. Herein, we determined whether this herbal pair can be used to ameliorate glucocorticoid (GC)-induced osteoporosis (GIOP) and find its optimal dosage in zebrafish.@*METHODS@#The characteristics of the aqueous extract of RAB and EC were separately characterized using high-performance liquid chromatography. Osteoporosis was induced in 5-day post-fertilization zebrafish larvae by exposing them to 10 μmol/L dexamethasone (Dex) for 96 h. Seven combinations of different ratios of RAB and EC were co-administered. Treatment efficacy was determined by calculating zebrafish vertebral area and sum brightness, via alizarin red staining, and by detecting alkaline phosphatase (ALP) activity. Multiple regression analysis was conducted to test the optimal dosage ratio.@*RESULTS@#According to the Chinese Pharmacopoeia (2015), β-ecdysone (β-Ecd) is a major bioactive marker in RAB extract, while pinoresinol diglucoside (PDG) is the major marker in EC extract. Both of β-Ecd and PDG content values aligned with the Chinese Pharmacopoeia standards. Treatment with 10 μmol/L Dex reduced zebrafish vertebral area, sum brightness, and ALP activity, but RAB and EC attenuated these effects. Combining 50 µg/mL RAB and 50 µg/mL EC was optimal for preventing GIOP in zebrafish. Reverse transcription-quantitative polymerase chain reaction was used to evaluate the mRNA expression of osteogenesis-related genes. A treatment of 10 μmol/L Dex decreased runt-related transcription factor 2 (Runx2), osteogenic protein-1 (OP-1), bone γ-carboxyglutamic acid-containing protein (BGLAP), and β-catenin levels. This effect was counteracted by RAB and EC co-treatment (P < 0.05). Additionally, the effect of using the two herbal extracts together was better than single-herb treatments separately. These results demonstrated that RAB and EC preserve osteoblast function in the presence of GC. The best mass ratio was 1:1.@*CONCLUSION@#RAB and EC herbal pair could ameliorate GC-induced effects in zebrafish, with 1:1 as the optimal dosage ratio.

Effect of Shenmai Injection on Long-Term Prognosis of Patients with Chronic Heart Failure: A Multicenter, Large Sample Capacity, Long-Term Follow-Up Retrospective Cohort Study / 中国结合医学杂志

OBJECTIVE@#To explore the effect of Shenmai Injection (SMI) on the long-term prognosis of patients with chronic heart failure (CHF).@*METHODS@#The Hospital Information System was used to extract data of CHF patients, and the retrospective cohort study was conducted for analysis. In non-exposed group, standardized Western medicine treatment and Chinese patent medicine or decoction were applied without combination of SMI while in the exposed group, SMI were applied for more than 7 days. Evaluation indicators are followed with New York Heart Association functional classification (NYHA classification), left ventricular ejection fraction (LVEF), N-terminal brain natriuretic peptide precursor (NT-ProBNP), cardiogenic death and heart failure (HF) readmission. Statistical analysis includes Kaplan-Meier analysis and Cox regression which are used to explore the relationship between SMI and outcome events.@*RESULTS@#A total of 1,211 eligible CHF patients were involved and finally 1,047 patients were followed up successfully. After treatment, the cases of NYHA classification decline in the exposed and non-exposed groups accounted for 64.30% and 43.45%, respectively; the improvement values of LVEF were 8.89% and 7.91%, respectively; the improvement values of NT-ProBNP were 909 pg/mL and 735 pg/mL, respectively. After exposure on SMI, the rates of cardiogenic death and HF readmission reduced from 15.43% to 10.18% and 38.93% to 32.37%. According to Kaplan-Meier analysis, the log-rank P value of SMI and cardiogenic death was 0.014, while the counterpart of SMI and HF readmission was 0.025. Cox regression analysis indicated that for cardiogenic death, age, cardiomyopathy, diabetes, and NYHA classification were risk factors while β-blockers, aldosterone receptor antagonists, Chinese patent medicine/decoction and SMI were protective factors. Likewise, for HF readmission, age, cardiomyopathy, and NYHA classification were risk factors while SMI was a protective factor.@*CONCLUSION@#Combination with SMI on the standardized Western medicine treatment can effectively reduce cardiogenic mortality and readmission rate in CHF patients, and thereby improve the long-term prognosis.

