[CD19 antigen loss after treatment of Bispecific T-cell Engager and effective response to salvage bispecific CAR-T therapy in B cell acute lymphoblastic leukemia: a case report and literature review].

Objective: To analyze the influence of CD19 isoforms to the efficacy of CD19/CD3 Bispecific T-cell Engager (BiTE) antibody, and explore the resistance mechanism of BiTE immunotherapy. Methods: Semi-quantitative RT-PCR (qRT-PCR) was used to detect the expression of CD19 mRNA isoforms before and after BiTE treatment in a patient with CD19(+) B cell acute lymphoblastic leukemia (ALL) . CD19 isoforms were analyzed by Sanger sequencing. Flow cytometry and transcriptome sequencing were performed to analyze the expression of cell lineage specific molecules before and after BiTE treatment. Results: The expression of CD19 isoform with exon 2 deletion was identified at diagnosis. After relapsed and treatment of BiTE antibody, the patient did not achieve remission and CD19 antigen on leukemic cells turned negative detected by flow cytometry after BiTE treatment. However the expression ratio of CD19 isoform with exon 2 deletion was not increased. Flow cytometry phenotype and transcriptome sequencing confirmed that no linage switching developed, which suggested the expression of CD19 isoform caused by exon alternative splicing and lineage switching was not related to CD19 epitope loss in this patient. This patient achieved complete remission by sequential administration of self-developed CD22 CAR-T and CD19 CAR-T after disease progression. Conclusion: Targeting or combining an alternative antigen specific CAR-T may be a promising treatment option after losing CD19 expression in relapsed ALL.

[Retrospectively analysis of characteristics and risk factors of immune treatment-related hepatic adverse events in malignant tumor].

Objective: To explore the clinical features and risk factors of hepatic injury due to immune checkpoint inhibitors (CPI) therapy in malignant tumor. Methods: Data of 112 patients (64 men and 48 women) who received CPI between January 2016 and March 2019 in Chinese Academy of Medical Sciences and Peking Union Medical College Shenzhen Hospital, and Huazhong University of Science and Techology Union Shenzhen Hospital were retrospectively collected. The median age of these patients was 60 years. Results: Hepatic adverse events were observed in 30 patients out of 112 patients (26.8%). Among them, the incidence of grade 3-5 hepatic adverse events were 7.14% (8/112). The median time of hepatic adverse event occurrence was 3 weeks (2-30) after undergoing therapy. The results of univariate and multivariate analyses showed that liver cancer was attributed to the CPI induced hepatitis (P<0.05). Patients with severe hepatic injury got almost complete resolution after receiving methlprednisolone for 4 to 6 weeks. Conclusion: Live cancer is the risk factor of CPI-related hepatic adverse events.

[The efficacy and safety of tiotropium/olodaterol fixed-dose combination in Chinese patients with chronic obstructive pulmonary disease: a pooled subgroup analysis of TONADO 1+2].

Objective: To compare the efficacy and safety profiles of tiotropium/olodaterol with the mono-components in Chinese and total study population from TONADO trial. Methods: In the replicate, double-blind, parallel-group, active-controlled, randomized, 52-week, Phase Ⅲ TONADO studies (TONADO 1+2), patients received tiotropium/olodaterol, tiotropium, or olodaterol via the Respimat(®) Inhaler (Boehringer Ingelheim, Germany). Primary end points were forced expiratory volume in 1 second (FEV(1)) area under the curve from 0 to 3 hours (AUC(0-3h)) response and trough FEV(1) response, and St George's respiratory questionnaire (SGRQ) total score at 24 weeks. Adverse events were also collected. This subgroup analysis only focused on the efficacy and safety of the drug at the approved dose in China. Results: 548 Chinese patients were randomized, aged 41 to 82 years [mean age, (63±8) years] and most were male (526, 96%), 111 received tiotropium/olodaterol 5/5 µg, and 127 received tiotropium 5 µg and 95 received olodaterol 5 µg. The baseline characteristics of these groups were similar. After 24 weeks, treatment with tiotropium/olodaterol 5/5 µg, tiotropium 5 µg and olodaterol 5 µg resulted in an adjusted mean FEV(1) AUC(0-3h) response of 0.240, 0.157 and 0.079 L, and trough FEV(1) response of 0.117, 0.068 and-0.001 L, respectively. Tiotropium/olodaterol 5/5 µg significantly improved SGRQ scores in Chinese patients compared with olodaterol 5 µg (32.729 and 37.202, respectively). Generally, the safety profile of tiotropium/olodaterol was comparable with mono-components in 52 weeks. Conclusion: Compared with tiotropium or olodaterol, tiotropium/olodaterol in Chinese patients provided significant improvement in lung function and quality of life, and the safety profiles were similar.

