Targeting microRNA-125b inhibited the metastasis of Alisertib resistance cells through mediating p53 pathway / 中华肿瘤杂志

Objective: To clarify the mechanisms involvement in Alisertib-resistant colorectal cells and explore a potential target to overcome Alisertib-resistance. Methods: Drug-resistant colon cancer cell line (named as HCT-8-7T cells) was established and transplanted into immunodeficient mice. The metastasis in vivo were observed. Proliferation and migration of HCT-8-7T cells and their parental cells were assessed by colony formation and Transwell assay, respectively. Glycolytic capacity and glutamine metabolism of cells were analyzed by metabolism assays. The protein and mRNA levels of critical factors which are involved in mediating glycolysis and epithelial-mesenchymal transition (EMT) were examined by western blot and reverse transcription-quantitative real-time polymerase chain reaction(RT-qPCR), respectively. Results: In comparison with the mice transplanted with HCT-8 cells, which were survival with limited metastatic tumor cells in organs, aggressive metastases were observed in liver, lung, kidney and ovary of HCT-8-7T transplanted mice (P<0.05). The levels of ATP [(0.10±0.01) mmol/L], glycolysis [(81.77±8.21) mpH/min] and the capacity of glycolysis [(55.50±3.48) mpH/min] in HCT-8-7T cells were higher than those of HCT-8 cells [(0.04±0.01) mmol/L, (27.77±2.55) mpH/min and(14.00±1.19) mpH/min, respectively, P<0.05]. Meanwhile, the levels of p53 protein and mRNA in HCT-8-7T cells were potently decreased as compared to that in HCT-8 cells (P<0.05). However, the level of miRNA-125b (2.21±0.12) in HCT-8-7T cells was significantly elevated as compared to that in HCT-8 cells (1.00±0.00, P<0.001). In HCT-8-7T cells, forced-expression of p53 reduced the colon number (162.00±24.00) and the migration [(18.53±5.67)%] as compared with those in cells transfected with control vector [274.70±40.50 and (100.00±29.06)%, P<0.05, respectively]. Similarly, miR-125b mimic decreased the glycolysis [(25.28±9.51) mpH/min] in HCT-8-7T cells as compared with that [(54.38±12.70)mpH/min, P=0.003] in HCT-8-7T cells transfected with control. Meanwhile, in comparison with control transfected HCT-8-7T cells, miR-125b mimic also significantly led to an increase in the levels of p53 and β-catenin, in parallel with a decrease in the levels of PFK1 and HK1 in HCT-8-7T cells (P<0.05). Conclusions: Silencing of p53 by miR-125b could be one of the mechanisms that contributes to Alisertib resistance. Targeting miR-125b could be a strategy to overcome Alisertib resistance.

Integration of history of medicine in the education and reform exploration of standardized residency training in endocrinology department — taking hundred years of history of insulin development as an example / 中华医学教育探索杂志

Objective:To study the methods of hormone-related teaching in standardized residency training of endocrinology.Methods:From February 2022 to June 2022, 25 residents in the Department of Endocrinology, Peking University Third Hospital were given a standardized residency training. The teaching mode combined with a hundred years of insulin development was adopted to practice insulin teaching, and questionnaires were evaluated before and after the teaching. Chi-square test was performed by SPSS 20.0.Results:A total of 31 physicians participated in the training, and 25 residents who were included in the first-stage standardized training were analyzed. Before the training, only 3 people (12%) knew the development history of insulin, and 10 people (40%) answered all the questions about clinical use of insulin correctly. After the training, 23 residents (92%) said they knew or were familiar with the history of insulin development, and 20 residents (80%) answered the questions about the clinical use of insulin correctly, all of which were higher than those before the training, and the difference was statistically significant.Conclusion:The integration of medical history in standardized residency training in endocrinology department can effectively improve the teaching effect, and enhance the humanity quality of young doctors, which can become an effective vehicle in standardized residency training.

