Research progress on stable gene transfection methods of Plasmodium falciparum / 中国热带医学

@#Abstract: Transfection of Plasmodium falciparum is helpful to study the function of its genes, such as drug resistance. However, transgenic manipulation has been very challenging, mainly due to the high A/T base sequence structure (A+T content of about 82%) and low transfection efficiency of the Plasmodium genome. Electroporation-based transfection of Plasmodium falciparum has been successfully applied in the study of certain genes, and electroporation by preloading is currently the preferred method for introducing foreign DNA into Plasmodium falciparum. The site-directed editing of Plasmodium genes mostly adopts the method of two-plasmid transfection. It is generally believed that successful transfection of Plasmodium requires a large amount of high-purity plasmid DNA and an accurate transfection system. In addition to the evaluation of the current commonly used electrotransfection methods, this paper also introduces a new transfection method, namely lyse-reseal erythrocytes for transfection (LyRET). This paper also review the role of factors such as plasmid DNA concentration, the use of transfection reagents, the setting of transfection parameters, the addition of fresh red blood cells, and the markers of successful transfection in improving the success rate and efficiency of Plasmodium transfection, in the hope of providing a reference for study in this field.

Epidemiological characteristics of pesticide poisoning in Chengdu City from 2012 to 2021 / 中华劳动卫生职业病杂志

Objective: To explore the present situation and epidemiological characteristics of pesticide poisoning in Chengdu City from 2012 to 2021, and to provide scientific evidence for further prevention and control. Methods: In January 2022, the pesticide poisoning report cards of Chengdu City from 2012 to 2021 were collected from the China Disease Control and Prevention Information System. The data of the report card was reorganized and the distribution characteristics of pesticide poisoning such as time, region, gender, age and pesticide types were analyzed. Results: 14326 cases of pesticide poisoning were reported in Chengdu City from 2012 to 2021, 651 deaths, and the fatality rate was 4.54%. The cases of productive pesticide poisoning and unproductive pesticide poisoning were 504 and 13822, respectively. The fatality rates of productive and unproductive pesticide poisoning were 1.39% and 4.66%, which were significant different (χ(2)=11.99, P=0.001). The highest reported cases of pesticide poisoning was in 2013 (1779) and the lowest in 2021 (1047). The number of reported cases showed a downward trend year by year (t=-12.30, P<0.001), and the fatality rates also showed a downward trend year by year (χ(2)(trend)=25.48, P<0.001). The fluctuation range of unproductive pesticide poisoning cases in each month of the year was small, and the productive pesticide poisoning mainly occurred from May to August. The regions with the largest number of reported poisoning cases were Pengzhou (1620), Jianyang (1393), Jintang (1266) and Qionglai (1158). The high incidence of poisoning was among 25-54 years old (50.21%, 7193/14326). The fatality rate in the age group 75-96 years old was the highest (8.98%, 95/1058), and the fatality rates increased gradually with age (χ(2)(trend)=186.03, P<0.001). The pesticides causing poisoning were mainly insecticide (43.86%, 6284/14326) and herbicides (35.75%, 5121/14326). Herbicides paraquat had the highest fatality rate (9.54%, 286/2998) . Conclusion: Pesticide poisoning in Chengdu City is mainly unproductive poisoning. Health education should be carried out for key areas and people, and the control of highly toxic pesticides such as insecticide and herbicides should be strengthened.

HIV self-testing and related factors in men who have sex with men in Shijiazhuang / 中华流行病学杂志

Objective: To understand HIV self-testing and related factors in men who have sex with men (MSM) in Shijiazhuang. Methods: From August to September 2020, convenient sampling was used to recruit MSM in Shijiazhuang. Online questionnaires were used to collect information about their demographic characteristics, sexual behaviors and HIV self-testing. logistic regression model was used to analyze the related factors associated with HIV self-testing. Results: In the 304 MSM respondents, 52.3% (159/304) had HIV self-testing in the past 6 months, and 95.0% (151/159) used fingertip blood HIV detection reagent. Self-purchase was the main way to obtain HIV testing reagents (45.9%, 73/159), followed by supply from MSM social organization (44.7%, 71/159). The reasons for having HIV self-testing were non-specific testing time (67.9%, 108/159) and privacy protection (62.9%,100/159), the reasons for having no HIV self-testing included inability of using (32.4%, 47/145), being unaware of HIV self-testing reagent (24.1%, 35/145), and worry about inaccurate self-testing results (19.3%, 28/145). Multivariate logistic regression analysis showed that being 18-29 years old (aOR=2.68, 95%CI: 1.20-5.94), obtaining free HIV self-testing kits in recent 6 months (aOR=8.61, 95%CI: 4.09-18.11) and making friends through Internet and social software (aOR=2.68, 95%CI: 1.48-4.88) were positive factors for having HIV self-testing. Conclusion: HIV self-testing is a more flexible and convenient way to detect HIV in MSM, and the promotion of HIV self-testing in MSM should be strengthened to further increase the HIV detection rate in this population.

