The Expression and Correlation of miR-195, miR-125 and Calreticulin in Diffuse Large B-Cell Lymphoma / 中国实验血液学杂志

OBJECTIVE@#To analyze the expression and correlation of microRNA-195 (miR-195), miR-125 and calreticulin in diffuse large B-cell lymphoma (DLBCL).@*METHODS@#From April 2020 to April 2021, 80 DLBCL patients with complete data archived by the Pathology Department of Handan First Hospital and The Second Hospital of Hebei Medical University were selected as the study group, and 70 patients with reactive lymph node hyperplasia were selected as the control group. The expressions of miR-195 and miR-125 were detected by real-time fluorescence quantitative PCR, and the expression of calreticulin was detected by Western blot. Pearson correlation was used to analyze the correlation between miR-195, miR-125, calreticulin and DLBCL, and ROC curve was used to analyze the predictive value of miR-195, miR-125 and calreticulin for DLBCL.@*RESULTS@#Compared with the control group, the expression of miR-195 decreased but miR-125 and calreticulin increased in the study group (P<0.001). The expression levels of miR-195, miR-125 and calreticulin were not related to sex, age, primary site and B symptoms of patients with DLBCL, but related to immunophenotype, Ann Arbor stage, lactate dehydrogenase, IPI score, nodule involvement and Ki-67 index. The expression of miR-195 decreased and the expression of miR-125 and calreticulin increased in DLBCL paitents with non-germinal center source, Ann Arbor stage III-IV, lactate dehydrogenase > 245 U/L, IPI score 3-5, nodule involvement≥2 and Ki-67 index≥75% (P<0.05). Pearson correlation analysis showed that miR-195 and miR-125 were negatively correlated (r=-0.536, P=0.001), miR-195 and calreticulin were negatively correlated (r=-0.545, P=0.001), while miR-125 and calreticulin were positively correlated (r=0.523, P=0.001). ROC curve showed that compared with the single diagnosis of miR-195, miR-125 and calreticulin, the combination of the three items had higher predictive value for DLBCL (P<0.001).@*CONCLUSION@#The expression of miR-195 decreases and the expression of miR-125 and calreticulin increase in patients with DLBCL. Along with the increase of disease stage and IPI score, the decrease of miR-195 and the increase of miR-125 and calreticulin aggravate gradually. The three items may participate in the occurrence and progress of DLBCL.

Association of intrauterine benzo[a]pyrene exposure with benzo[a]pyrene diolepoxide (BPDE)-DNA adduct levels and pancreatic functional impairment in offspring rats / 预防医学

Objective@#To investigate the benzo[a]pyrene ( B[a]P ) diolepoxide ( BPDE )-DNA adduct levels in offspring rats with intrauterine exposure to B[a]P, and examine the effects of BPDE-DNA adduct levels on pancreatic functional impairment and glucose metabolism in offspring rats. @*Methods@#Forty pregnant rats were randomly divided into the blank control group, standard-dose group, low-dose group, medium-dose group and high-dose group (daily dose of 0, 2, 200, 800, 1 600 μg/kg B[a]P, respectively), of 8 animals in each group. Rats in the B[a]P treatment groups were administered by oral gavage with a mixture of B[a]P and corn oil at a dose of 0.2 mL/100 g body weight since day 1 of pregnancy until 21 days after delivery, while rats in the blank control group were given the same volume of coin oil by oral gavage. The BPDE-DNA adduct levels were measured and the pancreatic development was observed in the offspring rats 2 and 21 days and 12 weeks after birth, and the correlation between pancreas volume index and dose of exposure to B[a]P was examined using Spearman's rank correlation analysis. In addition, glucose metabolism was measured in offspring rats 12 months after birth using glucose tolerance test ( GTT ) and insulin tolerance test ( ITT ). @*Results@#There was no abnormal appearance, death, abortion or preterm birth in pregnant or offspring rats in the five groups, and no significant differences were seen in activity, diet, drinking water or mental status in rats. The greatest level of BPDE-DNA adducts was measured in offspring rats 2 days after birth, with median levels ( interquartile range ) of 1 089.60 ( 586.10 ) to 1 405.49 ( 346.47 ) pg/mL, and no BPDE-DNA adducts were found in offspring rats 12 weeks after birth. The pancreas volume index correlated negatively with the dose of exposure to B[a]P in offspring rats 2 ( rs=-0.620, P=0.001 ) and 21 days after birth ( rs=-0.801, P=0.001 ). Hypoplasia of pancreas with loose tissues was seen in offspring rats 2 days after birth, while well pancreatic development was found in offspring rats 12 weeks after birth, with tight exocrine portion. GTT showed an increase in glucose levels in offspring rats in all five groups following abdominal injection of glucose and declined 30 min post-injection ( F=365.578, P<0.001 ), and ITT showed a tendency towards a decline in glucose levels in offspring rats in all five groups ( F=461.215, P<0.001 ).@*Conclusions@#The levels of BPDE-DNA adducts in offspring rats increase with the dose of intrauterine B[a]P exposure, and insulin resistance and impaired glucose tolerance occur 12 months post-exposure to B[a]P. Intrauterine B[a]P exposure affects pancreatic development in offspring rats and causes abnormal glucose metabolism in adult offspring rats.

