PTBP1 promotes the progression of hepatocellular carcinoma by enhancing the oncogenic splicing switch of FGFR2.

Hepatocellular carcinoma (HCC) is the most common type of primary liver cancer accounting for 90% of cases. It is a highly invasive and deadly cancer with a gradual onset. Polypyrimidine tract-binding protein 1 (PTBP1) is an important RNA-binding protein involved in RNA metabolism and has been linked to oncogenic splicing events. While the oncogenic role of PTBP1 in HCC cells has been established, the exact mechanism of action remains unclear. This study aimed to investigate the functional connection between PTBP1 and dysregulated splicing events in HCC. Through immunoprecipitation-mass spectrometry analyses, we discovered that the proteins bound to PTBP1 were significantly enriched in the complex responsible for the alternative splicing of FGFR2 (fibroblast growth factor receptor 2). Further RNA immunoprecipitation and quantitative PCR assays confirmed that PTBP1 down-regulated the FGFR2-IIIb isoform levels and up-regulated the FGFR2-IIIc isoform levels in HCC cells, leading to a switch from FGFR2-IIIb to FGFR2-IIIc isoforms. Subsequent functional evaluations using CCK-8, transwell, and plate clone formation assays in HCC cell lines HepG2 and Huh7 demonstrated that FGFR2-IIIb exhibited tumor-suppressive effects, while FGFR2-IIIc displayed tumor-promoting effects. In conclusion, this study provides insights into the PTBP1-mediated alternative splicing mechanism in HCC progression, offering a new theoretical basis for the prevention and treatment of this malignancy. Mechanistically, the isoform switch from FGFR2-IIIb to FGFR2-IIIc promoted epithelial-mesenchymal transformation (EMT) of HCC cells and activated the FGFR cascades ERK and AKT pathways.

Development of an in vitro screening method for idiosyncratic hepatotoxic components in traditional Chinese medicine: a case study with Epimedii Folium and Psoraleae Fructus / 药学学报

Idiosyncratic drug-induced liver injury (IDILI) has long posed a challenging and pivotal concern in pharmaceutical research. The complex composition of traditional Chinese medicine (TCM) has introduced a bottleneck in current research, hindering the elucidation of the component basis associated with IDILI in TCM. Using Epimedii Folium (EF) and Psoraleae Fructus (PF) as illustrative examples, this study endeavors to establish an in vitro evaluation model, providing a high-throughput and preliminary assessment method for screening components related to TCM-induced IDILI. A TNF-α-mediated HepG2 susceptible model was first established in this study, with the focus on the index components present in EF and PF. The release of lactate dehydrogenase (LDH) in the cell supernatant served as the detection index. A concentration-toxicity response curve was constructed, and the hepatotoxic components of EF and PF were identified utilizing the synergistic toxicity index. The LDH results unveiled the hepatotoxic effects of bavachin, backuchiol, isobavachin, neobavaisoflavone, psoralidin, isobavachalcone, icarisid I, and icarisid II on both normal and susceptible cells, categorizing these 8 components as both direct hepatotoxicity components and idiosyncratic hepatotoxicity components. Bavachin and neobavaisoflavone exhibited no hepatotoxicity on normal cells but demonstrated significant effects on susceptible cells, designating them as potential idiosyncratic susceptible hepatotoxicity components. The study further delineated that 10 EF components and 3 PF components were direct immune-promoting hepatotoxicity components. Additionally, 14 idiosyncratic immune-promoting hepatotoxicity components were identified, encompassing 10 EF components and 4 PF components, with neobavaisoflavone, bavachinin, and isobavachin being potential idiosyncratic susceptible immune-promoting hepatotoxicity components. Synergistic toxicity index results indicated that 13 idiosyncratic immune-promoting hepatotoxicity components (except anhydroicaritin) combined with bavachin demonstrated synergistic hepatotoxicity on susceptible cells. Notably, 3 idiosyncratic susceptible immune-promoting hepatotoxicity components combined with bavachin exhibited synergistic hepatotoxicity, with neobavaisoflavone displaying the highest synergistic toxicity index and bavachinin the lowest. In summary, this methodology successfully screens hepatotoxic and immune-promoting hepatotoxic components in EF and PF, distinguishing the types of components inducing hepatotoxicity, evaluating the hepatotoxicity degree of each component, and elucidating the synergistic relationships among them. Importantly, these findings align with the characteristics of IDILI. The method provides an effective model tool for the fundamental research of TCM-related IDILI components.

Research on equity of medical resource allocation in Yangtze River Economic Belt under healthy China strategy.

Objective: This study aimed to assess the fairness of medical resource allocation in the Yangtze River Economic Belt, based on the Healthy China strategy. It aimed to identify the issues with resource allocation fairness and provide optimization suggestions. Methods: To assess the allocation fairness from a geographical population perspective, the study used the Health Resource Concentration and Entropy Weight TOPSIS methods. Additionally, the study analyzed the allocation fairness from an economic level angle, using the Concentration Curve and Concentration Index. Results: The study found that the downstream area had higher resource allocation fairness than the midstream and upstream areas. The middle reaches had more resources than the upper and lower reaches, based on population concentration. The Entropy Weight TOPSIS method found that Shanghai, Zhejiang, Chongqing, and Jiangsu had the highest comprehensive score index of agglomeration. Furthermore, from 2013 to 2019, the fairness of medical resource distribution gradually improved for different economic levels. Government health expenditure and medical beds were distributed more equitably, while general practitioners had the highest level of unfairness. However, except for medical and health institutions, traditional Chinese medicine institutions, and primary health institutions, other medical resources were mostly distributed to areas with better economic conditions. Conclusion: The study found that the fairness of medical resource allocation in the Yangtze River Economic Belt varied greatly based on geographical population distribution, with inadequate spatial accessibility and service accessibility. Although the fairness of distribution based on economic levels improved over time, medical resources were still concentrated in better economic areas. The study recommends improving regional coordinated development to enhance the fairness of medical resource allocation in the Yangtze River Economic Belt.