Advance in hyperandrogenism and atherosclerotic disease in women / 中华内分泌代谢杂志

Hyperandrogenism is a common endocrine pathological state in women, and polycystic ovary syndrome is the main cause. Studies have shown that in addition to affecting reproductive function, hyperandrogenism in women can also interfere with vascular endothelial function, and directly or indirectly increases the risk of atherosclerotic disease by affecting risk factors such as blood pressure, lipids, and glucose. This article reviews the impact of hyperandrogenism on the cardiovascular system of women, aiming at a deeper understanding of the role of androgens in women′s health.

Effect of amantadine dimer adjuvant on humoral immune response induced by SARS-CoV-2 protein vaccine in mice / 中国生物制品学杂志

Effect of amantadine dimer adjuvant on humoral immune response induced by SARS-CoV-2 protein vaccine in mice@#Objective To investigate the effect of amantadine dimer adjuvant on humoral immune response induced by SARS-CoV-2 crown protein vaccine in mice.Methods The amantadine dimer was synthesized by substitution reaction ligation,hydrolytic acidification reaction ligation and amide condensation reaction ligation,with which as adjuvant,female BALB/c mice were immunized with the receptor-binding domain(RBD).The mice were randomly divided into five groups,six for each as follows:R6A+RBD group[21 μg(0.033 μmol)amantadine dimer+10 μg RBD],Ada+RBD group[10 μg(0.066 μmol)amantadine+10 μg RBD],Alu+RBD group(35 μg aluminum adjuvant+10 μg RBD),RBD group(10 μg RBD)and Blank group(0.9% normal saline),which were immunized i.m.on day 0,14 and 28 respectively.Serum samples were collected from tail vein of mice 7 d after the second dose and 14 d after the last dose and determined for specific IgG antibody levels by ELISA.Results The amantadine dimer was purified by thin layer chromatography(TLC)and identified by electrospray ionization-MS(ESI-MS)positive/negative ion mode.After two times of immunization,the antibody levels in sera at various dilutions of mice in R6A+RBD group were all higher than those of Ada+RBD group,while lower than those of Alu+RBD group.However,after three times of immunization,the antibody levels in sera at various dilutions of mice in R6A+RBD group were all significantly higher than those of Ada+RBD and Alu+RBD groups(each F > 30,each P < 0.000 1 and each P < 0.01).Conclusion Amantadine dimer adjuvant enhanced humoral immune response induced by SARS-CoV-2 protein vaccine in mice with good adjuvant effect,which may be used as an alternative adjuvant.This strategy based on existing drug transformation provided a new idea for the development of novel adjuvants.

Construction of "Internet + nursing" quality evaluation index system based on the perspective of service objects / 中国实用护理杂志

Objective:To construct a set of quality evaluation index system of "Internet + nursing" with clear evaluation criteria with the service objects as the evaluation subject and Service Quality model as the theoretical framework, and to provide an objective basis for the quality evaluation of "Internet + nursing".Methods:CNKI, Wanfang databases, VIP, PubMed, and Web of Science were searched from establishment to November 2021. The article determined the quality evaluation items of "Internet + nursing" based on the perspective of service objects by means of literature analysis, field research and Delphi expert inquiries. Analytic hierarchy process was used to confirm the weight of each indicator.Results:In this study, 22 experts were consulted for 2 rounds.The effective recovery rate of the two rounds of questionnaires were 100%. The judgment basis coefficient was 0.936, familiarity coefficient was 0.945 and authority coefficient was 0.941; in the first round, the Kendall harmony coefficients assigned to the importance of the first and second level indicators were 0.187 and 0.122, and the Kendall harmony coefficients assigned to the feasibility were 0.183 and 0.125; in the second round, the Kendall harmony coefficients assigned to the importance of the first and second level indicators were 0.241 and 0.190, and the Kendall harmony coefficients assigned to the feasibility were 0.218 and 0.166 (both P<0.01). The final evaluation index system for the "Internet + nursing" based on the perspective of service objects included 7 first-class indicators and 33 second-class indicators. Conclusions:The "Internet + nursing" quality evaluation index system constructed in this study based on the perspective of service objects is scientific, reliable and practical, which can be used as an evaluation tool for the current "Internet + nursing" quality evaluation and strategy optimization.