[Discussion on pT3 staging in TNM staging of AJCC 8(th) edition gallbladder carcinoma].

Objective: To discuss the rationality of stage pT3 in the AJCC 8(th) TNM criteria of gallbladder carcinoma. Methods: A retrospective study was performed to analyze the clinical and pathological data of 88 patients with pT3 gallbladder carcinoma admitted to Department of Second Biliary Surgery of Eastern Hepatobiliary Surgery Hospital, affiliated to Naval Medical University from May 2013 to September 2018.pT3 stage tumors were divided into two groups: (1) pT3a stage: tumors had penetrated serosa but not directly invaded liver and/or an adjacent organ or structure; (2) pT3b stage: tumor penetrating serosa and directly invaded liver and/or an adjacent organ or structure. There were 45 patients with pT3a stage, including 15 males and 30 females, aged 36 to 80 years, with a median age of 59 years; 43 patients with pT3b, including 24 males and 19 females, aged 41 to 78 years old, median aged 63 years old.Patients with pT3a and pT3b were further divided into two groups respectively: radical resection group and extended radical resection group according to surgical radicalization. Independent sample t-test was used for comparison between two groups with normal distribution measurement data. Wilcoxon rank sum test was used between groups of non-normally distributed measurement data.The comparison of the count data was performed by χ(2) test or Fisher exact probability method. Survival analysis was performed using Kaplan-Meier method, and survival rate was compared using Log-rank test. Results: (1)Serum total bilirubin(15.6(90.3)mmol/L), albumin(40.2(4.8)mmol/L), and CA19-9(132.90(455.78)U/ml) levels in pT3b patients were higher than that in pT3a patients(10.2(6.8)mmol/L, 41.8(4.9)mmol/L, 14.35(36.27)U/ml), respectively(Z=-3.816, -1.966, -3.739, all P<0.05),postoperative complication rate in pT3b patients(24.4%) was higher than that in pT3a patients(8.9%)(P<0.05),postoperative hospital stay(12(7)days) and overall hospital stay((26±17)days) of pT3b patients were longer than that of pT3a patients((10±5) days and (19±7)days) (P<0.05). (2) The 1-, 3-, 5-year survival rates of pT3b and pT3a patients were 53%,22%,22% and 69%, 46%,38%,and the median survival time was 13 months and 26 months, respectively. The difference in survival rates between the two groups was statistically significant(χ(2)=5.117, P=0.024). (3)The 1-, 3-year survival rates of extended radical resection group(n=19) and radical resection group(n=24) in the pT3b stage were 73%, 36% and 28%, 7%, respectively.The survival time was 20 months and 9 months,respectively,and the difference in survival rates between the two groups was statistically significant(χ(2)=4.976, P=0.026). Conclusions: pT3 gallbladder carcinoma could be further subdivided into pT3a stage and pT3b stage based on the TNM criteria of AJCC 8(th) gallbladder carcinoma. Extended radical resection for pT3b gallbladder carcinoma should be further considered after comprehensive assessment of the patient's basic condition and surgical tolerance.

Improving the effectiveness of lifestyle interventions for gestational diabetes prevention: a meta-analysis and meta-regression.