Clinical effect and follow-up of laparoscopic radical proximal gastrectomy for upper gastric carcinoma.

Objective: To evaluate the safety and clinical effect of tubular esophagogastric anastomosis in laparoscopic radical proximal gastrectomy. Methods: A retrospective analysis was conducted involving 191 patients who underwent laparoscopic radical proximal gastrectomy in the Department of Gastrointestinal Surgery, Qilu Hospital of Shandong University from January 2017 to October 2020. Patients were divided into tubular esophagogastric anastomosis group (TG group) and traditional esophagogastric anastomosis group (EG group) according to the digestive tract reconstruction. Their intraoperative conditions, perioperative recovery and postoperative follow-up were compared. Patients were also divided into indocyanine green group and non-indocyanine green group according to whether or not indocyanine green tracer technology was used during the operation. Their intraoperative condition and perioperative recovery were compared and analyzed after propensity score matching. Results: The operation was successfully completed in all patients. Compared with the EG group, the TG group had less volume of gastric tube drainage, shorter gastric tube drainage time and proton pump inhibitors application time, and lower reuse rate of proton pump inhibitors. However, the TG group had a higher anastomotic stenosis at three months after surgery, as measured using anastomotic width and dysphagia score. Nevertheless, the incidence of reflux esophagitis and postoperative quality of life score in the TG group were lower compared with the EG group at 1st and 2nd year after surgery. In the indocyanine green analysis, the indocyanine green group had significantly shorter total operation time and lymph node dissection time and less intraoperative blood loss compared with the non-indocyanine green group. However, compared with the non-indocyanine green group, more postoperative lymph nodes were obtained in the indocyanine green group. Conclusion: Laparoscopic radical proximal gastrectomy is safe and effective treatment option for upper gastric cancer. Tubular esophagogastric anastomosis has more advantages in restoring postoperative gastrointestinal function and reducing reflux, but it has a higher incidence of postoperative anastomotic stenosis compared with traditional esophagogastrostomy. The application of indocyanine green tracer technique in laparoscopic radical proximal gastrectomy has positive significance.

Sedentary behavior, screen time and mental health of college students: a Meta-analysis / 中华流行病学杂志

Objective: To evaluate the effects of sedentary behavior/screen time on mental health of college students by Meta-analysis based on the results of literature retrieval and provide theoretical basis for the improvement of college students' mental health. Methods: The original research literatures about sedentary behavior (including screen time) and college students' mental health published as of 14 July 2022 were retrieved from PubMed, Embase, Cochrane Library, CNKI, VIP and Wanfang data. Data were extracted from the included studies and scored by one author in accordance with the proposed programme, and quality score was reviewed by another author. The literature that met the inclusion criteria was systematically reviewed and Meta-analysis was carried out by software Stata 14.2 based on the data from the literatures. Results: A total of 36 studies met the inclusion criteria, including 35 observational studies and 1 interventional study. There are 4 papers about the effects of sedentary behavior and 9 papers about the effects of screening time on depression in college students and 4 papers about the effects of sedentary behavior/screening time on anxiety in college students were used for a Meta-analysis, and the other studies were also analyzed. The Meta-analysis on the effects of sedentary behavior on depression in college students showed that there was a significant positive correlation between higher level sedentary behavior and increased risk for depression (OR=1.07,95%CI:1.05-1.10). Subgroup analysis indicated that there was no significant correlation between higher level sedentary behavior and depression (OR=1.74, 95%CI:0.93-3.25) in the unadjusted confounding factor model, but there was significance positive correlation after adjusting confounding factors (OR=2.15, 95%CI:1.18-3.90). Meta-analysis on the effects of screen time on depression in college students showed that longer screen time were significantly positively correlated with higher depression level (OR=1.03, 95%CI: 1.02-1.05). The results of subgroup analysis showed that in both unadjusted confounding factor model and adjusted confounding factor model, longer screen time was significantly positively correlated with depression (OR=1.27, 95%CI: 1.13-1.42; OR=1.45, 95%CI: 1.18-1.79) , respectively. Meta-analysis on the effects of sedentary behavior on anxiety showed that longer screen time was significantly positively correlated with increased anxiety risk (OR=1.44, 95%CI: 1.31-1.58). The results of subgroup analysis showed that in both unadjusted confounding factor model and adjusted confounding factor model, there was a significant positive correlation between longer screen time and anxiety (OR=1.47, 95%CI: 1.31-1.65; OR=1.38, 95%CI:1.17-1.62). The analysis for the literatures which were not eligible for Meta-analysis found that sedentary behavior/screen time was significantly associated with stress and other mental health in college students. Conclusions: Sedentary behavior or screen time is significantly negatively correlated with college students' mental health, in particular, resulting in depression and anxiety. These effects might be be different between weekdays and weekend days.