Clinical Application of the Next Generation Sequencing for Molecular Classification in Endometrial Carcinomas / 中山大学学报(医学科学版)

ObjectiveTo explore the clinical application of molecular classification in endometrial cancers with the next generation sequencing （NGS）. MethodsTotally 112 cases of endometrial carcinoma diagnosed by pathology in The Sun Yat-sen University Cancer Center were collected. All of them were tested by hybridized-capture second-generation sequencing based on 1，021 gene panel. The molecular variation spectrum of each subtype and its relationship between the clinicopathological features were analyzed. ResultsThe cases were distributed as follows： 8 （7.1%） POLE mutation， 34 （30.4%） mismatch repair deficient， 26 （23.2%） TP53 mutation， 44 （39.3%） non-specific molecular profile. The median tumor mutation burden was respectively 252.0， 38.4， 5.8 and 5.4 Muts/Mb. There were no significantly differences among four subtypes in clinicopathological features such as age， histological grade， lymph node metastasis and clinical stage. PTEN （75.5%）， PIK3CA （66.7%）， ARID1A （55.9%）， TP53 （40.2%）， NF1 （29.4%） were the most common mutations in endometrial cancers. ConclusionsThe utilization of NGS in endometrial cancers can simultaneously identify molecular subgroups， screen Lynch syndrome and obtain molecular variation spectrum， which can provide guidance for immunotherapy and targeted therapy， contribute to further accumulation and exploration of molecular genetic characteristics.

Metagenomic data-analysis reveals enrichment of lipopolysaccharide synthesis in the gut microbiota of atrial fibrillation patients / 中华心血管病杂志

Objective: To investigate the functional changes of key gut microbiota (GM) that produce lipopolysaccharide (LPS) in atrial fibrillation (AF) patients and to explore their potential role in the pathogenesis of AF. Methods: This was a prospective cross-sectional study. Patients with AF admitted to Beijing Chaoyang Hospital of Capital Medical University were enrolled from March 2016 to December 2018. Subjects with matched genetic backgrounds undergoing physical examination during the same period were selected as controls. Clinical baseline data and fecal samples were collected. Bacterial DNA was extracted and metagenomic sequencing was performed by using Illumina Novaseq. Based on metagenomic data, the relative abundances of KEGG Orthology (KO), enzymatic genes and species that harbored enzymatic genes were acquired. The key features were selected via the least absolute shrinkage and selection operator (LASSO) analysis. The role of GM-derived LPS biosynthetic feature in the development of AF was assessed by receiver operating characteristic (ROC) curve, partial least squares structural equation modeling (PLS-SEM) and logistic regression analysis. Results: Fifty nonvalvular AF patients (mean age: 66.0 (57.0, 71.3), 32 males(64%)) were enrolled as AF group. Fifty individuals (mean age 55.0 (50.5, 57.5), 41 males(82%)) were recruited as controls. Compared with the controls, AF patients showed a marked difference in the GM genes underlying LPS-biosynthesis, including 20 potential LPS-synthesis KO, 7 LPS-biosynthesis enzymatic genes and 89 species that were assigned as taxa harbored nine LPS-enzymatic genes. LASSO regression analysis showed that 5 KO, 3 enzymatic genes and 9 species could be selected to construct the KO, enzyme and species scoring system. Genes enriched in AF group included 2 KO (K02851 and K00972), 3 enzymatic genes (LpxH, LpxC and LpxK) and 7 species (Intestinibacter bartlettii、Ruminococcus sp. JC304、Coprococcus catus、uncultured Eubacterium sp.、Eubacterium sp. CAG:251、Anaerostipes hadrus、Dorea longicatena). ROC curve analysis revealed the predictive capacity of differential GM-derived LPS signatures to distinguish AF patients in terms of above KO, enzymatic and species scores: area under curve (AUC)=0.957, 95%CI: 0.918-0.995, AUC=0.940, 95%CI 0.889-0.991, AUC=0.972, 95%CI 0.948-0.997. PLS-SEM showed that changes in lipopolysaccharide-producing bacteria could be involved in the pathogenesis of AF. The key KO mediated 35.17% of the total effect of key bacteria on AF. After incorporating the clinical factors of AF, the KO score was positively associated with the significantly increased risk of AF (OR<0.001, 95%CI:<0.001-0.021, P<0.001). Conclusion: Microbes involved in LPS synthesis are enriched in the gut of AF patients, accompanied with up-regulated LPS synthesis function by encoding the LPS-enzymatic biosynthesis gene.