BACKGROUND: Diet and exercise during pregnancy have been used to prevent gestational diabetes mellitus (GDM) with some success. OBJECTIVE: To examine the effectiveness of lifestyle intervention on GDM prevention and to identify key effectiveness moderators to improve the prevention strategy. SEARCH STRATEGY: Pubmed, Scopus, Cochrane, and cross-references were searched. SELECTION CRITERIA: Randomised controlled trials (RCTs) evaluating lifestyle interventions during pregnancy for GDM prevention. DATA COLLECTION AND ANALYSIS: Two independent reviewers extracted data. A random-effects model was used to analyse the relative risk (RR) and 95% confidence interval (95% CI). Meta-regressions and subgroup analyses were used to investigate important moderators of effectiveness. MAIN RESULTS: Forty-seven RCTs involving 15 745 participants showed that diet and exercise during pregnancy were preventive of GDM (RR 0.77, 95% CI 0.69-0.87). Four key aspects were identified to improve the preventive effect: targeting the high-risk population; an early initiation of the intervention; the correct intensity and frequency of exercise; and gestational weight gain management. Although 24 RCTs targeted women who were overweight or obese, body mass index (BMI) failed to predict the effectiveness of an intervention. Instead, interventions are most effective in high-incidence populations rather than simply in women who are overweight or obese. Furthermore, exercise of moderate intensity for 50-60 minutes twice a week could lead to an approximately 24% reduction in GDM. CONCLUSION: The best strategy to prevent GDM is to target the high-risk population predicted by risk evaluation models and to control the gestational weight gain of women through intensified diet and exercise modifications early in their pregnancy. TWEETABLE ABSTRACT: Four key effectiveness moderators of lifestyle interventions for GDM prevention.

[Relationship between coronary tortuosity and coronary microvascular disease].

Objective: To explore the relationship between coronary tortuosity and coronary microvascular disease (CMVD). Methods: Patients with typical angina symptoms and without serious coronary artery stenosis by coronary angiography were enrolled from June 2014 to December 2016, and CMVD was diagnosed by single photon emission tomography (SPECT). According to the SPECT results, patients were divided to the CMVD group and non-CMVD group. The baseline clinical characteristics, results of coronary angiography were compared between the two groups. The logistic analysis was used to analyze the relationship between coronary tortuosity and CMVD. Result: A total of 117 cases were enrolled, with 69 cases in the CMVD group and 48 cases in the non-CMVD group. No differences were found in gender distribution, age, hypertension, lipid abnormality, hyperuricemia and uses of statins between the two groups (all P>0.05). Incidence of diabetes (78.26%(54/69) vs. 35.42% (17/48) , P<0.05), hs-CRP ((4.29±2.15)mmol/L vs. (2.63±1.20)mmol/L, P<0.001), LDL-C ((2.98±0.96)mmol/L vs. (2.52±0.83)mmol/L, P=0.008) and homocysteine ((13.7±5.61)mmol/L vs. (11.5±4.38)mmol/L, P=0.025) levels were higher in the CMVD group than in the non-CMVD group. The data derived from echocardiographic examination were similar between the two groups. The Corrected TIMI frame counts were higher in the CMVD group than in non-CMVD group (LAD: 31.56±4.92 vs. 27.31±3.75, LCX: 29.47±4.18 vs. 26.62±3.19, RCA: 29.09±5.05 vs. 26.24±3.28, all P<0.001). The incidences of coronary atherosclerosis (76.81% (53/69) vs. 27.08% (13/48) , P<0.001) and coronary tortuosity ( (60.87% (42/69) vs. 33.33% (16/48) , P=0.035) were also higher in the CMVD group than in non-CMVD group. Logistic analysis found that coronary tortuosity (OR=6.111, 95%CI 2.707-13.794, P<0.001), diabetes (OR=6.565, 95%CI 2.883-14.948, P<0.001) and coronary atherosclerosis (OR=8.918, 95%CI 3.822-20.808, P<0.001) were independent risk factors of CMVD. Conclusion: Coronary tortuosity, diabetes and coronary atherosclerosis are related to CMVD in this patient cohort.

[Bortezomib inhibits hypoxia-induced increase of Orai-1 expression in pulmonary arterial smooth muscle cell].