Research on the application of CDIO teaching mode in the cultivation of core competence of PICC specialist nurses / 中国实用护理杂志

Objective:To explore the application effect of CDIO (Conceire, Design, Implement, Operate) teaching mode in the training of PICC nurse trainees.Methods:A total of 80 PICC nurses trained in Xinjiang Uygur Autonomous Region People ′s Hospital from March to August 2021 were divided into the control group (40 patients) and the intervention group (40 patients) according to the baseline comparison principle. The control group participated in the first training using the traditional teaching mode, and the intervention group participated in the second training using the CDIO teaching mode. The scores of core competence and teaching satisfaction of PICC specialist nurses in the two groups were compared at the 8th week after training, and the theoretical and operational scores of the two groups were compared at the 4th and 8th week after training. Results:After training, the scores of professional theoretical knowledge, professional operation skills and related abilities in the core competence of PICC specialist nurses in the intervention group were (27.76 ± 2.07), (40.42 ± 4.12) and (11.42 ± 1.86), respectively. The control group was (22.21 ± 3.71), (34.84 ± 6.86) and (10.15 ± 2.10) points, respectively, and the differences between the two groups were statistically significant ( t=-8.25, -4.42, -2.85, all P<0.01). After 4 and 8 weeks of training, the theoretical scores of the intervention group were (82.00 ± 7.51), (86.25 ± 3.53) points, and the operational scores of PICC laying and maintenance were (93.68 ± 3.25), (91.13 ± 4.75) points, respectively. The control group was (76.48 ± 9.38), (83.28 ± 4.05), (91.48 ± 3.72), (88.58 ± 6.28) points, respectively, and the differences between the two groups were statistically significant ( t values were -3.50 - -2.05, all P<0.05). In the teaching satisfaction of the intervention group, the teaching level, teaching method and teaching effect were 97.5% (39/40), 95.0% (38/40), 100.0% (40/40), while that of control group were 77.5% (31/40), 80.0% (32/40), 85.0% (34/40). There were statistically significant differences between the two groups ( χ2=7.31, 4.11, 4.51, all P<0.05). Conclusions:CDIO teaching mode is superior to traditional teaching mode. It can improve students′ core competence and achievement of theoretical and operational knowledge, and improve teaching satisfaction. It is worth promoting.

Prevalence and Risk Factors of Postdialysis Fatigue in Patients Under Maintenance Hemodialysis: A Systematic Review and Meta-Analysis

Purpose@#Despite the high prevalence of postdialysis fatigue (PDF) in maintenance hemodialysis patients, no meta-analysis on the prevalence and risk factors of PDF has yet been published. This study aimed to identify the prevalence of PDF and explore its related factors. @*Methods@#PubMed, Embase, CENTRAL, Web of Science, Cumulative Index to Nursing and Allied Health Literature (CINAHL), and the four Chinese databases (National Knowledge Infrastructure [CNKI], Chinese Biomedical Literature database [SinoMed], Wanfang Digital Periodicals [WANFANG], and Chinese Science and Technology Periodicals [VIP] database) were searched from inception up to July 2022. This study was performed according to the Preferred Reporting Items for Systematic Reviews and Meta-Analysis guidelines. The articles were independently searched by two reviewers, and the relevant data were extracted. The Agency for Healthcare Research and Quality was used to assess the quality of the included studies. @*Results@#Thirteen articles with 2,118 participants were included. The pooled prevalence was 60.0%. The meta-analysis results revealed that the ultrafiltration volume, mean arterial pressure after dialysis, and good sleep quality were potentially associated with PDF, whereas only good sleep quality (odds ratio 0.24, 95% confidence interval 0.19e0.30) was significantly associated with PDF. @*Conclusion@#PDF is common in maintenance hemodialysis patients, which is related to the ultrafiltration volume, sleep quality, and mean arterial pressure after dialysis. However, the mechanism underlying the risk factors and PDF remains unknown. Further research is warranted to investigate the risk factors, intervention, treatment, and mechanism in maintenance hemodialysis patients.