HbA1c comparison and diagnostic efficacy analysis of multi center different glycosylated hemoglobin detection systems / 中华预防医学杂志

Objective: Compare and analyze the results of the domestic Lanyi AH600 glycated hemoglobin analyzer and other different detection systems to understand the comparability of the detection results of different detectors, and establish the best cut point of Lanyi AH600 determination of haemoglobin A1c (HbA1c) in the diagnosis of diabetes. Methods: Multi center cohort study was adopted. The clinical laboratory departments of 18 medical institutions independently collected test samples from their respective hospitals from March to April 2022, and independently completed comparative analysis of the evaluated instrument (Lanyi AH600) and the reference instrument HbA1c. The reference instruments include four different brands of glycosylated hemoglobin meters, including Arkray, Bio-Rad, DOSOH, and Huizhong. Scatter plot was used to calculate the correlation between the results of different detection systems, and the regression equation was calculated. The consistency analysis between the results of different detection systems was evaluated by Bland Altman method. Consistency judgment principles: (1) When the 95% limits of agreement (95% LoA) of the measurement difference was within 0.4% HbA1c and the measurement score was≥80 points, the comparison consistency was good; (2) When the measurement difference of 95% LoA exceeded 0.4% HbA1c, and the measurement score was≥80 points, the comparison consistency was relatively good; (3) The measurement score was less than 80 points, the comparison consistency was poor. The difference between the results of different detection systems was tested by paired sample T test or Wilcoxon paired sign rank sum test; The best cut-off point of diabetes was analyzed by receiver operating characteristic curve (ROC). Results: The correlation coefficient R2 of results between Lanyi AH600 and the reference instrument in 16 hospitals is≥0.99; The Bland Altman consistency analysis showed that the difference of 95% LoA in Nanjing Maternity and Child Health Care Hospital in Jiangsu Province (reference instrument: Arkray HA8180) was -0.486%-0.325%, and the measurement score was 94.6 points (473/500); The difference of 95% LoA in the Tibetan Traditional Medical Hospital of TAR (reference instrument: Bio-Rad Variant II) was -0.727%-0.612%, and the measurement score was 89.8 points; The difference of 95% LoA in the People's Hospital of Chongqing Liang Jiang New Area (reference instrument: Huizhong MQ-2000PT) was -0.231%-0.461%, and the measurement score was 96.6 points; The difference of 95% LoA in the Taihe Hospital of traditional Chinese Medicine in Anhui Province (reference instrument: Huizhong MQ-2000PT) was -0.469%-0.479%, and the measurement score was 91.9 points. The other 14 hospitals, Lanyi AH600, were compared with 4 reference instrument brands, the difference of 95% LoA was less than 0.4% HbA1c, and the scores were all greater than 95 points. The results of paired sample T test or Wilcoxon paired sign rank sum test showed that there was no statistically significant difference between Lanyi AH600 and the reference instrument Arkray HA8180 (Z=1.665,P=0.096), with no statistical difference. The mean difference between the measured values of the two instruments was 0.004%. The comparison data of Lanyi AH600 and the reference instrument of all other institutions had significant differences (all P<0.001), however, it was necessary to consider whether it was within the clinical acceptable range in combination with the results of the Bland-Altman consistency analysis. The ROC curve of HbA1c detected by Lanyi AH600 in 985 patients with diabetes and 3 423 patients with non-diabetes was analyzed, the area under curve (AUC) was 0.877, the standard error was 0.007, and the 95% confidence interval 95%CI was (0.864, 0.891), which was statistically significant (P<0.001). The maximum value of Youden index was 0.634, and the corresponding HbA1c cut point was 6.235%. The sensitivity and specificity of diabetes diagnosis were 76.2% and 87.2%, respectively. Conclusion: Among the hospitals and instruments currently included in this study, among these four hospitals included Nanjing Maternity and Child Health Care Hospital in Jiangsu Province (reference instrument: Arkray HA8180), Tibetan Traditional Medical Hospital of TAR (reference instrument: Bio-Rad Variant Ⅱ), the People's Hospital of Chongqing Liang Jiang New Area (reference instrument: Huizhong MQ-2000PT), and the Taihe Hospital of traditional Chinese Medicine in Anhui Province (reference instrument: Huizhong MQ-2000PT), the comparison between Lanyi AH600 and the reference instruments showed relatively good consistency, while the other 14 hospitals involved four different brands of reference instruments: Arkray, Bio-Rad, DOSOH, and Huizhong, Lanyi AH600 had good consistency with its comparison. The best cut point of the domestic Lanyi AH600 for detecting HbA1c in the diagnosis of diabetes is 6.235%.