Objective: In this study, a primary culture system for the rat distal pulmonary arterial smooth muscle cell (PASMC) was established to observe the effect of Bortezomib a treatment on the basal intracellular calcium concentration ([Ca(2+) ](i)), store operated calcium entry (SOCE) and Orai-1 expression in rat PASMC. Methods: We employed the primary culture method for the rat distal PASMC including the enzymatically dissociation of PASMC from the freshly isolated distal pulmonary artery and the culture of PASMC. The In Cyte system was used to measure the basal [Ca(2+) ](i) and SOCE after substantial treatment.Orai-1 protein expression in rat pulmonary artery smooth muscle was detected by Western blot. Results: Compared with Hypoxia group, the basal [Ca(2+) ](i) were significantly reduced in Hypoxia+ BTZ group(P<0.01). The basal [Ca(2+) ](i) A340/A380 ratio of Normoxia group was(1.07±0.02). The basal [Ca(2+) ](i) of Hypoxia group was(1.49±0.03); The Hypoxia+ BTZ group was(1.17±0.03). Compared with Hypoxia group, the store operated calcium entry were significantly reduced in Hypoxia+ BTZ group(P<0.01). The SOCE A340/A380 ratio of Normoxia group was(0.56±0.02). The SOCE of Hypoxia group was(0.84±0.02); The Hypoxia+ BTZ group was(0.66±0.02). The level of Orail-1 protein in pulmonary artery smooth muscle of Hypoxia group was (181.5±12.7)% higher than control group which was(100±0)%, (P<0.05). In the Hypoxia+ BTZ group Orai-1 protein expression was recovered(146.7±15.1)%, (P<0.05). Conclusion: Bortezomib inhibit chronically hypoxic enhancement of Orail-1 protein expression, basal [Ca(2+) ](i) and SOCE in rat distal pulmonary arterial smooth muscle cells.

[Effects of different sulfonylureas on the warm-up phenomenon in diabetes patients with coronary artery disease].

Objective: To explore the different effects of chronic treatment with glibenclamide and gliclazide on the warm-up phenomenon in diabetes patients with coronary artery disease. Methods: A total of seventy-one patients with chronic stable angina and diabetes who were positive for exercise test and was proven that the stenosis degree was 70%-90% in at least one major branch through coronary angiogram were included into the study.They were divided into three groups, diet control group (DMD), glibenclamide group (DMG1) and gliclazide group (DMG2), according to the treatment of diabetes.All of the patients underwent two bicycle exercise tests (EX) at 15-minute interval.Parameters including ischaemic threshold (the rate-pressure product at 1-mm ST-segment depression, RPP), time to ischaemic (the time to 1 mm ST-segment depression, T-STD), the maximum ST-segment depression (STDmax) and exercise duration (ED) were recorded respectively. Results: In group DMD, T-STD and ED were prolonged [(360±83) s vs (409±80) s, P<0.001] and [(518±90) s vs (549±96) s, P=0.001], STDmax were shortened [(1.91±0.43) mm vs (1.60±0.36) mm, P<0.001], and RPP was increased [(180±27) beats·min(-1)·mmHg·10(2) vs (195±28) beats·min(-1)·mmHg·10(2), P<0.001] as the parameters during EX2 were compared with those during EX1. In group DMG1, there was no statistic difference in these indexes except that ED was prolonged [(458±70) s vs (472±66) s, P=0.045] when those of EX2 and EX1were compared. In the group DMG2, all the analyzed variables improved significantly during two sequential exercise tests as the results in the group DMD except that ischaemic threshold was not increased [(199±41) beats·min(-1)·mmHg·10(2) vs (211±39) beats·min(-1)·mmHg·10(2), P=0.071]. Conclutions: Warm-up phenomenon is abolished in diabetic patients with stable angina treated with glibenclamide, partially preserved in gliclazide-treated patients. and the KATP channel may be involved in those different effects. Gliclazide should be the safer choice for the patients with diabetes and chronic stable angina.

Correlation of androgen receptor and SRD5A2 gene mutations with pediatric hypospadias in 46, XY DSD children.