Establishment of multiple evidence-integrated evaluation and prediction method for "toxic" Chinese medicines / 中国中药杂志

Traditional Chinese medicine(TCM) carries the experience and theoretical knowledge of the ancients, and the use of "toxic" Chinese medicines is a major feature and advantage of TCM. "Toxic" Chinese medicines have unique clinical value and certain medication risk under the guidance of TCM theories such as compatibility for detoxification and treatment based on syndrome differentiation. In recent years, the safety events of Chinese medicines have occurred frequently, which has made the safety of Chinese medicine a public concern in China and abroad. However, limited by conventional cognitive laws and technical methods, basic research on toxicity of Chinese medicines fails to be combined with the clinical application. As a result, it is difficult to identify the clinical characteristics of, predict toxic and side effects of, or form a universal precise medication regimen for "toxic" Chinese medicines, which restricts the clinical application of them. In view of the problem that the toxicity of "toxic" Chinese medicines is difficult to be predicted and restricts the clinical application, the evidence-based research concept will provide new ideas for safe applcation of them in clinical practice. The integrated development of multiple disciplines and techniques in the field of big data and artificial intelligence will also promote the renewal and development of the research models for "toxic" Chinese medicines. Our team tried to propose the academic concept of evidence-based Chinese medicine toxicology and establish the data-intelligence research mode for "toxic" Chinese medicines and the intelligent risk prediction method for medicinal combination in the early stage, which provided methodological supports for solving the above problem. Thus, on the basis of summarizing the research status and problems of the clinical medication regimen of "toxic" Chinese medicines, our team took the evidence-based toxicology of TCM as the core concept, and tried to construct the multiple-evidence integrated evaluation and prediction method for "toxic" Chinese medicine, so as to guide the establishment of the non-toxic medication regimen of "toxic" Chinese medicines. Specifically, through the analysis of multivariate data obtained from the basic research, the evidence-based toxicology database of Chinese medicines and the individualized "toxicity-effect" intelligent prediction platform were built based on the disease-syndrome virtual patients, so as to identify the clinical characteristics and risks of "toxic" Chinese medicines and develop individualized medication regime. This study is expected to provide a methodological reference for the establishment of medication regimen and risk prevention strategy for "toxic" Chinese medicines. The method established in this study will bridge clinical research and basic research, enhance the transformation of the scientific connotation of attenuated compatibility, promote the development of evidence-based Chinese medicine toxicology, and ensure the clinical safety of "toxic" Chinese medicines.

Risk factors for anastomotic leakage after laparoscopic lower anterior resection of rectal cancer and application value of risk assessment scoring model: a multicenter retrospective study / 中华消化外科杂志