Analysis of pathogenic bacteria distribution and drug resistance characteristics of bloodstream infection in patients with neutrophilic deficiency after chemotherapy in acute leukemia / 中国热带医学

@#Abstract: Objective　To investigate the distribution and drug resistance characteristics of pathogenic bacteria in patients with neutropenic acute leukemia (AL) and bloodstream infections (BSI). Methods　The clinical data of 258 neutropenic acute leukemia patients with bloodstream infections, who admitted to Shengjing Hospital of China Medical University from January 2016 to December 2021, were collected and analyzed for pathogenic bacteria and drug resistance. Results A total of 268 strains of pathogenic bacteria were isolated from 258 patients, including 180 strains of gram-negative bacteria (67.16%), 61 strains of gram-positive bacteria (22.76%), and 27 strains of fungi (10.07%). Gram-negative bacteria were mainly Klebsiella pneumoniae (53/268, 19.78%), Escherichia coli (49/268, 18.28%) and Pseudomonas aeruginosa (41/268, 15.30%). Gram-positive bacteria were mainly coagulase negative Staphylococcus (31/268, 11.57%) and Staphylococcus aureus(17/268, 6.34%). The main fungi were Candida tropicalis (25/268, 9.33%). Escherichia coli (33/268, 12.31%) was the most common pathogen isolated from acute myeloid leukemia (AML), followed by Pseudomonas aeruginosa (25/268, 9.33%), coagulase-negative Staphylococcus (18/268, 6.72%) and Candida tropicalis (18/268, 6.72%). Klebsiella pneumoniae (35/268, 13.06%) was the most common pathogen isolated from acute lymphoblastic leukemia (ALL),followed by Pseudomonas aeruginosa (15/268, 5.60%) and Escherichia coli (14/268, 5.22%). The resistance of Gram-negative bacteria to piperacillin/tazobactam, cefoperazone/sulbactam, imipenem, meropenem, ertapenem, amikacin, cefoxitin, amoxicillin/clavulanic acid was low. Gram-positive bacteria were sensitive to linezolid and vancomycin. Candida was sensitive to flucytosine, amphotericin B and itraconazole. Conclusions　In patients with granulosa after AL chemotherapy combined with BSI, the pathogenic bacteria isolated from AML are diverse, and the pathogenic bacteria isolated from ALL are mainly gram-negative bacteria. Pathogenic bacteria have different degrees of drug resistance to commonly used antibacterial drugs, so it is important to strengthen the monitoring of the distribution of pathogenic bacteria and the change of drug resistance and rational use of antibacterial drugs to minimize the death of patients.

Optimization of β-globin Stable Expression Using the Third Generation Lentiviral Vector for β-thalassemia Therapy / 中国实验血液学杂志

OBJECTIVE@#To provide a research basis for a safe and effective cell therapy for β-thalassemia through optimization of HS4 region of the third generation lentiviral vector for stable expression of β-globin.@*METHODS@#The human β-globin HS4 region in the third generation lentiviral expression vector was optimized to construct the lenti-HBB, and the transcription and translation of β-globin gene were analyzed by RT-PCR and Western blot after the transduction of lenti-HBB in MEL cell line. Furthermore, the erythroid differentiation of CD34+ cells which were transduced lentiviral virus carrying human β-globin from normal human umbilical cord blood cells and peripheral blood cells of patients with β-thalassemia major were confirmed by colony formation assay, cell smear assay and flow cytometry. The safety and effectiveness of the optimized lenti-HBB were verified by NSG mouse in vivo test.@*RESULTS@#The human β-globin was expressed stably in the MEL cells, and CD34+ cells from health umbilical cord blood as well as PBMC from patient with β-thalassemia major transduced with lenti-HBB could be differentiated to mature red blood cells. The β-globin expression and differentiation in CD34+ cells were demonstrated successfully in the NSG mouse for about 35 months after post-transplant.@*CONCLUSION@#Stable β-globin expression through the optimization of HS4 from CD34+ in the third generation lentiviral vector is safe and effective for patients with severe β-thalassemia and other β-globin abnormal diseases.