Cannabidiol prevents depressive-like behaviors through the modulation of neural stem cell differentiation / 医学前沿

Chronic stress impairs radial neural stem cell (rNSC) differentiation and adult hippocampal neurogenesis (AHN), whereas promoting AHN can increase stress resilience against depression. Therefore, investigating the mechanism of neural differentiation and AHN is of great importance for developing antidepressant drugs. The nonpsychoactive phytocannabinoid cannabidiol (CBD) has been shown to be effective against depression. However, whether CBD can modulate rNSC differentiation and hippocampal neurogenesis is unknown. Here, by using the chronic restraint stress (CRS) mouse model, we showed that hippocampal rNSCs mostly differentiated into astrocytes under stress conditions. Moreover, transcriptome analysis revealed that the FoxO signaling pathway was involved in the regulation of this process. The administration of CBD rescued depressive-like symptoms in CRS mice and prevented rNSCs overactivation and differentiation into astrocyte, which was partly mediated by the modulation of the FoxO signaling pathway. These results revealed a previously unknown neural mechanism for neural differentiation and AHN in depression and provided mechanistic insights into the antidepressive effects of CBD.

Effect of icaritin on proliferation and apoptosis of human ovarian cancer cells SKOV3 / 中国中药杂志

Based on the PI3K/Akt signaling pathway, this study aimed to observe the proliferation and apoptosis of ovarian cancer SKOV3 cells at different concentrations of icaritin, in order to explore the possible molecular mechanisms. The research object was ovarian cancer SKOV3 cells. The cells were divided into the control group and icaritin groups(5, 10, 20 μmol·L~(-1)), and administrated with drugs for 48 hours. The cell counting kit-8(CCK-8)assay was used to detect the inhibitory effect of icaritin on the proliferation of ovarian cancer SKOV3 cells. The proliferation ability of the SKOV3 cells was detected by EdU assay. Hoechst 33342 fluorescence staining was used to observe the apoptotic morphology of SKOV3 cells in each group. The distribution of cell cycle and the apoptosis rate of each group were detected by flow cytometry. Quantitative Real-time PCR was used to detect mRNA expressions of PTEN, PI3K, Akt in each group of cells. Protein expressions of PTEN, PI3K, Akt and p-Akt were measured by Western blot. The results showed that the cell inhibition rates of icaritin groups were significantly increased compared with the control group(P<0.05). The rates of EdU-positive cells of icaritin groups were significantly decreased(P<0.05). SKOV3 cells in icaritin groups showed morphological changes of apoptosis. Apoptosis rates of icaritin groups were significantly increased(P<0.05). The proportions of cells in G_0/G_1 phase of icaritin groups were decreased(P<0.05), while the proportions of S phase cells were increased(P<0.05). The gene and protein expressions of PTEN in icaritin groups were elevated(P<0.05). The gene expressions of PI3K and Akt in icaritin groups were down-regulated(P<0.05). The protein expression of PI3K and p-Akt in icaritin groups were reduced(P<0.05). These results indicated that icarin may inhibit the proliferation of ovarian cancer SKOV3 cells in vitro, induce cell apoptosis and affect the cycle distribution of cells by inhibiting the PI3K/Akt signaling pathway.

Effect of Icaritin on Proliferation， Apoptosis， Migration and Invasion of Human Ovarian Cancer A2780 Cells / 中国实验方剂学杂志

Objective:To study the effect of icaritin on the proliferation， apoptosis， migration and invasion of human epithelial ovarian cancer A2780 cells and the inhibitory mechanism of icaritin against cell invasion and migration via the regulation of epithelial-mesenchymal transformation （EMT）-related molecule expression. Method:A2780 cells were divided into the blank control group and low-， medium-， and high-dose （5， 10， 20 μmol·L^-1） icaritin groups and received the corresponding inventions for 48 h. Cell proliferation and viability were detected using the cell counting kit-8 （CCK-8）. The cellular proliferation inhibition and apoptosis rates were assayed by flow cytometry. The cell invasion and migration were observed in Scratch test and transwell test， followed by the calculation of wound healing rate and migration rate. The protein and mRNA expression levels of EMT-related molecules including E-cadherin， N-cadherin， and Vimentin and tumor invasion and migration-related molecule matrix metalloproteinase-9 （MMP-9） were measured by Western blot and real-time polymerase chain reaction （Real-time PCR）. Result:As revealed by CCK-8 assay and flow cytometry， compared with the blank control group， the icaritin groups all exhibited elevated proliferation inhibition rate （<italic>P</italic><0.01） and apoptosis rate （<italic>P</italic><0.05）. According to the Scratch test and transwell test， compared with the blank control group， the icaritin groups displayed weakened invasion and migration ability and decreased number and rate of cell invasion and migration （<italic>P</italic><0.05）. Western blot and Real-time PCR results showed that the protein and mRNA expression levels of N-cadherin， MMP-9 and Vimentin in each icaritin group were down-regulated as compared with those in the blank control group， while the expression of E-cadherin was up-regulated. Conclusion:Icaritin inhibits the proliferation and promotes the apoptosis of human ovarian cancer A2780 cells， and it inhibits the invasion and migration of A2780 cells possibly by regulating the expression of EMT-related molecules.