Objective:To investigate the risk factors for anastomotic leakage after laparo-scopic lower anterior resection (LAR) of rectal cancer, and the application value of its risk assess-ment scoring model.Methods:The retrospective case-control study was conducted. The clinico-pathological data of 539 patients who underwent laparoscopic LAR of rectal cancer in 13 medical centers, including 248 cases in Renji Hospital of Shanghai Jiaotong University School of Medicine, 35 cases in Ningbo First Hospital, 35 cases in Changzhou Second People's Hospital, 32 cases in the First People's Hospital of Nantong, 32 cases in Linyi People's Hospital, 31 cases in Changzhou Wujin People's Hospital, 28 cases in Jiading District Hospital of Traditional Chinese Medicine, 27 cases in the First Hospital of Taizhou, 26 cases in Shanghai Pudong Gongli Hospital, 21 cases in the People's Hospital of Rugao, 11 cases in Central Hospital of Fengxian District, 7 cases in Ningbo Hangzhou Bay Hospital and 6 cases in Jiangsu jianhu People's Hospital, from January 2016 to November 2020 were collected. There were 157 males and 382 females, aged (62.7±0.5)years. Observation indicators: (1) follow-up; (2) risk factors for anastomotic leakage after laparoscopic LAR; (3) establishment of risk assessment scoring model for anastomotic leakage after laparoscopic LAR. Follow-up was conducted by outpatient examination or telephone interview. Patients were followed up at 1 week after discharge or 1 month after the operation to detect the anastomotic leakage. Measurement data with normal distribution were represented as Mean± SD, and measurement data with skewed distribution were represented as M(range). Count data were represented as absolute numbers or percentages, and comparison between groups was analyzed using the chi-square test. Univariate analysis was conducted using the chi-square test and multivariate analysis was conducted usong the Logistic regression model. The area under curve of receiver operating characteristic curve was used to estimate the efficiency of detecton methods. The maximum value of the Youden index was defined as the best cut-off value. Results:(1) Follow-up: 539 patients were followed up at postoperative 1 week and 1 month. During the follow-up, 79 patient had anastomotic leakage, with an incidence of 14.66%(79/539). Of the 79 patients, 39 cases were cured after conservative treatment, 40 cases were cured after reoperation (ileostomy or colostomy). (2) Risk factors for anastomotic leakage after laparoscopic LAR. Results of univariate analysis showed that sex, age, body mass index, smoking and/or drinking, tumor diameter, diabetes mellitus, hemoglobin, albumin, grade of American Society of Anesthesio-logists (ASA), neoadjuvant chemoradiotherapy, distance from anastomotic level to dentate line, the number of pelvic stapler, reinforced anastomosis, volume of intraoperative blood loss, placement of decompression tube, preservation of left colic artery, operation time and professional doctors were related factors for anastomotic leakage after laparoscopic LAR ( χ2=14.060, 4.387, 5.039, 4.094, 17.488, 33.485, 25.066, 28.959, 34.973, 34.207, 22.076, 13.208, 16.440, 17.708, 17.260, 4.573, 5.919, 5.389, P<0.05). Results of multivariate analysis showed that male, tumor diameter ≥3.5 cm, diabetes mellitus, hemoglobin <90 g/L, albumin <30 g/L, grade of ASA ≥Ⅲ, neoadjuvant chemoradiotherapy, distance from anastomotic level to dentate line <1 cm, the number of pelvic stapler ≥3, non-reinforced anastomosis, volume of intraoperative blood loss ≥100 mL and no placement of decom-pression tube were independent risk factors for anastomotic leakage after laparoscopic LAR ( odds ratio=2.864,3.043,12.556,7.178,8.425,12.895,8.987,4.002,3.084,4.393,3.266,3.224,95% confidence interval as 1.279?6.411, 1.404?6.594, 4.469?35.274, 2.648?19.459, 2.471?28.733, 4.027?41.289, 3.702?21.777, 1.746?9.171, 1.365?6.966, 1.914?10.083, 1.434?7.441, 1.321?7.867, P<0.05). (3) Establishment of risk assessment scoring model for anastomotic leakage after laparoscopic LAR. based on the results of univariate analysis, clinicopathological factors with χ2>20, χ2>10 and ≤20 or χ2≤10 were defined as scoring of 3, 2, 1, respectively. The cumulative clinicopatho-logical factors scoring ≥6 was defined as an effective evaluating indicator for postoperative anastomotic leakage. The risk assessment scoring model (6-321) for anastomotic leakage after laparoscopic LAR was established. The cumulative value ≥6 indicated high incidence of anastomotic leakage, and the cumulative value <6 indicated low incidence of anastomotic leakage. Conclusions:Male, tumor diameter ≥3.5 cm, diabetes mellitus, hemoglobin <90 g/L, albumin <30 g/L, grade of ASA ≥Ⅲ, neo-adjuvant chemoradiotherapy, distance from anastomotic level to dentate line <1 cm, the number of pelvic stapler ≥3, non-reinforced anastomosis, volume of intraoperative blood loss ≥100 mL and no placement of decompression tube are independent risk factors for anastomotic leakage after laparoscopic LAR. The risk assessment scoring model (6-321) is established according to the above results.The cumulative value ≥6 indicates high incidence of anastomotic leakage and the cumulative value <6 indicates low incidence of anastomotic leakage.

Relationship between Polymorphism of miR-155 and Its Target Gene MyD88 and Clinicopathological Features of Diffuse Large B-cell Lymphoma / 中国实验血液学杂志

OBJECTIVE@#To investigate the relationship between the polymorphism of miR-155 and its target gene MyD88 and clinicopathological features of diffuse large B-cell lymphoma (DLBCL).@*METHODS@#135 cases of DLBCL patients in our hospital from March 2015 to August 2017 were selected, and 90 cases of reactive hyperplasia of lymph nodes were selected as the control group. The relative expression of miR-155 and MyD88 gene polymorphism were detected in the two groups, and the relationship between miR-155 and MyD88 gene polymorphism and clinicopathological characteristics of DLBCL was analyzed.@*RESULTS@#The relative expression of miR-155 in DLBCL patients was significantly higher than that in the control group (P<0.05). The mutation rate of MyD88 L265P in DLBCL group was significantly higher than that in control group (P<0.05). The relative expression of miR-155 in patients with MyD88 L265P mutation was significantly higher than that in patients with wild-type DLBCL (P<0.05). The relative expression of miR-155 and the polymorphism of MyD88 L265P were associated with lesion location, stage, BCL-2 protein expression and MyD88 protein expression in DLBCL patients (t=7.461、8.804、6.487、10.812； χ@*CONCLUSION@#The abnormal expression of miR-155 and the mutation rate of MyD88 gene in DLBCL patients are increased, and the expression of miR-155 and the mutation of MyD88 gene affect the disease progression and prognosis of patients, which may be potential biological indicators for the diagnosis, treatment and prognosis of DLBCL.