Molecular Detection of Tick-borne Pathogens in Ticks Collected from Hainan Island, China / 生物医学与环境科学（英文）

Pathogens like bacteria and protozoa, which affect human and animal health worldwide, can be transmitted by vectors like ticks. To investigate the epidemiology and genetic diversity of bacteria and protozoans carried by ticks in Chengmai county of Hainan province, China, 285 adult hard ticks belonging to two species [

Effect of peritoneum reconstruction on postoperative complications after laparoscopic low anterior resection for rectal cancer / 中华胃肠外科杂志

Objective: To determine the effect of peritoneum reconstruction on postoperative complications after laparoscopic low anterior resection (LAR) for rectal cancer. Methods: Retrospective cohort study and propensity score matching were conducted. Case inclusion criteria: (1) pathologically confirmed rectal adenocarcinoma; (2) 18 to 80 years; (3) patients with middle to low rectal cancer undergoing laparoscopic LAR; (4) patients staging cT1-4aN0-2M0 or ycT1-4aN0-2M0 after neoadjuvant therapy; (5) the distance of 4-10 cm from tumor low margin to anal verge. Exclusion criteria: (1) abdominal surgery history (except appendicitis, cholecystitis, ectopic pregnancy); (2) anastomosis above the peritoneal reflection; (3) tumor distant metastasis or clinical staging of T4b during surgery; (4) conversion to open surgery; (5) severe incapacitating disease (American Society of Anesthesiologists classification IV or V, ASA). A total of 666 patients with middle to low rectal cancer undergoing laparoscopic LAR in The First Affiliated Hospital of Army Medical University from January 2017 to June 2020 were enrolled. There were 473 males and 193 females with the median age of 59 (18-80) years. Laparoscopic LAR with peritoneum reconstruction was performed in 188 cases (PR group), and laparoscopic LAR without peritoneum reconstruction was performed in 478 cases (NPR group). After 1:1 propensity score matching according to 1:1 based on age, gender, body mass index, TNM staging, ASA classification, intraoperative blood loss, distance from tumor low margin to anal edge, 153 cases were included in each group. Postoperative complications were classified according to Clavien-Dindo classification. Anastomotic leakage was defined and graded according to the International Study Group of Rectal Cancer (ISGRC) criteria. Results: After propensity score matching, there were no significant differences in baseline demographic characteristics between the 2 groups (all P>0.05), indicating that these two groups were comparable. (1) Operative conditions: All the patients in both groups completed operation successfully. Compared with the NPR group, the PR group had longer operation time [(181.3±60.3) minutes vs. (168.9±51.5) minutes, t=2.185, P=0.029], shorter postoperative median hospital stay [8 (7, 10) days vs. 9 (7, 11) days, Z=-2.282, P=0.022], and the differences were statistically significant (P<0.05). (2) Postoperative complications: The overall morbidity of postoperative complication in PR group and NPR group was 20.3% (31/153) and 24.2% (37/153) respectively, and the incidence of anastomotic leakage was 9.8% (15/153) and 11.1%(17/153) respectively, whose differences were not statistically significant (both P>0.05). Compared with NPR group, PR group had lower morbidity of grade III to IV complications [3.9% (6/153) vs. 11.1% (17/153), χ(2)=5.688, P=0.017] and lower secondary operation rate [1.3% (2/153) vs. 5.9% (9/153), χ(2)=4.621, P=0.032], the differences were statistically significant (both P<0.05). Though PR group had lower incidence of grade C anastomoic leakage [1.3% (2/153) vs. 3.9% (6/153), χ(2)=2.054, P=0.152], but the differences were not statistically significant. (3) Postoperative inflammation: The difference of the procalcitonin level of both PR and NPR groups at postoperative 1-d, 3-d, and 5-d was statistically significant (F=5.222, P=0.010) in time-dependent manner, while the difference was not significant in the interaction effect (P>0.05). No statistically significant differences in the C-reactive protein level between two groups at postoperative 1-d, 3-d, and 5-d were found (all P>0.05). Conclusion: Peritoneum reconstruction in laparoscopic LAR can decrease the morbidity of postoperative complication of grade III to IV and the reoperation rate, and plays an important role in controlling the inflammatory reaction, which has great clinical value.