PTBP1 Promotes Growth, Migration and Invasion of Cholangiocarcinoma Cells by Regulating mRNA Alternative Splicing / 中国生物化学与分子生物学报

Cholangiocarcinoma (CCA) is a highly invasive type of cancer with insidious onset and high mortality. Polypyrimidine tract-binding protein 1 (PTBP1) is highly over-expressed in various types of tumor tissues, which contributes to cancer progression. But the role of PTBP1 in CCA has not been explored yet. In this study, we aim to investigate the function of PTBP1 in CCA. Therefore, we used publicly available data from the cancer genome atlas (TCGA) to evaluate the dysregulation of PTBP1 in CCA. The results showed that the PTBP1 is significantly up-regulated in CCA tissues compared to the matched non-tumor tissues (P < 0. 05). We assessed the effects of PTBP1 on the growth of CCA cell lines RBE and HuH28 by performing CCK-8 and plate colony formation assays. The results showed that overexpression of PTBP1 significantly promoted the growth (P < 0. 01) of CCA cells, whereas knockdown of PTBP1 exhibited opposite effects. Transwell and Invasion assays revealed that overexpression of PTBP1 significantly promotes the migration and invasion of CCA cells (P < 0. 001), whereas knockdown of PTBP1 exhibited opposite effects (P < 0. 001). The RNA sequencing (RNA-seq) analysis in PTBP1-depleted cells showed that the up-regulated genes are significantly enriched in p53 signaling pathway, while the down-regulated genes are represented by cholesterol metabolism, Rho GTPase and TGF-β pathways. Then, the alternative splicing analysis revealed that inhibition of PTBP1 led to series of aberrant alternative splicing events, including several cancer-associated ones, such as splicing events within the TGF-β regulator TGIF1 and the p53 activity-correlated gene GNAS. These results indicate that PTBP1 promotes the progression of CCA likely by regulating the transcriptome alternative splicing to influence multiple cancer-associated signaling pathways.

A nomogram for predicting lymph node metastasis in early gastric cancer / 中华胃肠外科杂志

Objective: To explore the independent risk factors of lymph node metastasis (LNM) in early gastric cancer, and to use nomogram to construct a prediction model for above LNM. Methods: A retrospective cohort study was conducted. Inclusion criteria: (1) primary early gastric cancer as stage pT1 confirmed by postoperative pathology; (2) complete clinicopathological data. Exclusion criteria: (1) patients with advanced gastric cancer, stump gastric cancer or history of gastrectomy; (2) early gastric cancer patients confirmed by pathology after neoadjuvant chemotherapy; (3) other types of gastric tumors, such as lymphoma, neuroendocrine tumor, stromal tumor, etc.; (4) primary tumors of other organs with gastric metastasis. According to the above criteria, 1633 patients with early gastric cancer who underwent radical gastrectomy at the Department of General Surgery of the Chinese PLA General Hospital First Medical Center from December 2005 to December 2020 were enrolled as training set, meanwhile 239 patients with early gastric cancer who underwent gastrectomy at the Department of General Surgery of the Chinese PLA General Hospital Fourth Medical Center from December 2015 to December 2020 were enrolled as external validation set. Risk factors of LNM in early gastric cancer were identified by using univariate and multivariate logistic regression analyses. A nomogram prediction model was established with significant factors screened by multivariate analysis. Area under the receiver operating characteristic curve (AUC) was used for assessing the predictive value of the model. Calibration curve was drawn for external validation. Results: Among 1633 patients in training set, the mean number of retrieved lymph nodes was 20 (13-28), and 209 patients (12.8%) had lymph node metastasis. Univariate analysis showed that gender, resection range, tumor location, tumor morphology, lymph node clearance, vascular invasion, lymphatic cancer thrombus, tumor length, tumor differentiation, microscopic presence of signet ring cells and depth of tumor invasion were associated with LNM (all P<0.05). Multivariate analysis revealed that females, tumor morphology as ulcer type, vascular invasion, lymphatic cancer thrombus, tumor length≥3 cm, deeper invasion of mucosa, and poor differentiation were independent risk factors for LNM in early gastric cancers (all P<0.05). Receiver operating characteristic curve indicated that AUC of training set was 0.818 (95%CI: 0.790-0.847) and AUC of external validation set was 0.765 (95%CI: 0.688-0.843). The calibration curve showed that the LNM probability predicted by nomogram was consistent with the actual situation (C-index: 0.818 in training set and 0.765 in external validation set). Conclusions: Females, tumor morphology as ulcer type, vascular invasion, lymphatic cancer thrombus, tumor length≥3 cm, deeper invasion of mucosa and poor differentiation are independent risk factors for LNM of early gastric cancer. The establishment of a nomogram prediction model for LNM in early gastric cancer has great diagnostic value and can provide reference for treatment selection.