Effects of Long Non-coding RNA-TUC338 on the Migration and Proliferation of Lymphoma Cells via PI3K/AKT Signaling Pathway / 中国实验血液学杂志

OBJECTIVE@#To investigate the effect of long non-coding RNA-TUC338 on the proliferation and migration of lymphoma cells.@*METHODS@#The expression of TUC338 in different lymphoma cells was detected by fluorescence quantitative PCR, cell proliferation by sulforhodamine B (SRB) assay, migration of lymphoma cells by transwell assay, and protein expression in PI3K/AKT signaling pathway by Western blot.@*RESULTS@#The expression levels of TUC338 in lymphoma cells Daudi, U937, BC-3, and Raji significantly increased in comparison with human normal T lymphocytes H9 (t=13.277, 10.103, 16.200, and 26.687, P=0.002, 0.005, 0.001, and 0.000). Compared with NC-siRNA group, the number of cells crossing the chamber of TUC338-siRNA group was significantly reduced (t=30.508, P=0.000), the protein expression levels of p-PI3K and p-AKT significantly decreased (t=16.872 and 18.371, P=0.000 and 0.000), and OD@*CONCLUSION@#The expression of TUC338 significantly increases in lymphoma cells, and silence of TUC338 effectively inhibits the activation of PI3K/AKT signaling pathway, thereby inhibiting the proliferation and migration of lymphoma cells, which has a potential application value in diagnosis and treatment of lymphoma.

Determination of UV-327 and UV-328 in mouse plasma by high performance liquid chromatography / 北京大学学报(医学版)

OBJECTIVE@#To establish a high performance liquid chromatography (HPLC) method for the determination of ultraviolet (UV) absorbers UV-327 and UV-328 in mouse plasma.@*METHODS@#N-hexane-acetone (volume ratio 1 ∶1) was added to a mouse plasma sample as the extraction solvent for vortex extraction, and the supernatant was dried at 50 ℃ with nitrogen. Thereafter the residue was redissolved with methanol, centrifuged and filtered. The separation was performed on a Waters SymmetryC18 column (250 mm×4.6 mm, 5 μm), and the concentrations of UV-327 and UV-328 in the mouse plasma were determined by HPLC with an UV detector. The elution was isocratic at a flow rate of 1.0 mL/min with a mobile phase composed of 100% methanol, and the UV detection wavelength was 340 nm. The retention time was used for qualitative analysis, and the internal standard method was used for quantitative analysis using UV-320 as the internal standard.@*RESULTS@#The calibration curves of UV-327 and UV-328 were linear with correlation coefficients of 0.999 7 over the concentration range of 0.05 to 10.0 mg/L. The limit of detection was 0.01 mg/L, and the limit of quantitation was 0.03 mg/L. The average recoveries at low, medium and high three concentrations (0.50, 1.00, 2.00 mg/L) in the mouse plasma were 91.7%-101.0% for UV-327, and 97.5%-103.9% for UV-328. The intra-day precisions (n=6) of UV-327 were 2.9%-6.6%, and 2.7%-7.4% for UV-328. The inter-day precisions (n=3) of UV-327 were 6.0%-9.3%, and 6.6%-8.6% for UV-328. The extraction recoveries of UV-327 were 98.8%-103.8%, and 99.8%-100.9% for UV-328. The measured relative deviations of UV-327 in the mouse plasma samples placed at room temperature for 6 hours and -40 ℃ for 15 days were 0.9%-3.5% and 7.4%-15.0%, and the measured relative deviations of UV-328 were 2.0%-4.3% and 2.1%-13.8%, respectively. The mouse plasma samples could be stored at room temperature for 6 hours at least and -40 ℃ for 15 days at three spiked concentration levels.@*CONCLUSION@#The method was simple and fast with high accuracy, precision and sensitivity, and could be applied to the determination of UV-327 and UV-328 in mouse plasma.

Advances in Evaluation Methods and Models on Angiogenesis / 中国药学杂志

Angiogenesis, the process of shaping new vessels from existing or posteriorvessels. It occurs mainly after birth and is the basis for the formation of many vascular structures and participates ina variety of physiological and pathological processes.Due to the particularity of the research objects and the diversity of the species, the experimental results obtained from one model may not be confirmed on the other model. At present,the comprehensive evaluation of angiogenesis requires mutual verification of different model results. Herein, this review mainly summarizes the literatures on the evaluation of angiogenesis activity within recent ten years. From three parallel dimensions of cell, tissue, and animal,the up-to-date evaluation methods and angiogenesis models with wide recognitions are reviewed, in order to provide the reference for related drug research and development.