Phase II Study of Preoperative IntraArterial Epirubicin, Etoposide, and Oxaliplatin Combined with Oral S-1 Chemotherapy for the Treatment of Borrmann Type 4 Gastric Cancer

Purpose@#A phase II study was conducted to evaluate the safety and efficacy of preoperative, intra-arterial perfusion of epirubicin, etoposide, and oxaliplatin combined with oral chemotherapy S-1 (SEEOX) for the treatment of type 4 gastric cancer. @*Materials and Methods@#A single-center, single-arm phase II trial was conducted on 36 patients with histologically proven type 4 gastric cancer without distant peritoneal or organ metastasis. Patients received 3, 21-day courses of SEEOX preoperative chemotherapy. The primary endpoint was overall survival (OS) and the secondary outcomes assessed were chemotherapeutic response, radical resection rate, pathological regression, toxicities, postoperative morbidity, and mortality. @*Results@#All patients were at an advanced stage of cancer (stage III or IV) and completed the entire course of treatment. Based on changes in tumor volume and peritoneal metastasis, the objective response rate was 55.6% (20/36; 95% confidence interval [CI], 38.5%–72.6%) and the disease control rate was 69.4% (25/36; 95% CI, 53.6%–85.3%). The radical resection rate was 75% (27/36; 95% CI, 60.1%–89.9%) and the proportion of R0 resections was 66.7% (21/36; 95% CI, 50.5%–82.8%). The pathological response rate was 33.3%, of which 13.9% showed complete pathological regression. The median survival was 27.1 months (95% CI, 22.24–31.97 months), and the 2-year OS was 48.5% (95% CI, 30.86%–66.1%). @*Conclusions@#Preoperative SEEOX is a safe and effective treatment for type 4 gastric cancer. Based on these preliminary data, a phase III study will be conducted to confirm the superiority of this regimen over standard treatment.Trial Registration: ClinicalTrials.gov Identifier: NCT02949258

Analysis of Saliva Microbiome in Patients with Esophageal Squamous Cell Carcinoma / 中山大学学报(医学科学版)

@#【Objective】To analyze the difference of salivary microbiome between patients with esophageal squamous cell carcinoma（ESCC group）and healthy controls（HC group），and to screen out the target bacteria in saliva of patients with esophageal squamous cell carcinoma. 【Methods】 Salivary samples were collected from 50 patients with esophageal squamous cell carcinoma and 40 healthy controls. Total DNA was extracted，16S rDNA V4 region was amplified by PCR，high-throughput sequencing was performed，and sequencing data was subjected to OUT clustering，species annotation and diversity analysis.【Results】Diversity analysis showed difference in the diversity of the salivary microbiome between the ESCC group and the HC group，but P > 0.05. Species analysis showed that the salivary microbiome in the ESCC group was dominated by Neisseria，Fusobacterium，Haemophilus，Prevotella and Porphyromonas，while Neisseria，Haemophilus， Prevotella，Fusobacterium and Veillonella were the top microbiota in the HC group. LEfse analysis showed that the expression of Porphyromonas in the ESCC group was elevated and the result had statistical significance.【Conclusion】Saliva in the patients with esophageal squamous cell carcinoma has characteristic microbiome composition. Compared with the squamous cell carcinoma is significantly increased，and the result is statistically significant(P < 0.05）.

Phase II Study of Preoperative IntraArterial Epirubicin, Etoposide, and Oxaliplatin Combined with Oral S-1 Chemotherapy for the Treatment of Borrmann Type 4 Gastric Cancer