We performed an exploratory study by analyzing the correlation of 46, XY disorders of sex development (46, XY DSD) with androgen receptor (AR) and steroid 5α-reductase-2 (SRD5A2) gene mutations and a safety analysis of dihydrotestosterone (DHT) gel treatment for pediatric micropenis. We collected samples from 76 pediatric patients with 46, XY DSD and 50 healthy adult men with normal fertility as the control group. The pediatric patients were treated with DHT gel (0.1-0.3 mg/kg/day) for three to six months. The extended penis length, testicular volume, and multiple blood parameters were collected before treatment and one, three, and six months after treatment. Of the 76 cases with 46, XY DSD, 31.58% had hypospadias with micropenis and 6.58% had male pseudohermaphroditism. Through AR gene screening, it was found that 14 patients had AR point mutations and 22 patients had SRD5A2 mutations. After treatment with DHT, the penis length of the patients significantly improved after one, three, and six months of treatment, with longer treatment times resulting in greater improvement. Before treatment with DHT, the average serum DHT value of patients with 46, XY DSD was 24.29 pg/mL. After one, three, and six months of treatment, this value increased to 430.71, 328.9, and 323.6 pg/mL, respectively. We conclude that for pediatric patients who have male hermaphroditism or hypospadias with micropenis, AR and SRD5A2 gene mutation detection should be performed. Local application of DHT gel can promote penis growth effectively without systemic adverse reactions.

Detection of CK19, LUNX, and KS1/4 mRNA expression in the peripheral blood for diagnosis of micrometastases in patients with non-small cell lung cancer and their clinical implications.

The expression of CK19, LUNX, and KS1/4 mRNA biomarkers was detected in the peripheral blood of non-small cell lung cancer (NSCLC) patients to investigate the feasibility of indicating lung cancer micrometastases. Micrometastases were identified in the peripheral blood of 32 NSCLC patients, 15 benign pulmonary disease (BPD) patients, and 10 healthy volunteers by reverse transcriptase-polymerase chain reaction. The detection rates of CK19, LUNX, and KS1/4 mRNA-positive cells in the peripheral blood obtained from the NSCLC group were 34.4% (11/32), 37.5% (12/32), and 25% (8/32), respectively. CK19, LUNX, and KS1/4 mRNA-positive cells were detected in 6.6% (1/15), 0.0% (0/15), and 13.3% (2/15) of the patients with BPD, respectively. However, the healthy group did not express any of the three markers. The expression of CK19, LUNX, and KS1/4 mRNA was significantly higher in the NSCLC group than that in the healthy and BPD groups (P < 0.05). CK19 and LUNX mRNA may be ideal biomarkers indicating micrometastases in patients with NSCLC; however, the diagnostic applicability of KS1/4 mRNA remains uncertain. The rate of expression of CK19 was not correlated with the clinicopathological characteristics (P > 0.05). The rate of expression of LUNX and KS1/4 was closely related to the clinical stage (P < 0.05), and not related to the clinical characteristics of the disease (age, gender, smoking history, pathological type, histologic classification, and differentiation; P > 0.05).

Genetic diversity analysis of eight indigenous goat breeds (groups) in China using AFLP markers.

Eight goat breeds from Chengdu, Lezhi, Jintang, Baiyu, Yingshan, Hejiang, Jiangan, and Jialing in Sichuan, China were analyzed for genetic diversity using AFLP markers. A total of 261 individuals were analyzed using eight selective AFLP primer combinations which generated 174 discernible fragments, of which 80 were polymorphic with an average of 27.15 bands per primer pair. Shannon's gene diversity index among breeds ranged from 0.0888 to 0.2289, and the highest and the lowest gene diversity index among breeds were the Yingshan Black goat (0.2289) and the Baiyu Black goat (0.0888) respectively. The genetic distance (DR) among eight goat breeds were calculated by Rogers' genetic distance. Based on the analytical results, systematic trees were constructed by using the unweighted pair-group method with arithmetic average (UPGMA) and the neighbor joining (NJ) methods. Eight breeds were divided into two main clusters by using cluster analysis, one of which including two subclusters. The clustering results agreed well with the origin of the goats and geographic distribution. The results implied that AFLP markers were informative and could provide significant insights for genetic diversity research and genetic resource conservation of indigenous goats.