Systematic Review of Jinkui Shenqi Pills on Hypertension / 中国中医药信息杂志

Objective To evaluate the efficacy and safety of Jinkui Shenqi Pills on hypertension through Meta-analysis and systematic review. Methods Articles about intervention of modified Jinkui Shenqi Pills in hypertensive clinical randomized controlled trials were retrieved from CNKI, Wanfang database, VIP, Cochrane Library, PubMed, and Embase. They were all searched from inception to June 2016. Cochrane system evaluation manual bias risk assessment method was used to evaluate the quality of the articles, and RevMan5.3 software was used for Meta-analysis. Results Nine articles involving 776 cases were included in the study. Meta-analysis showed that the total effective rate of modified Jinkui Shenqi Pills was better than that of conventional Western medicine treatment [OR=2.67, 95%CI (1.73, 4.11), P＜0.000 01], and combination therapy was superior to conventional Western medicine alone [OR=3.27, 95%CI (1.99, 5.39), P＜0.000 01]. However, the efficacy of modified Jinkui Shenqi Pills alone was equivalent to the conventional Western medicine alone [OR=1.16, 95%CI (0.43, 3.09), P=0.77). Conclusion Modified Jinkui Shenqi Pills can improve the clinical symptoms of the patients with hypertension, and combination treatment can improve the antihypertensive effect. However, the quality of the included research literature is too low, yet more large-scale, multi-center, high-quality clinical randomized controlled trials are needed to further verify.

Evaluation of cytotoxicity of stearoyl gemcitabine-loaded PLGA nanoparticles to Lewis lung cancer cells / 中国免疫学杂志

Objective:To investigate the anti-proliferation effect of 4-(N)-stearoyl gemcitabine-loaded poly(lactic-co-glycolic) acid nanoparticles(GemC18-PLGA-NPs) on Lewis lung cancer cells(LLC) in vitro.Methods: Lewis cells were incubated with GemC18-PLGA-NPs,free GemC18,gemcitabine HCl(GemHCl) or GemC18-free blank nanoparticles(PLGA-NPs) respectively and cell viability was determined using an MTT assay after 24,48 or 72 h of incubation.The apoptosis rate after 48 and 72 h of incubation were measured by flow cytometry.Results:GemC18-PLGA-NPs,GemC18,and GemHCl all significantly inhibited the growth of LLC cells, and the survival rate of GemHCl group was lowest,GemC18-PLGA-NPs group had the highest survival rate.The cell survival rate of GemC18-PLGA-NPs after 72 h was significantly higher than that of GemHCl (P<0.05) at the concentration of 1 μmol/L,indicating that it had a significant drug release effect.PLGA-NPs group produced trifle inhibition on the Lewis cells without correlation to time or concentration.Conclusion:GemC18-PLGA-NPs have significant anti-proliferation effect on mouse Lewis lung cancer cells in vitro.

Application of perineural space expansion by normal saline in axillary brachial plexus block / 上海交通大学学报（医学版）

Objective· To compare the clinical effect of ultrasound-guided perineural space expansion by normal saline combined with perivascular technique (PSE-PV) with traditional perivascular technique in axillary brachial plexus block (PV).Methods· A randomized,controlled,and double blind study was performed.Sixty patients arranged for emergency hand surgery were allocated to 2 groups,i.e.PSE-PV group and PV group,with 30 cases in each group receiving ultrasound-guided axillary brachial plexus block.For PSE-PV group,injection was carried out at the 12 o'clock position using 5 mL of normal saline firstly to expand the perineural space followed by 15 mL of 0.5％ ropivacaine at the same position.The patients of PV group got their injections of 20 mL of 0.5％ ropivacaine at the direction of 12 o'clock of axillary artery.The sensory and motor blockade assessment 30 min after drug injection,performance time,success rate,and adverse events were recorded.Results· The sensory and motor blockade scores were 1.7±0.3 and 1.6±0.3 in PSE-PV group,and 1.8±0.4 and 1.6±0.3,respectively in PV group without significant differences (P＞0.05).The performance time of PSE-PV group was (4.0± 1.3) min,a little longer than that of PV group,but the difference was not statistically significant (P＞0.05).The success rates of nerve block reached 100％ in both groups.Only l case in PV group reacted to the local anesthetics.Conclusion · Ultrasound-guided perineural space expansion by normal saline reduces dosage of local anesthetics in axillary brachial plexus block,which has similar anesthetic effect,performance time and adverse effect with traditional perivascular method.

Advances on anti-neoplastic STAT3 inhibitors / 药学学报

Signal transducer and activator of transcription 3 (STAT3) is a kind of signal transduction protein involved in cell proliferation, differentiation, apoptosis and other important physiological processes in response to a large number of cytokines and growth factors in cells. It has been shown that constitutive activation of STAT3 is closely associated with oncogenesis and tumorigenesis. Inhibition of aberrant STAT3 signaling has been one of promising strategies for the development of anti-neoplastic therapeutics. The review summarizes the latest progress of STAT3 inhibitors in recent years from the perspective of targeting N-terminal domain, DNA binding domain, SH2 domain and C-terminal transactivation domain of STAT3.

Advances in the pharmacological function of FBPase and development of FBPase inhibitors / 药学学报

Fructose 1, 6-bisphosptase (FBPase), a second rate-limiting enzyme in gluconeogenesis, has an important role in the control of gluconeogenesis, which involves in energy metabolism and glucose homeostasis. Inhibitors of FBPase exhibit an anti-diabetic activity. Some of FBPase inhibitors have entered the stage of clinical trials, which indicates that FBPase is a promising therapeutic target for the discovery and development of hypoglycemic drugs. In addition, recent studies have shown that FBPase can be used to treat other diseases such as the initiation and development of tumors in several cancer types. Here, we provide a review of the biological characteristics of FBPase and contributions of FBPase on gluconeogenesis and insulin secretion, the research and development of FBPase inhibitors and the regulatory role of FBPase in other diseases.