Purpose@#A phase II study was conducted to evaluate the safety and efficacy of preoperative, intra-arterial perfusion of epirubicin, etoposide, and oxaliplatin combined with oral chemotherapy S-1 (SEEOX) for the treatment of type 4 gastric cancer. @*Materials and Methods@#A single-center, single-arm phase II trial was conducted on 36 patients with histologically proven type 4 gastric cancer without distant peritoneal or organ metastasis. Patients received 3, 21-day courses of SEEOX preoperative chemotherapy. The primary endpoint was overall survival (OS) and the secondary outcomes assessed were chemotherapeutic response, radical resection rate, pathological regression, toxicities, postoperative morbidity, and mortality. @*Results@#All patients were at an advanced stage of cancer (stage III or IV) and completed the entire course of treatment. Based on changes in tumor volume and peritoneal metastasis, the objective response rate was 55.6% (20/36; 95% confidence interval [CI], 38.5%–72.6%) and the disease control rate was 69.4% (25/36; 95% CI, 53.6%–85.3%). The radical resection rate was 75% (27/36; 95% CI, 60.1%–89.9%) and the proportion of R0 resections was 66.7% (21/36; 95% CI, 50.5%–82.8%). The pathological response rate was 33.3%, of which 13.9% showed complete pathological regression. The median survival was 27.1 months (95% CI, 22.24–31.97 months), and the 2-year OS was 48.5% (95% CI, 30.86%–66.1%). @*Conclusions@#Preoperative SEEOX is a safe and effective treatment for type 4 gastric cancer. Based on these preliminary data, a phase III study will be conducted to confirm the superiority of this regimen over standard treatment.Trial Registration: ClinicalTrials.gov Identifier: NCT02949258

Hypofibrinogenemia Caused by Hemocoagulase Injection: A Retrospective Study on Clinical Laboratory Findings / 中国医学科学杂志(英文版)

Objective Hemocoagulase injection based on the venom of is widely used in the treatment of hemorrhagic disorders. This study aimed to characterize the clinical laboratory findings of hemocoagulase-induced hypofibrinogenemia as the associated adverse reaction of hemocoagulase injection.Methods We retrospectively enrolled 27 in-patients who were treated with hemocoagulase injection for hemoptysis and developed hypofibrinogenemia during the period of January 1, 2015 to March 31, 2018. Clinical data were collected and investigated, including clinical manifestations, hemostatic and fibrinolytic parameters, dosage of hemocoagulase, the medication time, and the cryoprecipitate blood product infusion. Differences in fibrinogen, D-dimer, and fibrin/fibrinogen degradation products (FDP) before, during, and after the application of hemocoagulase injection were analyzed statistically.Results Plasma fibrinogen level during medication of hemocoagulase injection decreased significantly compared to that before the treatment (=1.80, <0.001), with the average decrease of 2.28 g/L (0.63-3.9 g/L). After withdrawal, fibrinogen level increased significantly compared to that during the medication (=-1.20, <0.001), but was still lower than that before the medication (=0.59, =0.03). The D-dimer level and the FDP level after withdrawal decreased significantly compared to the levels during the medication (=0.83, =0.002; Wilcoxon-test, =-4.54, <0.001). Spearman's correlation analyses did not find either fibrinogen change during-before the administration or FDP change after-during the administration was associated with the dosage of hemocoagulase (=-0.17, =0.40; =-0.28, =0.15; respectively) and the time of recovery from hypofibrinogenemia (=-0.45, =0.05; =0.13, =0.61; respectively).Conclusion Monitoring both clotting and fibrinolysis parameters is essential in the management of hemoptysis patients treated with hemocoagulase injection. Clinicians should be aware of hypofibrinogenemia and consider discontinuation of the administration of hemocoagulase whenever necessary.

The inhibition effects of allicin on TGF-β1 induced epithelial⁃mesenchymal transition of human cholangiocarcinoma cell and its mechanism / 医学研究生学报