Analysis of influence factors for short-term recurrence of retroperitoneal liposarcoma after complete resection / 中华外科杂志

Objective: To examine the influence factors of short-term recurrence after complete surgical resection of retroperitoneal liposarcoma. Methods: The clinicopathological data of retroperitoneal liposarcoma at Department of General Surgery, the First Medical Center, People's Liberation Army General Hospital from January 2000 to January 2020 were retrospectively analyzed. There were 60 males and 31 females, aged (52.1±9.9) years (range: 30 to 84 years). Tumor recurrence within 12 months after complete resection was defined as short-term recurrence, and tumor recurrence more than 12 months was defined as non-short-term recurrence. The t test, rank-sum test, χ2 test and Fisher exact test were conducted for inter-group comparison. Logistic regression analysis was used to analyze the independent influence factors for the short-term recurrence of retroperitoneal liposarcoma after complete resection. The Kaplan-Meier curve was used to calculate the recurrence-free survival, and the Log-rank test was adopted for the comparison between the groups. Results: The univariate analysis results showed that irregular tumor morphology, multiple pathological subtypes, pathological scores>3, and multiple primary tumors are influence factors for short-term recurrence after complete resection of retroperitoneal liposarcoma (χ2: 4.422 to 7.773, all P<0.05). Regression analysis of the above risk factors showed that multiple primary tumors was the independent risk factor (OR=2.918, 95%CI: 1.127 to 7.556, P=0.027). In the short-term recurrence group, Kaplan-Meier curve analysis showed that patients with multiple primary tumors had a shorter median recurrence time than patients with unifocal tumor (6 months vs. 9 months, P=0.028). Conclusions: Multiple primary tumor is an independent risk factor for short-term recurrence after complete resection of retroperitoneal liposarcoma. It suggests that the frequency of follow-up after surgery should be increased for such patients.

Effect of Curcumin on Apoptosis of Acute T-Lymphoblastic Leukemia Cells induced by UMI-77 and Its Related Mechanism / 中国实验血液学杂志

AbstractObjective: To explore the effect and mechanism of curcumin on human T-cell acute lymphoblastic leukemia (T-ALL) cell apoptosis induced by Mcl-1 small molecule inhibitors UMI-77.@*METHODS@#T-ALL cell line Molt-4 was cultured, and the cells were treated with different concentrations of curcumin and Mcl-1 small molecule inhibitor UMI-77 for 24 h. The MTT method was used to detect the cell survival rate after different treatment; According to the results of curcumin and UMI-77, the experimental settings were divided into control group, curcumin group (20 μmol/L curcumin treated cells), UMI-77 group (15 μmol/L Mcl-1 small molecule inhibitor UMI-77 treated cells) and curcumin+ UMI-77 group (20 μmol/L curcumin and 15 μmol/L Mcl-1 small molecule inhibitor UMI-77 treated cells), MTT method was used to detect cell proliferation inhibition rate, Annexin V-FITC/PI double staining method and TUNEL staining were used to detect cell apoptosis, DCFH-DA probe was used to detect cell reactive oxygen species, JC-1 fluorescent probe was used to detect mitochondrial membrane potential, Western blot was used to detect the expression levels of apoptosis-related proteins and Notch1 signaling pathway-related proteins.@*RESULTS@#After the treatment of Molt-4 cells with different concentrations of curcumin and Mcl-1 small molecule inhibitor UMI-77, the cell survival rate was decreased (P<0.05); Compared with the control group, the cell proliferation inhibition rate of the curcumin group and the UMI-77 group were increased, the apoptosis rate of cell was increased, the level of ROS was increased, the protein expression of Bax, Caspase-3 and Caspase-9 in the cells were all increased, and the protein expression of Bcl-2 was reduced (P<0.05); Compared with the curcumin group or UMI-77 group, the cell proliferation inhibition rate and apoptosis rate of the curcumin+UMI-77 group were further increased, and the level of ROS was increased. At the same time, the protein expression of Bax, Caspase-3 and Caspase-9 in the cells were all increased, the protein expression of Bcl-2 was reduced (P<0.05); In addition, the mitochondrial membrane potential of the cells after curcumin treatment was decreased, and the proteins expression of Notch1 and HES1 were reduced (P<0.05).@*CONCLUSION@#Curcumin can enhance the apoptosis of T-ALL cells induced by Mcl-1 small molecule inhibitor UMI-77 by reducing the mitochondrial membrane potential, the mechanism may be related to the inhibition of Notch1 signaling pathway.

Rapid chemome profiling of Cistanche salsa using DI-MS/MS~(ALL) / 中国中药杂志

The current study aims to rapidly and comprehensively profile the chemical composition of Cistanche salsa using direct infusion coupled with MS/MS~(ALL)(DI-MS/MS~(ALL)). The C. salsa extract was directly imported into electrospray ionization(ESI) source of quadrupole time-of-flight(Q-TOF) mass spectrometer with an infusion pump at a flow rate of 10 μL·min~(-1). Acquisition program was applied under negative ionization polarity to collect one MS~1 spectrum(m/z 50-1 200), followed by 1 150 MS~2 spectra with precursor isolation window(m/z 1) amongst mass range m/z 50-1 200. After each MS~2 spectrum was matched to its precursor ion, putative identification was conducted through matching mass spectral data with literature and database. A total of 31 components were identified from C. salsa, including 9 phenylethanoid glycosides, 2 iridoids, 4 saccharides, 9 organic acids, and 7 other compounds, similar to those from C. tubulosa and C. deserticola. In conclusion, DI-MS/MS~(ALL), a facile and reliable analytical tool, can be employed for qualitative analysis of chemical constituents in C. salsa. The research offers a promising strategy to achieve rapid chemome profiling of herbal medicine and provides an alternative source of Cistanches Herba.