Determination of 8-methoxypsoralen in mouse plasma by high performance liquid chromatography and its application to pharmacokinetic study / 北京大学学报(医学版)

OBJECTIVE@#To establish a high performance liquid chromatography (HPLC) method for the determination of 8-methoxypsoralen (8-MOP) in mouse plasma and apply it to a pharmacokinetic study of 8-MOP.@*METHODS@#8-MOP was separated on a Waters Symmetry18 column (250 mm × 4.6 mm, 5 μm) and determined by HPLC using isocratic elution, and 5-methoxypsoralen was used as internal standard. The mobile phase consisted of methanol-water (55:45, V/V) at a flow rate of 1.0 mL/min. The excitation and emission wavelength of fluorescence detector were set at 334 nm and 484 nm respectively, and the internal standard method was used for quantitative analysis. In the study, 60 healthy ICR male mice were randomly divided into twelve groups. The mice in control group were administered intragastrically with 1% Tween 80, and the mice in the other eleven groups were administered intragastrically with 8-MOP (40 mg/kg). Plasma concentrations of 8-MOP in the mice at different time points after treatment were determined by HPLC. Pharmacokinetic parameters were calculated by DAS 2.0 software.@*RESULTS@#The calibration curve of 8-MOP was linear with a correlation coefficient of 0.999 3 over the concentration range of 0.05 to 10 mg/L, and the limit of detection was 0.015 mg/L. The average recoveries of 8? MOP at three different concentrations (0.10, 0.50, 2.5 mg/L) were from 92.5% to 100.6%. The intra-day precision of 8-MOP was from 3.3% to 8.2%, while the inter-day precision was from 3.4% to 6.7% at three spiked concentration levels. The extraction recoveries of 8-MOP were from 90.9% to 92.0%, and the plasma samples could be stored at -80°C for 15 days at least at three spiked concentration levels. 8-MOP could be detected in mouse plasma 5 min after intragastrical administration to the mice (1.4 mg/L). The concentration of 8-MOP in the mouse plasma reached a maximum 2 h after administration, and 8-MOP could still be detected 24 h after administration (1.1 mg/L). t1/2 was (39.21±3.65) h, Cmax was (2.31±0.02) mg/L, tmax was (2.00±0.00) h, and AUC0-t was (33.34±1.19) (h×mg)/L.@*CONCLUSION@#The proposed method is accurate and simple,suitable for pharmacokinetics of 8-MOP in mice.

Expression and Clinical Significance of LncRNA KCNQ1OT1 in Patients with Acute Myeloid Leukemia / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the expression of LncRNA KCNQ1OT1 in patients with acute myeloid leukemia (AML) and to analyze the relation of LncRNA KCNQ1OT1 expression levels with clinicopathological features.</p><p><b>METHODS</b>A total of 68 patients with AML were enrolled in the study, 48 out of them were suffered from acute myeloid leukemia (AML) and 20 reached to complete remission (CR), 30 age-matched patients with iron-deficient anemia were included in control group, the peripheral blood samples of all the patients were collected, and the real-time fluorescent quantitative PCR (qRT-PCR) was used to detect the expression of LncRNA KCNQ1OT1, meanwhile, the correlation of its expression with clinicopathological characteristics and prognosis was analyzed.</p><p><b>RESULTS</b>The expression of LncRNA KCNQ1OT1 in AML patients was significantly higher than that in the patient with complete remission and iron-deficient anemia (F=14.67, P<0.01). The expression of LncRNA KCNQ1OT1 was not significantly different between 20 cases of AML-CR and 30 cases of iron-deficient anemia (P>0.05). The expression of LncRNA KCNQ1OT1 was associated with NCCN risk grade and survival status in patients with AML. The median overall survival time was significantly shorter in patients with high expression of LncRNA KCNQ1OT1 than that in patients with low expression(P<0.05).</p><p><b>CONCLUSION</b>LncRNA KCNQ1OT1 may be involved in the regulation of AML. Expression of LncRNA KCNQ1OT1 and NCCN risk score can be used as biomarkers of prognosis in the patients with AML and may be a potential prognostic marker and therapeutic target for AML patients.</p>

Application of indocyanine green in sentinel lymph node biopsy of breast cancer and analysis of dose-response / 上海交通大学学报（医学版）