Research progress of nanopore sequencing in the clinic / 中华检验医学杂志

Because of the low throughput of current first-generation sequencing and the shortread length of second-generation sequencing, a new technology that overcomes the above shortcomings has emerged. The third-generation sequencing based on nanopore does not rely on the chain reaction of DNA polymerases and distinguishes bases by identifying electrical signals. It has broad application prospects and also faces more challenges. At present, it has many applications in detection of infectious pathogens, infectious disease prevention and control, genetic variation, and tumor diagnosis.

Bilateral adrenal hemorrhage caused by anticoagulant therapy after joint replacement surgery: One case report and literature review / 中华内分泌代谢杂志

Bilateral adrenal hemorrhage is a rare cause of primary adrenal insufficiency, and bilateral adrenal hemorrhage due to anticoagulant use is even rarer. We describe the case of a 62-year-old woman receiving post total knee arthroplasty anticoagulant therapy who presented fever, vomitting, stomachache, and severe fatigue on the 8th day. It was until 4 months later that the patient was finally diagnosed with adrenal insufficiency resuting from bilateral adrenal hemorrhage, her symptoms were relieved by glucocorticoid replacement therapy. In order to promote the awareness, diagnosis, and mangement of post-surgery anticoagulants induced bilateral adrenal hemorrhage, the clinical characteristics of the reported cases were summarized and analyzed.

Analysis of the characteristics of sarcopenia in elderly patients of different age groups / 中华老年医学杂志

Objective:To compare clinical characteristics and results on parameters for muscle measurement in elderly sarcopenia patients of different age groups.Methods:This was a cross-sectional study, and 404 outpatients aged 60 years and over were enrolled and received a long-term follow-up.According to the World Health Organization classification of the elderly, the subjects were divided into a youngest-old group, a middle old group and an oldest-old group; and according to the diagnostic criteria of the Asian Working Group for Sarcopenia in 2019, elderly people of all ages were divided into a sarcopenia group and a non-sarcopenia group.General clinical data were recorded, and the upper arm circumference, leg circumference, skeletal muscle index, grip strength and walking speed, the Short Physical Performance Battery and timed up and go were conducted.At the same time, the body composition was measured by multifrequency bioelectrical impedance analysis.Results:The subjects had a mean age of(85.24 ± 8.23)years, including 90(22.28%)youngest-old, 165(40.84%)middle old and 149(36.88%)oldest-old.123 patients(30.45%)were diagnosed with sarcopenia, with a prevalence of 27.78%(25)in the youngest-old group, 28.48%(47)in the middle old group and 34.23%(51)in the oldest-old group.Compared with those without sarcopenia, the basal metabolic rate in sarcopenia patients of all age groups decreased significantly, the levels of hemoglobin and albumin in youngest-oldsarcopenia patients decreased, the waist circumference and body mass index in middle old and oldest-old sarcopenia patients decreased, and the body fat rate in middle old sarcopenia patients increased.In the subjects with sarcopenia, the proportion of men in the oldest-old group was higher than in the other two groups.Compared with the youngest-old group, the levels of alanine aminotransferase and prealbumin decreased, serum creatinine increased, and skeletal muscle index values did not change in the middle old and longest-old groups, but handgrip strength(27.7±5.9 vs.23.2±6.9 vs.21.4±5.8, F=5.81, P=0.004), gait speed(0.8±0.2 vs.0.7±0.2 vs.0.5±0.2, F=11.88, P=0.000)and calf circumference(33.4±3.3 vs.15.9± 8.4 vs.31.5±3.1 vs.30.9±3.3, F=3.58, P=0.031)significantly decreased, and time up and go values(10.0±2.1 vs.15.9±8.4 vs.20.8±12.8, F=6.98, P=0.001)increased in the middle old and longest-old groups.Partial correlation analysis showed that age had a significant negative correlation with handgrip( r=-0.374, P=0.001), daily gait speed( r=-0.441, P=0.000), and calf circumference( r=-0.223, P=0.017), but a significant positive correlation with timed up and go( r=0.319, P=0.009). Conclusions:Compared with youngest-old sarcopenia patients, middle old patients show significant decline in muscle function, muscle strength and muscle mass in the lower limb.With increasing age, the impact of changes in muscle function, muscle strength and regional muscle mass on adverse events should receive heightened attention.