Short-term efficacy of Billroth Ⅱ+Braun anastomosis versus Roux-en-Y anastomosis in totally three-dimensional laparoscopic distal gastrectomy / 中华消化外科杂志

Objective:To compare the short-term efficacy of Billroth Ⅱ+Braun anasto-mosis versus Roux-en-Y anastomosis in totally three-dimensional (3D) laparoscopic distal gastrectomy.Methods:The retrospective cohort study was conducted. The clinicopathological data of 140 patients with gastric cancer who were admitted to the First Medical Center of Chinese PLA General Hospital from January 2016 to January 2020 were collected. There were 105 males and 35 females, aged from 23 to 84 years, with a median age of 55 years. Of the 140 patients, 54 patients undergoing totally 3D laparoscopic distal gastrectomy with Billroth Ⅱ+Braun anastomosis were allocated into Billroth Ⅱ+Braun group, and 86 patients undergoing totally 3D laparoscopic distal gastrectomy with Roux-en-Y anastomosis were allocated into Roux-en-Y group, respectively. Observation indicators: (1) surgical situations; (2) postoperative situations; (3) follow-up. Follow-up using outpatient examination and telephone interview was conducted to detect remnant gastritis and its severity, bile reflux, reflux esophagitis in the postoperative 3 months up to April 2020. Measurement data with normal distribution were represented as Mean± SD, and comparison between groups was analyzed using the t test. Count data were described as absolute numbers or percentages, and comparison between groups was analyzed using the chi-square test or Fisher exact probability. Comparison of ordinal data was analyzed using the Mann-Whitney U test. Results:(1) Surgical situations: 140 patients underwent totally 3D laparoscopic distal gastrectomy. The operation time, cases with volume of intraoperative blood loss <50 mL, 50 to 200 mL or >200 mL, the number of lymph node dissected were (233±39)minutes,15, 35, 4, 30±13 for the Billroth Ⅱ +Braun group , respectively, versus (240±52)minutes,25, 51, 10, 27±10 for the Roux-en-Y group, showing no significant difference between the two groups ( t=0.856， χ2=0.774， t=1.518， P>0.05). (2) Postoperative situations: cases with drainage tube, time to postoperative first flatus, cases with postoperative grade Ⅱ, Ⅲ, Ⅳ, Ⅴ complications, cases with postoperative complications, cases with postoperative severe complications, duration of postoperative hospital stay, surgery cost and total hospitalization cost of the Billroth Ⅱ+Braun group were 38, (3.5±0.8)days,4, 1, 0, 0, 5, 1, (9.0±5.0)days, (3.8±1.2)×10 4 yuan and (9.7±2.1)×10 4 yuan, respectively. The above indicators of the Roux-en-Y group were 59, (3.7±1.0)days, 9, 1, 0, 1, 11, 2, (9.0±4.0)days, (4.3±1.0)×10 4 yuan and (9.2±2.1)×10 4 yuan, respectively. There was a significant difference in the surgery cost between the two groups ( t=2.453, P<0.05), while there was no significant difference in cases with drainage tube, time to postoperative first flatus, cases with postoperative grade Ⅱ, Ⅲ, Ⅳ, Ⅴ complications, cases with postoperative complications, duration of postoperative hospital stay or total hospitalization cost between the two groups ( χ2=0.049, t=?1.339, Z=0.000, χ2=0.409, t=0.197, 1.383, P>0.05). There was also no significant difference in cases with postoperative severe complications between the two groups ( P>0.05).(3) Follow-up: 134 of 140 patients received the follow-up, including 52 cases in the Billroth Ⅱ+Braun group and 82 cases in the Roux-en-Y group. Results of follow-up within postoperative 3 months showed that the incidence rates of remnant gastritis, bile reflux, reflux esophagitis were 61.5%(32/52), 38.5%(20/52), 26.9%(14/52) for the Billroth Ⅱ+Braun group, respectively, versus 41.5%(34/82), 22.0%(18/82), 12.2%(10/82) for the Roux-en-Y group, showing significant differences between the two groups ( χ2=5.131, 4.270, 4.695, P<0.05). Cases with grade 0,Ⅰ,Ⅱ, Ⅲ, Ⅳ residual food were 42, 3, 5, 2,0 for the Billroth Ⅱ+Braun group, versus 67, 9, 1, 5,0 for the Roux-en-Y group, showing no significant difference between the two groups ( Z=?0.156, P>0.05). Cases with minimal lesion, grade A, grade B gastritis (severity of gastritis) were 6, 5, 3 for the Billroth Ⅱ+Braun group, versus 8, 2, 0 for the Roux-en-Y group, showing no significant difference between the two groups ( Z=?1.468, P>0.05). Conclusions:It is safe and feasible to operate Billroth Ⅱ+Braun or Roux-en-Y anastomosis in totally 3D laparoscopic distal gastrectomy. Billroth Ⅱ+Braun anastomosis can reduce the surgical cost. Roux-en-Y anastomosis has advantages in reducing the incidence of reflux esophagitis, bile reflux and reflux gastritis.