Objective·To study the imaging of subcutaneous lymph-vessels and lymph-nodes in sentinel lymph node biopsy (SLNB) with different concentrations of indocyanine green (ICG), and to find the best concentration to improve the success rate of SLNB. Methods·A total of 339 primary breast cancer patients with clinical negative axillary lymph node were enrolled in the study. Double tracer method [ICG and methylene blue (10 mg/mL)] was used for sentinel lymph node biopsy. 1.25 mg/mL ICG was diluted to difference concentration. The injection volume was 1 mL for ICG and 1 mL for methylene blue. The imaging of subcutaneous lymph-vessels and lymph-nodes and the success rate of SLNB were recorded after injection of different concentrations of ICG. Results·The concentration of ICG had significant effect on lymph-vessels and lymph-nodes imaging rate and SLNB success rate (P=0.001). In the group that breast tumors not located in the external upper quadrant (needle biopsy or excisional biopsy) and tumor located in the external upper quadrant without excisional biopsy (needle biopsy), concentration of ICG reached 80％ subcutaneous lymph-vessels imaging rate is 0.723 mg/mL (95％ CI 0.595-0.915 mg/mL). In this group, in this concentration range, the imaging rate of subcutaneous lymph node was 28％-53％, and the success rate of SLNB was 88％-92％. In the group that breast tumors located in the external upper quadrant and had been resected, the imaging rate of subcutaneous lymph-vessels and lymph-nodes was 50％-69％ and 20％-42％, respectively, and the success rate of SLNB was 76％-81％. The difference of subcutaneous lymph-vessels imaging rate between two groups was statistically significant (P=0.014), but the difference of lymph-nodes imaging rate and SLNB success rate had no significance (P=0.628, P=0.232). Conclusion·Using double tracer (ICG and methylene blue) in SLNB is simple and feasible. The best concentration of ICG is 0.723 mg/mL, which can improve the imaging rate of lymph nodes and lymph nodes.

Pro-apoptotic effect of a tumor suppressor gene WWOX on ovarian cancer stem cells and its mechanism / 中国组织工程研究

BACKGROUND:WWOX,a tumor suppressor gene,can affect the growth of ovarian cancer stem cells;however,there is no report on whether its mechanism of action is related to Hedgehog signaling pathway.OBJECTIVE:To investigate the effect and mechanism of overexpression of WWOX on the apoptosis of ovarian cancer stem cells.METHODS:pcDNA3.1-WWOX (pcDNA3.1-WWOX group) and pcDNA4.0-WWOX (pcDNA4.0-WWOX group) were transferred into ovarian cancer stem cells,respectively;and meanwhile,pcDNA3.1 (pcDNA3.1 group) and pcDNA4.0 (pcDNA4.0 group) were transferred into the cells.A non-transfection group (only with Lipofectamine2000) was set up.After cultured 48 hours,the levels of WWOX in the pcDNA3.1-WWOX group and pcDNA4.0-WWOX group were detected using western blot assay,and the cell proliferation and apoptosis were detected using MTT assay and flow cytometry,respectively.Western blot assay was also used to detect the levels of Hedgehog signaling pathway associated proteins,SHH,PTCH1,Gli-1,SMO and apoptosis-related protein Cleaved Caspase-3 in the cells.Cyclopamine,Hedgehog signaling pathway inhibitor,was used in ovarian cancer stem cells without transfection (cyclopamine group) and after the transfection of WWOX overexpression vector (WWOX+cyclopamine group) followed by 48 hours of culture,and then MTT,flow cytometry and western blot detections were performed.RESULTS AND CONCLUSION:(1) The expression level of WWOX in the pcDNA4.0-WWOX group was significantly higher than that in the pcDNA3.1-WWOX group (t=27.84,P=0.00).The ovarian cancer stem cells which were transfected with pcDNA4.0-WWOX were used to overexpress WWOX in the late experiment.(2) Overexpression of WWOX could inhibit the proliferation of ovarian cancer stem cells and promote the apoptosis of ovarian cancer stem cells.(3) Overexpression of WWOX could inhibit the expression of Gii-1,PTCH1,SMO and SHH in ovarian cancer stem cells,and promote the expression of Cleaved Caspase-3.(4) Cyclopamine could inhibit the expression of SHH,PTCH 1,Gli-1,SMO,and promote the expression of Cleaved Caspase-3.Cyclopamine had obvious inhibitory effect on Hedgehog signaling pathway.(5) Cyclopamine could enhance the apoptosis induced by overexpression of WWOX in ovarian cancer stem cells,and enhance the inhibition of proliferation of ovarian cancer stem cells induced by overexpression of WWOX.To conclude,WWOX effects on proliferation and apoptosis of ovarian cancer stem cells may be related to the inhibition of Hedgehog signaling pathway.