Objective　The metastasis mechanism of cholangiocarcinoma is complex, which may be related to epithelial-mesenchymal transition(EMT). This study focused on investigating the inhibition effects of allicin on TGF-β1 induced epithelium mesenchymal transition of human cholangiocarcinoma cells and its related mechanism, and providing theoretical basis for the application of allicin in the treatment of cholangiocarcinoma.　Methods　MTT assay were used to detect the inhibition effects of different concentrations of allicin on the human cholangiocarcinoma RBE cell proliferation, and the drug concentration of allicin was determined by IC50 of 24 h. The RBE cells were cultured and divided into control group, allicin group(130.7μmol/L), TGF-β1 group(10ng/mL) and allicin+ TGF-β1 group(130.7μmol/L+10ng/mL). Wound scratch and transwell invasion assay were performed to detect the migration and invasion ability of RBE cells after 24 hours. Western blots were applied to detect expression of EMT-related proteins (E-Cadherin, N-Cadherin, Vimentin, Snail) and NF-κB signaling pathways.　Results　The migration rates in allicin group and allicin+ TGF-β1 group were both decreased compared with that in the control group ( 9.25% ± 0.36% vs 28.19 %±0.66%, P<0.05) and TGF⁃β1 group(13.91%±0.75% vs 49.22%±0.27%, P<0.05). The invasion rates in allicin group and allicin+ TGF-β1 group were also decreased compared with that in the control group (6.59%±0.06% vs 33.48%±0.04%, P<0.05) and TGF⁃β1 group(9.4%± 0.05% vs 40.21%±0.12%, P<0.05). Compared with the control group, E-Cadherin expression was significantly increased, and N-Cadherin, Vimentin, Snail, NF-κB and p-NF-κB expression were significantly decreased in the allicin group (P<0.05). Compared with TGF-β1 group, E-Cadherin expression was significantly up-regulated, and N-Cadherin, Vimentin, Snail, NF-κB and p-NF-κB expression were significantly down-regulated in the allicin+ TGF-β1 group (P<0.05)．　Conclusion　These results indicate that allicin can inhibit the EMT induced by TGF-β1 on the human cholangiocarcinoma cell by blocking NF-κB signaling pathway, which may have potential value to be the drug candidate for the treatment of human cholangiocarcinoma in future.

Essence of early gastric cancer and confusion in clinical practic / 中国实用外科杂志

Staging is a subjective assessment of objective progress of gastric cancer by doctors.However, the progress of gastric cancer is the result of tumor breaking through the limitation of immunity according to its inherent characteristics.The subjective evaluation methods are difficult to fully reflect the progress of gastric cancer. From the beginning of the concept of early gastric cancer to the wide application of endoscopic submucosal dissection(ESD) in clinic, the biggest puzzle is how to exclude lymph node metastasis. So far, it has not been satisfactorily solved. Therefore, in all previous Japanese Gastric Cancer Treatment Guidelines, the indications of ESD must be accompanied by the evaluation and treatment after endoscopic resection for possible residual cancer. Comprehending the essence of early gastric cancer and the related factors affecting its progress will help us to improve the quality of treatment for early gastric cancer and benefit patients.

Limited intervention measures for childbirth / 中国实用妇科与产科杂志

In the clinical work of our country,more and more attention is paid to the people-centered concept. It is more focuses on avoiding excessive intervention and strengthening management of labor course. Interventions that should be limited during childbirth for low-risk spontaneous labour.The limited interventions are including the application of the new parturition process criteria,the use of early amniotomy with early oxytocin augmentation for prevention of delay in labour is not recommended,routine cardiotocography is not recommended,encouraging the adoption of a birth position of the individual woman's choicey,routine episiotomy is not recommend,et al.Itis necessary to manage women and their fetus depending on patients' s situation,and to ensure the safety of mother and child.As well as to reduce excessive intervention,thereby reducing caesarean section rate,ensure that the mother has a good delivery experience,and improve the outcomes of mother and child.

Brief analysis on point of medicine and clinical application / 中国针灸

point of medicine is a set of point distributed as circle form, it is a common point in medicine. In this paper, point of medicine was analyzed from the following five aspects, i.e. basic theory, location, affiliation to heaven, earth and human being, main usage and principles of point combinations. medicine believes that the point is located in the middle of human body, where the essence of heaven and earth converges. point is closely connected with dragon and fire systems as well as organs in medicine, it acts on promoting circulation, adjusting and blood, and treating various diseases. It is highly valuable in clinical application.

An analysis of GNAS and THRA gene mutations in children with congenital hypothyroidism / 中国当代儿科杂志

OBJECTIVE@#To preliminarily investigate the relationship between stimulatory G protein α subunit (GNAS) and thyroid hormone receptor α (THRA) gene mutations and clinical phenotypes in children with congenital hypothyroidism (CH).@*METHODS@#A total of 70 children with CH diagnosed by neonatal screening were enrolled. Their peripheral blood samples were collected to extract genomic DNA. GNAS and THRA genes were screened for mutations using next-generation sequencing. Bioinformatics software was used to analyze the pathogenicity of gene mutations.@*RESULTS@#Of the 70 children with CH, nine missense mutations (three known mutations and six novel mutations) in the GNAS gene were detected in three patients (4%), and one gene polymorphism, c.508A>G(p.I170V), in the THRA gene was detected in four patients. The analysis results of bioinformatics software and ACMG/AMP guidelines showed that the two GNAS gene mutations [c.301C>T(p.R101C) and c.334G>A(p.E112K)] were more likely to be pathogenic. Three children with GNAS gene mutations showed different degrees of hypothyroidism.@*CONCLUSIONS@#GNAS gene mutations are related to the development of CH, and children with CH have different clinical manifestations. THRA gene mutations may not be associated with CH.