Risk factors of postoperative complication after total gastrectomy in advanced gastric cancer patients receiving neoadjuvant chemotherapy / 中华胃肠外科杂志

Objective: At present, there are few studies focusing on the factors short-term complications after total gastrectomy in patients with advanced gastric cancer receiving neoadjuvant chemotherapy (NACT). The purpose of this study is to provide a reference for clinical prevention of complications in these patients. Methods: A retrospective case-control study was conducted. Case inclusion criteria: (1) clinical stage II-III gastric cancer diagnosed by preoperative gastroscopy, pathology, abdominal CT, EUS or PET-CT; (2) evaluated suitable for NACT by MDT discussion; (3) no previous history of other malignant tumors and no concurrent tumor; (4) undergoing total gastrectomy+ D2 lymphadenectomy after NACT. Exclusion criteria: (1) age <18 or >80 years old; (2) severe concurrent diseases, and ASA classification>grade III; (3) stump gastric cancer or history of gastric surgery; (4) incomplete clinicopathological data. According to the above criteria, clinicopathological data of 140 advanced gastric cancer patients who underwent total gastrectomy after NACT in Chinese PLA General Hospital between June 2012 and June 2019 were collected, including 109 males and 31 females with mean age of (56.9±11.4) years and body mass indey (BMI) of (23.3±3.1) kg/m(2). Logistic analysis was used to analyze the relationship between postoperative complication and clinicopathological data. Factors in univariate analysis with P<0.05 were included in the multivariate analysis. Results: Postoperative complications (Clavien-Dindo classification ≥ II) occurred in 35 cases (25.0%) and severe complications (Clavien-Dindo classification ≥ IIIa) occurred in 4 cases (2.9%), including 1 case of esophago-jejunal anastomotic leakage, 1 case of vena cava thrombosis, 1 case of pleural effusion, 1 case of septic shock during perioperative days resulting in death. Univariate analysis showed that BMI (P=0.011), cycle of NACT (P=0.027), tumor diameter (P=0.021), and vascular invasion (P=0.033) were associated with postoperative complication within 30 days, while open/laparoscopic total gastrectomy were not associated with postoperative complication (P=0.926). Multivariate analysis revealed that BMI ≥ 25 kg/m(2) (OR=3.294, 95% CI: 1.343-8.079, P=0.009) and < 4 cycles of NACT (OR=2.922, 95% CI: 1.217-7.016, P=0.016) were independent risk factors for postoperative complication. The 3-year overall survival rates of patients with or without complication were 54.4% and 64.0%, respectively (P=0.395), and 3-year disease-free survival rates were 47.4% and 52.9%, respectively (P=0.587). Conclusions: Higher BMI and fewer cycles of NACT are independent risk factors of postoperative complication in advanced gastric cancer patients undergoing total gastrectomy after NACT. No obvious association is found between postoperative complication and surgical approaches.

Relationship between lateral hypothalamus and melatonin-induced reduction of wakefulness in rats and the receptor mechanism / 中华麻醉学杂志

Objective:To investigate the relationship between lateral hypothalamus and melatonin-induced reduction of wakefulness in rats and the receptor mechanism.Methods:Forty clean-grade adult male Sprague-Dawley rats, weighing 250-300 g, were divided into 4 groups ( n=10 each) using a random number table method: control group (group C), melatonin group (group M), melatonin type-1/2 receptor (MT 1R) antagonist luzindole plus melatonin group (group L+ M), and melatonin type-2 receptor (MT 2R) antagonist 4P-PDOT plus melatonin group (P+ M group). In group C, 0.5 μl of 0.9% NaCl solution was microinjected into the lateral hypothalamus.In group M, 1 μmol/L melatonin 0.5 μl was microinjected into the lateral hypothalamus.In group L+ M, 1 μmol/L MT 1/2R and 1 μmol/L melatonin (0.5 μl in total) was microinjected into the lateral hypothalamus.The microinjection time was from 19: 30 to 20: 00.The changes in sleep-wake duration and the oscillating energy in different frequency bands of electroencephalogram were detected by using electroencephalogram and electromyogram recording technology. Results:Compared with group C, the percentage of wakefulness time was significantly decreased, the percentage of non-rapid eye movement sleep and rapid eye movement sleep time was increased, the energy for delta oscillation was increased, the energy for theta oscillation was decreased, and no significant change was found in the energy for alpha oscillation in M and P+ M groups ( P<0.01), and no significant change was found in the parameters mentioned above in group L+ M ( P>0.05). Compared with group M, the percentage of wakefulness time was significantly increased, the percentage of non-rapid eye movement sleep and rapid eye movement sleep time was decreased, the energy for delta oscillation was decreased, and the energy for theta oscillation was increased in group L+ M ( P<0.01), and no significant change was found in the parameters mentioned above in group P+ M ( P>0.05). Conclusion:The lateral hypothalamus may be involved in melatonin-induced reduction of wakefulness in rats, and the mechanism may be related to activating MT 1R in the lateral hypothalamus.

Determination of seven polycyclic aromatic hydrocarbons in water by dispersive liquid-liquid microextraction based on solidification of floating organic droplet with gas chromatography-mass spectrometry / 上海预防医学

Objective To develop a new method for the simultaneous determination of seven polycyclic aromatic hydrocarbons(PAHs)in water by dispersive liquid-liquid microextraction based on solidification of floating organic droplet(DLLME-SFO)with gas chromatography-mass spectrometry(GC-MS). Methods The experimental conditions of DLLME-SFO were determined with dodecanol as extractant solvent, methanol as dispersive solvent, inonic strength increased by adding 8% NaCl. After vortexed for 1 min and centrifuged at 4 000 r/min for 5 min, the water sample was cooled down in an ice bath till dodecanol became solid and formed a small ball. Then the solidified dodecanol phase was transferred, and directly detected by GC-MS method after it melted. Results Good linearities were obtained for the seven polycyclic aromatic hydrocarbons within the range of 5 μg/L-200 μg/L. The correlation coefficients were above 0.996. The detection limits ranged from 1.6 ng/L to 3.2 ng/L. The average recoveries ranged from 86.2% to 105% and the RSDs from 3.8% to 9.4%. Conclusion The method is sensitive, fast and simple. It has the advantage of little organic solvent consumption, which is friendly to environment and suitable for the detection of seven PAHs in water.