Effect of Fushengong Prescreption on Regulation-antagonism Effect of ACE-AngⅡ-AT1R Axis and ACE2-Ang(1-7)-MASR Axis of Rats with Chronic Renal Failure / 中国实验方剂学杂志

Objective:To study the effect of Fushengong prescreption on the regulation-antagonism effect of angiotensin converting enzyme-angiotensin Ⅱ-angiotensin Ⅱ 1 receptor （ACE-AngⅡ-AT1R） axis and angiotensin converting enzyme 2-angiotensin （1-7）-Mas receptor［ACE2-Ang（1-7）-MASR］ axis of rats with chronic renal failure（CRF）， and to explore its mechanism of delaying the development of CRF. Method:The 65 male SD rats were randomly divided into normal group （<italic>n</italic>=10） and modeling group （<italic>n</italic>=55）. The normal group was routinely reared， while the modeling group were administered by gavage with 0.25 g·kg^-1d^-1adenine suspension for 28 days. After the model was successfully established， the survival model rats were randomly divided into model group， benazepril group（0.01 g·kg^-1·d^-1）and low，medium and high dose of Fushengong prescreption groups （4，8，16 g·kg^-1·d^-1）. The normal group and model group were administered the same volume of normal saline by gavage， lasted for 28 days. After the experiment， systolic blood pressure （SBP） and diastolic blood pressure （DBP） of caudal artery were measured， and 24-hour urine was collected to determine 24-hour urine protein （24 h U-pro）. The content of serum creatinine（SCr） and blood urea nitrogen （BUN） in the serum were measured， the histological morphology was observed by hematoxylin eosin（HE）staining， and the degree of renal interstitial fibrosis was observed by Masson staining. Enzyme linked immunosorbent assay （ELISA） was used to determine the contents of AngⅡ， Ang （1-7） and Cystatin C （CysC） in serum and renal homogenate. The protein level of ACE， ACE2， AT1R and MASR were detected by Western blot. The expression of ACE and ACE2 protein in renal tissues were detected by immunohistochemistry. Result:Compared with normal group， the expression levels of SCr， BUN and CysC in model group were significantly increased（<italic>P</italic><0.05）， the content of AngⅡ in serum and kidney tissues were significantly increased， the content of Ang （1-7） were significantly decreased（<italic>P</italic><0.05）， the expression of ACE and AT1R protein in renal tissues were significantly increased（<italic>P</italic><0.05）， and the expression of ACE2 and MASR protein were significantly decreased（<italic>P</italic><0.05）. Compared with model group and benazepril group， after the intervention with Fushengong prescreption， the serum SCr，BUN and CysC decreased（<italic>P</italic><0.05），the content of AngⅡ in serum and kidney tissues decreased significantly，Ang（1-7） increased significantly（<italic>P</italic><0.05）， the expression of ACE and AT1R protein in renal tissues decreased significantly（<italic>P</italic><0.05）， ACE2 and MASR protein increased significantly（<italic>P</italic><0.05）. The high-dose Fushengong prescreption has the best effect. The high， medium and low-dose effects of Fushengong prescreption were dose-dependent. Conclusion:Fushengong prescreption improved renal function and pathological change of kidney in adenine-induced rats with chronic renal failure. The mechanism may be related to the inhibition of ACE-AngⅡ-AT1R axis and promotion of ACE2-Ang（1-7）-MASR axis ，which leads to the delaying of the progression of chronic renal failure.