Features and therapeutic potential of T-cell receptors in high-grade glioma / 中华医学杂志(英文版)

BACKGROUND@#Previous studies have shown that endogenous T cells play an important role in the prolonged survival time of high-grade glioma (HGG) patients. Our objectives were to investigate the features of T-cell receptor (TCR) repertoires in HGG patients and to elucidate any potential therapeutic value.@*METHODS@#During November 2011 and December 2018, tumor tissues and blood samples of 35 patients with HGG who underwent surgery at Beijing Tiantan Hospital or Beijing Shijitan Hospital were selected after surgery. After isolating DNA from samples, multiple rounds of PCR were performed to establish a DNA immune repertoire (IR). Then, the sequences and frequencies of the complementarity-determining 3 (CDR3) region in TCR beta chain (TRB) were identified by high-throughput sequencing and IR analysis. A survival follow-up was conducted monthly thereafter until December 2018. Finally, the t test and Mann-Whitney test were used to compare statistical differences between two sets of data.@*RESULTS@#The Shannon diversity index (SHDI) of TRB sequences of HGG patients was significantly lower than that of healthy individuals (7.34 vs. 8.45, P = 0.001). The SHDI of TRB sequences of glioblastoma (GBM) patients with more than 16 months survival time was much higher than that of GBM patients with shorter survival times in both tumor tissues (3.48 ± 0.31 vs. 6.21 ± 0.33, t = -5.49, P = 0.002) and blood cells (6.02 ± 0.66 vs. 7.44 ± 0.32, t = -2.20, P = 0.036). In addition, patients achieved a distinctly higher proportion compared to that of healthy individuals in the proportion of TRBV9 and TRBV5 functional regions (9.83% vs. 6.83%, P = 0.001). Surgical tissue from patients who survived more than 16 months yielded a much higher proportion of TRBV4 and TRBV9 regions (7.14% vs. 3.28%, t = 3.18, P = 0.019). In surgical tissues from two GBM patients who survived for longer than 46 months, we found a potentially therapeutic TCR sequence.@*CONCLUSIONS@#HGG patients have less species diversity of TCR repertoires compared with that of healthy individuals. TRBV9 regions in TCRs may be protective factors for long-term survival of GBM patients.

Research Progress on Prevention and Control Measures of Aflatoxin in Chinese Medicinal Materials / 中国实验方剂学杂志

At present,traditional Chinese medicine(TCM) has attracted more and more attention from the international community.The demand for TCM is increasing in the world.The hidden dangers of potential quality and safety of TCM are also becoming increasingly prominent.Aflatoxin contamination has become one of the important factors affecting the safety of Chinese herbal medicines,and it will fundamentally affect human health and life safety.A variety of methods are used to reduce aflatoxins,however,there are few suitable methods that can be widely used in the cost-effective and large-scale promotion of Chinese herbal medicines.Therefore,it is of great significance to continue to study measures to solve the pollution problems of Aspergillus flavus and its toxins.This article summarizes the hazards and contamination status of aflatoxin,the prevention and control of the growth of A. flavus, and the measures for reducing aflatoxin,and looks ahead to the future prevention and control of A. flavus and its toxins,aiming at providing ideas for the pollution problem of A. flavus and its toxin,to ensure the quality of Chinese herbal medicines,so as to ensure clinical safety medication.

Reflections on Relationship Between Pathogenesis of Endometriosis and Intestinal Flora / 中国实验方剂学杂志

Endometriosis is a multiple disease that afflicts the health of women at childbearing age,and its incidence rate has been increasing year by year,furthermore,there has been a trend to be younger.At present,the pathogenesis of endometriosis has been not expounded completely,its cure rate is not high with high recurrence rate.In recent years,studies have shown that the human is a commensal body composed of a large number of microorganisms,and especially the microorganisms in the intestinal are closely related to the health of the body.Based on the previous studies on endometriosis,this paper proposes that its pathogenesis may be related to intestinal microbiological disorder,and aims to provide new ideas for the treatment of endometriosis.