Rapid chemome profiling of chemical components of Lonicerae Japonicae Flos using DI-MS/MS~(ALL) / 中国中药杂志

A new method of MS/MS~(ALL) was designed to sequentially record a MS~2 spectrum at each unit mass window through gas phase fractionation concept, so as to offer an opportunity for universal MS~2 spectral recording with direct infusion(DI). As a proof-of-concept, DI-MS/MS~(ALL) was applied for rapid chemome profiling of a famous herbal medicine named Lonicerae Japonicae Flos. After each MS~2 spectrum was correlated to its precursor ion, the structural annotation was conducted by applying well-defined mass cracking rules, matching the mass spectral data with literatures and referring to those accessible databases. As a result, a total of 54 components were identified from Lonicerae Japonicae Flos extract, including 21 phenolic acids, 13 flavonoids, 12 iridoids, 4 triterpenoids and 4 other compounds. Therefore, DI-MS/MS~(ALL) is a powerful tool for comprehensive, rapid qualitative analysis of chemical profiles of traditional Chinese medicine and other chemical components of complex systems.

[Cloning and expression analysis of CpPMM gene in Codonopsis pilosula].

GDP-mannose is an important precursor for the synthesis of Codonopsis pilosula polysaccharides and involved in the synthesis of sugar chains. Phosphomannomutase(PMM)catalyzes the conversion of mannose-6-phosphate(Man-6-P)to mannose-1-phosphate(Man-1-P)to synthesize GDP-mannose. In this study, specific primers were designed based on the PMM gene sequence information in transcriptome data, and the full length of the C. pilosula PMM gene was cloned and named CpPMM. The correlation between the CpPMM gene expression and C. pilosula polysaccharide synthesis was analyzed by a series of bioinformatics analysis, prokaryotic expression and qRT-PCR. The results show that the CpPMM gene contains a 741 bp open reading frame(ORF), encoding 246 amino acids, which is highly similar to the PMM of other species and highly homologous to the Helianthus annuus from the Asteraceae family. It was predicted to be a hydrophilic non-transmembrane protein without signal peptide, which was predicted to be located in the cytoplasm with multiple phosphorylation sites. Combined with predictive analysis of conserved domains, this protein belongs to the HAD(haloacid dehalogenase)superfamily; prokaryotic expression studies show that the size of the CpPMM fusion protein is about 29 kDa, which is consistent with the relative molecular mass predicted. The target protein is an inclusion body and is partially soluble. The qRT-PCR results showed that the CpPMM gene exerted spatiotemporal expression patterns, and the expression level in fruiting period was significantly higher than that in the other three periods such as the flowering period. Along with the growth period of C. pilosula, the polysaccharide content of C. pilosula showed a gradual increase trend, reaching the highest during the harvest time. And there are significant differences in the polysaccharide content of C. pilosula in each period. In this study, the CpPMM gene was cloned from the root of C. pilosula, at the same time, the prokaryotic expression system was constructed. In addition, its gene expression level is highly correlated with the polysaccharide content of C. pilosula. It lays the foundation for further studying the function of CpPMM gene and the analysis of biosynthetic pathways of polysaccharides in medicinal plants.

Online energy-resolved MS boosts the potential of LC-MS towards metabolite characterization of salidroside and tyrosol.

Although currently serving as the workhorse for metabolite characterization, one of the most challenging tasks for LC-MS is isomeric differentiation because isomers frequently yield identical quasi-molecular ions and fragmented ion species. Our previous studies have demonstrated that online energy-resolved MS (ER-MS) is an orthogonal technique for MS/MS experiments to facilitate isomeric identification. Herein, attempts were made for the in-depth characterization of the metabolic profiles of an effective natural product named salidroside (SA) in rats using LC coupled with three-dimensional mass spectrometry (LC-3D MS) that was configured by MS1, MS2 and online ER-MS as 1st, 2nd, and 3rd dimensions, respectively. Moreover, the metabolism characterization of its aglycone, namely, tyrosol (Try) was conducted in parallel to aid in proposing metabolic pathways. High-resolution MS1 and MS2 spectra were acquired by IT-TOF-MS, and subsequent data processing provided theoretical formula and sub-structures for each metabolite. Subsequently, online ER-MS was conducted for precursor > product ion transitions-of-interest to offer linkage information among the sub-structures via building breakdown graphs. As a result, ten (M1-10) and nine (M1, M2, and M5-11) metabolites were detected in SA- and Tyr-administrated biological samples, respectively, and their structures were qualitatively identified. Crucial metabolism occurred for either component. SA initially underwent hydrolysis to produce Tyr, and subsequently hydroxylation, oxidation, glucuronidation, and sulfation were observed as the primary metabolic pathways. To summarize, the metabolic fate of SA was understood in depth, and Tyr, as the hydrolytic product, was responsible for the occurrences of most metabolites (M1, M2, and M5-10). More importantly, identification confidences of the metabolites were significantly advanced by LC-3D MS, suggesting that it is eligible to serve as an integral part of the analyst's toolbox.