Determination of mizoribine in human plasma using high-performance liquid chromatography: application to a pharmacokinetic study in Chinese renal transplant recipients.

A rapid, simple, and reproducible high-performance liquid chromatographic method (HPLC) was developed and validated for the determination of mizoribine in human plasma using cytarabine as internal standard (IS). The plasma samples of mizoribine were precipitated with 6% perchloric acid. The supernatant was separated on a reversed phase C18 column with a mobile phase of 10 mM KH2PO4 buffer solution (pH 6.3) containing 10 mM perchloric acid using isocratic elution (at flow rate 1.5 mL/min), and detected using an ultraviolet detector at 280 nm. The assay exhibited a linear range of 0.02-10.0 µg/mL for mizoribine in human plasma and the lower limit of quantification was 0.02 µg/mL. The method was statistically validated for linearity, accuracy, precision and selectivity. In addition, the method was successfully applied to estimate the pharmacokinetic parameters of mizoribine in Chinese kidney transplant patients following an oral administration of 100 mg mizoribine (2 Bredinin® 50 mg tablets).

Epithelial-mesenchymal transition biomarkers and support vector machine guided model in preoperatively predicting regional lymph node metastasis for rectal cancer.

BACKGROUND: Current imaging modalities are inadequate in preoperatively predicting regional lymph node metastasis (RLNM) status in rectal cancer (RC). Here, we designed support vector machine (SVM) model to address this issue by integrating epithelial-mesenchymal-transition (EMT)-related biomarkers along with clinicopathological variables. METHODS: Using tissue microarrays and immunohistochemistry, the EMT-related biomarkers expression was measured in 193 RC patients. Of which, 74 patients were assigned to the training set to select the robust variables for designing SVM model. The SVM model predictive value was validated in the testing set (119 patients). RESULTS: In training set, eight variables, including six EMT-related biomarkers and two clinicopathological variables, were selected to devise SVM model. In testing set, we identified 63 patients with high risk to RLNM and 56 patients with low risk. The sensitivity, specificity and overall accuracy of SVM in predicting RLNM were 68.3%, 81.1% and 72.3%, respectively. Importantly, multivariate logistic regression analysis showed that SVM model was indeed an independent predictor of RLNM status (odds ratio, 11.536; 95% confidence interval, 4.113-32.361; P<0.0001). CONCLUSION: Our SVM-based model displayed moderately strong predictive power in defining the RLNM status in RC patients, providing an important approach to select RLNM high-risk subgroup for neoadjuvant chemoradiotherapy.

[A comparative study on the patients with VVI and AAI pacing by equilibrium radionuclide angiography].

Twenty two patients (15 cases of VVI and 7 cases of AAI) were studied before and after implanted pacemaker comparatively with equilibrium radionuclide angiography (ERNA). The results of the phase analysis (PA) and the cardiac function parameters showed that the ventricular excitement conduction-systolic sequence (VEC-SS) of VVI pacemaker group was abnormal, the ventricular systolic coordination (VSC) and the global ventricular systolic function (GVSF) were reduced, while the early ventricular ejection fraction and the diastolic filling velocity were increased after implantation VEC-SS and VSC of AAI group showed no change and the GVSF got improved after implantation. VES-SS of VVI group was abnormal, VSC and GVSF of VVI group were reduced as compared with those of control group, but no changes were found in AAI group after implantation. Hemodynamics appeared to be better in AAI group than that of VVI group, as evidenced by normal VEC-SS and VSC as well as increased GVSE.

[Studies on hyperglycophilic yeast causing gaseous fermentation of sweetened condensed milk].

This report studied on a hyperglycophilic yeast isolated from sweetened condensed milk in Shandong province. It can fermented glucose, sucrose and raffinose; and can assimilate glucose, sucrose and raffinose as its sole carbon source. According to the morphology, physiology and habitat, it was identified to be Candida lactiscondensi (Hammer) Meyer et Yarrow, one of the synonym is Torulopsis lactiscondensi (Hammer) Lodder et Kreger-van Rij (1952). Detailed description was given in this report.