nhibitory effect and mechanism of deoxyschizandrin on NLRP3 inflammasome / 药学学报

This study was conducted to investigate the inhibitory effect and the molecular mechanism of deoxyschizandrin on the activity of NLRP3 (NOD-like receptor family, pyrin domain containing 3) inflammasome. Bone marrow-derived macrophages were used to study the effects of deoxyschizandrin on inflammasome activation using inflammasome inducers (ATP and nigericin). Cytotoxic effect was evaluated with CCK-8. The expression of IL-1β, caspase-1 in the supernatant and the expression of pro-caspase-1, pro-IL-1 β, ASC, NLRP3 in cell was detected by Western blot for the inhibitory effect of deoxyschizandrin (25, 50, 100 and 200 μmol·L-1) on the activity of NLRP3 inflammasome. Immunofluorescence was applied to investigate NF-κB (p65) transportation to the nucleus. The results of CCK-8 showed that the optimum concentration of deoxyschizandrin was 6.25-400 μmol·L-1. Deoxyschizandrin (25, 50, 100, and 200 μmol·L-1) could inhibit the activation of NLRP3 inflammasome caused by nigericin and ATP, and inhibit the secretion of IL-1 β, which was associated with inhibiting the cleavage of pro-caspase-1. The results of immunofluorescence and Western blot also suggest that the inhibitory activity of deoxyschizandrin on NLRP3 inflammasome was not dependent on NF-κB pathway and protein expression of NLRP3, ASC, pro-caspase-1 and pro-IL-1 β mediated by NF-κB. Our results confirmed that deoxyschizandrin could suppress the cleavage of pro-caspase-1 and inhibit the activity of NLRP3 inflammasome at 25-200 μmol·L-1 to reduce the inflammation response.

The three dimensional organoids-based high content imaging model for hepatotoxicity assessment / 药学学报

The drug hepatotoxicity assessment method in vitro was established by 3D organoid model of HepaRG cell line in combination with high content imaging analysis. HepaRG cells were differentiated into hepatocyte-like morphology and bile canaliculus-like structures by treatment with hydrocortisone and dimethyl sulfoxide(DMSO), inducing the expressions of drug-metabolizing enzymes, transporters, nuclear receptors and hepatocyte-specific protein albumin(ALB)genes, finally forming the stable organoids with closely resembling liver function in vitro. Through the high content imaging analysis and the specific, multi-targets fluorescent dye, the number of live/dead cells, mitochondrial membrane potential(MMP), intracellular reactive oxygen species(ROS)were analyzed for the drug hepatotoxicity evaluation. The results showed that the organoids evaluation model of HepaRG cells in vitro could be used to assess accurately the difference between hepatotoxicity positive control drugs of amiodarone(AMD), cyclosporin(CSP)and the negative control drug of aspirin(ASP): AMD and CSP concentration-dependently decreased the number of total and live organoid cells. The number of dead organoid cells was increased sharply when the concentration of AMD was more than 50 μmol·L-1, while no significant changes was observed for ASP. AMD and CSP concentration-dependently caused the MMP declined and the ROS increased, with AMD showing a greater degree than CSP and ASP presenting no markedly effect. In conclusion, the organoid evaluation method of HepaRG cells in combination with high content imaging analysis can be used for the drug hepatotoxicity assessment in vitro. It displays the advantages of multi-target, high throughput, intuitive results as well as quantitatively.

Study on the mechanism of PPAR-γ dependent immunological idiosyncrasy liver injury induced by Polygonum multiflorum / 药学学报

To investigate the effects of peroxisome proliferator-activated receptor gamma(PPAR-γ)on the liver injury of Polygonum multiflorum, we established a model of immunological idiosyncrasy liver injury induced by lipopolysaccharide. The 70 Sprague-Dawley(SD)rats were randomly divided into control group, LPS group(2.8 mg·kg-1), PM group(crude drug, 2.16 g·kg-1), PPAR-γ agonist group(pioglitazone, 0.5 mg·kg-1), PM+LPS group(crude drug 2.16 g·kg-1, 2.8 mg·kg-1), PPAR-γ agonist+LPS group(0.5 mg·kg-1, 2.8 mg·kg-1)and PM+LPS+PPAR-γ agonist group(crude drug, 2.16 g·kg-1, 2.8 mg·kg-1, 0.5 mg·kg-1). The rats were orally given PM, once a day for consecutive 2 days. The control rats were given the same amount of distilled water. Liver injury was induced by intravenous injection of LPS. Sodium pentobarbital was injected intraperitoneally for anesthesia, and liver samples were collected together with blood. The plasma levels of alanine transaminase(ALT), aspartate aminotransferase(AST), tumor necrosis factor-α(TNF-α), interleukin-1β(IL-1β), interleukin-6(IL-6)and interferon-γ(IFN-γ)were measured. Pathological changes and hepatocellular apoptosis were examined by liver biopsy, and immunohistochemical observation of liver tissue expression of PPAR-γ and NF-κB p65. A negative correlation was observed between the expression of PPAR-γ in hepatic tissue and liver injury of Polygonum multiflorum. PPAR-γ agonist significantly reduced the PM-induced idiosyncratic liver injury in rats according to serum ALT and AST(P < 0.05), reduced liver pathological injury and hepatocyte apoptosis, decreased serum TNF-α and other inflammatory cytokines(P < 0.05), liver tissue PPAR-γ expression, and inhibited expression of NF-κB p65(P < 0.05). The results suggest that the occurrence of immunological idiosyncrasy liver injury of PM is related to inhibition of the PPAR-γ pathway and elevation of inflammatory factors. PPAR-γ agonist can reverse the idiosyncratic liver injury induced by PM, and provide a reference for elucidating mechanism of idiosyncratic liver injury induced by Polygonum multiflorum.