The role of licorice DXS knockout and overexpression in glycyrrhizic acid biosynthesis / 药学学报

1-Deoxy-D-xylulose-5-phosphate synthase (DXS) is a rate-limiting enzyme involved in the 2-C-methyl-D-erythritol 4-phosphate (MEP) pathway for terpenoid precursor biosynthesis. DXS plays an essential role in glycyrrhizic acid (GA) biosynthesis. Based on our previous transcriptome study, there was a negative correlation between DXS expression and GA content. Therefore, we explored the regulatory role of DXS in GA biosynthesis using both gene overexpression and gene knockout in a hairy root culture system. DXS was cloned from Glycyrrhiza glabra L. (GenBank Accession No. MN158121). A plant binary expression vector pCA-DXS was constructed by a gene fusion method. The sgRNA sequence was designed based on the first exon of DXS to construct the gene editing vector pHSE-DXS. Hairy roots overexpressing or knocking out DXS were generated through an Agrobacterium-mediated method with licorice hypocotyls as explants. Wild-type hairy roots and negative control hairy roots containing empty plasmids were also evaluated. UPLC was used to determine the GA content in each licorice hairy root line. Results showed that the content of GA in the hairy root group knocking out DXS was significantly higher than that in the wild-type and negative control groups, while in the hairy root group overexpressing DXS was significantly lower, suggesting that DXS plays a negative role in GA biosynthesis. This study provides a foundation for determining the function of DXS in terpenoid metabolism and for further establishment of a molecular regulatory network of GA biosynthesis.

Overexpressing of chalcone isomerase (CHI) gene enhances flavonoid accumulation in Glycyrrhiza uralensis hairy roots / 药学学报

Chalcone isomerase (CHI) is the second rate-limiting enzyme involved in the biosynthetic pathway of flavonoids in Glycyrrhiza uralensis. Based on our previous studies, we selected the specific CHI haplotype (GenBank Accession No. KY115232) to maximize flavonoid accumulation. We constructed a plant binary expression vector for overexpression of this CHI gene by the gene fusion method and transfected the plasmid into Agrobacterium tumefaciens ACCC10060 by electroporation. The recombinant A. tumefaciens ACCC10060 subsequently was used to infect cotyledons and hypocotyls of G. uralensis to obtain transgenic hairy roots. A qRT-PCR method was used to determine the copy number of CHI and a UPLC method was used to assay the content of four flavonoids in different hairy root lines. The qRT-PCR results showed that the copy number of CHI in hairy roots was 1 or 5. UPLC results showed that the content of total flavonoids, liquiritin, liquiritigenin, and isoliquiritigenin in transgenic hairy root samples was significantly higher than that in wild-type samples. This study demonstrates that overexpression of CHI significantly increases the content of flavonoids in hairy roots of G. uralensis. This work provides a theoretical basis for clarifying the function of CHI. Three transgenic hairy root lines of G. uralensis were isolated which can be used to increase the accumulation of licorice flavonoids in vitro.

Molecular Cloning and Sequence Analysis of GgUGE Gene in Glycyrrhiza glabra / 中国实验方剂学杂志

Objective::To clone the cDNA sequence of UDP-glucose 4-epimerase (UGE) in Glycyrrhiza glabra and analyze its sequence, so as to explore the potential relationship between the UGE gene and the molecular regulatory mechanisms of glycyrrhizic acid biosynthesis. Method::The cDNA sequence of UGE was cloned from the root of G. glabra by reverse transcription polymerase chain reaction (RT-PCR), then sequenced and analyzed by bioinformatics software. Results::A GgUGE cDNA sequence with the full length of 1 121 bp was obtained. The open reading fame (ORF) of GgUGE was 1 053 bp, encoding 350 amino acid residues. The GgUGE cDNA sequence was submitted to GenBank, and the accession No. was MK638908. Sequence analysis showed that GgUGE was an unstable hydrophilic protein, its average relative molecular weight was 39.02 kDa, and isoelectric point was 6.13. It contained no signal peptides or transmembrane domains. Its secondary structure mainly constituted of α-helix and had a conversed domain of UDP-glucose 4-epimerase superfamily. The homologoue analysis showed that the cDNA and amino acid sequences of GgUGE had the closest evolutionary relationship to Leguminosae and relatively distant evolutionary relationship to Salicaceae. Conclusion::In this study, GgUGE cDNA sequence is successfully cloned from G. glabra for the very first time, which will provide reference for studying the function of GgUGE and explaining the molecular regulatory mechanisms of glycyrrhizic acid biosynthesis in G. glabra.