Psychiatric side effects induced by chloroquine and hydroxychloroquine: a systematic review of case reports and population studies

Chloroquine and hydroxychloroquine are commonly used drugs in the treatment of malaria as well as chronic diseases, such as rheumatoid arthritis, and systemic lupus erythematosus. Although various reports on possible psychiatric side effects of these drugs exist, the nature and extent of these effects remain poorly understood. Moreover, the relevance of these drugs in the treatment of early stages of COVID-19 necessitates a careful estimation of their side effects. Here, we provide a systematic review of the psychiatric side effects associated with chloroquine and hydroxychloroquine. We used PubMed, Scopus, and Web of Science platforms to identify relevant literature published between 1962 and 2020. Search terms included chloroquine, hydroxychloroquine, psychiatry, psychosis, depression, anxiety, bipolar disorder, delirium, and psychotic disorders. Only case reports and clinical trials were included. All studies included records of psychiatric side effects induced by either chloroquine or hydroxychloroquine or both. Both retrospective and prospective, randomized as well as non-randomized population studies were included. Overall, the psychiatric side effects are dose- and sex-independent. The most common psychiatric side effects reported are increased speech output/ excessive talking, increased psychomotor activity, irritable mood, auditory hallucinations, delusion of grandiosity, and suicide attempts, likely due to brain intoxicationbe of chloroquine or hydroxychloroquine. The symptoms can develop in a few hours to 11 weeks after drug intake and are normally reversed within a week after the drug withdrawal. We conclude that CQ and HCQ have the potential to induce psychiatric side effects. This study calls for further investigation of psychiatric symptoms induced by these drugs in the short and long term.

HPLC Fingerprint Analysis of Liposoluble and Water-soluble Fractions of Xiaojinwan / 中国实验方剂学杂志

Objective:High performance liquid chromatography (HPLC) fingerprints of liposoluble and water-soluble fractions of Xiaojinwan were established and the similarity of fingerprints was evaluated, so as to explore the quality consistency of Xiaojinwan. Method:Chromatographic separation was carried out on Welch Ultimate AQ-C18 column (4.6 mm×250 mm, 5 μm) with the mobile phase of 0.1% phosphoric acid solution (A)-acetonitrile (B) for gradient elution (liposoluble fraction of 0-5 min, 40%B; 5-10 min, 40%-50%B; 10-20 min, 50%-60%B; 20-30 min, 60%-65%B; 30-40 min, 65%-70%B; 40-50 min, 70%-80%B; 50-60 min, 80%-90%B; 60-65 min, 90%-95%B; 65-75 min, 95%-100%B; 75-80 min, 100%B; water-soluble fraction of 0-20 min, 2%-5%B; 20-30 min, 5%-10%B; 30-37 min, 10%-20%B; 37-45 min, 20%-30%B; 45-50 min, 30%-40%B; 50-58 min, 40%B), the flow rate was 1 mL·min-1, the column temperature was 30 ℃. The detection wavelengths of the liposoluble and water-soluble fractions were 202, 250 nm, and their injection volumes were 10, 20 μL, respectively. A total of 30 batches of Xiaojinwan from five manufacturers were detected by HPLC, the chromatographic peaks of each part were analyzed by principal component analysis (PCA) and identified. Result:A total of 55 chromatographic peaks were detected in the fingerprints, and the similarity of fingerprint of 30 batches of Xiaojinwan was quite different. The relative standard deviations (RSDs) of fingerprint similarity of liposoluble and water-soluble fractions of Xiaojinwan were 21.5% and 32.8%, respectively. There were significant differences in the quality of samples from different manufacturers and the same manufacturer, and the chemical consistency evaluation results were dominated by liposoluble fraction, and the main reason for the chemical difference of this preparation was the composition of Liquidambaris Resina. Conclusion:The quality consistency of Xiaojinwan is poor. The establishment of two-fraction fingerprint provides a new idea for the overall quality evaluation and control of Xiaojinwan, and can provide a reference for the quality consistency evaluation of traditional pills.

Quality evaluation of Xiaojin Pills based on antiplatelet aggregation biological potency assay / 中草药

Objective: To establish a biological potency assay for Xiaojin Pills against platelet aggregation in vitro, evaluate the quality consistency of Xiaojin Pills, and screen traditioanal Chinese medicines which play the role of promoting blood circulation in Xiaojin Pills. Methods: Xiaojin Pills and ten Chinese medicines [artificial musk, Momordica cochinchinensis, Aconitum kusnezoffii, Liquidambar formosana, Boswellia carterii, Commiphora myrrha, Faeces Trogopterori, Angelica sinensis, Pheretima aspergillum, Fragrant Ink] in its formula were extracted by ultrasound in 40% methanol. The antiplatelet aggregation rate of the extract was measured by platelet aggregation meter. The platelet-rich plasma (PRP) and platelet-poor plasma (PPP) were prepared from abdominal aorta of rats. The platelet aggregation was induced by adenosine diphosphate (ADP). With sodium ferulate as a standard reference material, the biological potency of antiplatelet aggregation of Xiaojin Pills was calculated by the simplified probit principle. Results: The results showed that the biological potency of Xiaojin Pills was between 0.598 and 1.338 U/mg among different manufacturers and batches. In Xiaojin Pills group, Pheretima, Faeces Trogopterori, and Momordicae Semen had stronger inhibitory effects on platelet aggregation with inhibition rates of 70.87%, 31.83% and 67.52%, respectively. Conclusion: The quality consistency of Xiaojin Pills from different manufacturers and batches is poor, and Pheretima, Faeces Trogopterori, and Momordicae Semen may be the key drugs for Xiaojin Pills to play the role of promoting blood circulation.