Clinical value of myoglobin in predicting multiple organ dysfunction in wasp sting patients / 中华急诊医学杂志

Objective:To investigate the clinical features and risk factors of multiple organ dysfunction syndrome (MODS) caused by wasp sting.Methods:A retrospective cohort study was conducted to collect the general data of wasp sting patients who had a clear history of wasp sting disease and clinical manifestations from June 2016 to December 2020 and were first diagnosed as wasp sting in hospital. Patients with hematological diseases, malignant tumors, severe liver and kidney dysfunction, cardiac insufficiency, and patients who had received hormone therapy before admission were excluded. Patients who were unable to obtain effective laboratory results due to hemolysis or other reasons within 48 h of admission were also excluded. The white blood cell count (WBC), neutrophil count (NEU), lymphocyte count (LYM), hemoglobin count (HB), myoglobin (Mb/MYO), activated partial thromboplastin time (APTT), albumin (ALB), K, Na, and Cl of the blood samples collected within 48 h after admission were recorded. Patients were divided into the MODS group and non-MODS group according to whether MODS occurred during hospitalization. Uni- and multivariate analysis were used to analyze the factors affecting the occurrence of MODS in wasp sting patients during hospitalization, and the receiver operating characteristic (ROC) curve was plotted to evaluate the predictive effect of myoglobin level on the occurrence of MODS in wasp sting patients during hospitalization.Results:Mb, WBC, NEU, APTT and serum potassium in the MODS group [3890.00 (1416.90-4057.00) ng/mL, (21.99 ± 8.18) × 10 9/L, (19.61 ± 7.33)× 10 9/L, (93.75 ± 45.77) s, and (4.99 ± 0.95) mmol/L] were significantly higher than those in the non-MODS group [73.50 (34.30-264.20) ng/mL, (13.40 ± 4.14)× 10 9/L, (11.18±4.73)× 10 9/L, (37.00 ± 17.16) s, and (4.05 ± 0.56) mmol/L] (all P < 0.05); blood chlorine and ALB [(101.50 (98.25-105.00) mmol/L and (35.36 ± 6.44) g/L)] were significantly lower than those in the non-MODS group [(105.00 (103.00-107.00) mmol/L and (40.71 ± 5.48) g/L)] (all P < 0.05). Multivariate logistic regression analysis showed that NEU ( OR = 0.729, 95% CI: 0.542~0.981), Mb ( OR = 0.999, 95% CI: 0.998~1.000), and APTT ( OR = 0.951, 95% CI: 0.921~0.982) were independent risk factors for MODS in wasp sting patients. ROC curve analysis showed that NEU, Mb and APTT could be used to evaluate the occurrence of MODS in wasp sting patients. Among them, Mb had the highest predictive value (AUC = 0.950, 95 % CI: 0.891~0.982). The optimal cutoff value of Mb for predicting the occurrence of MODS in wasp sting patients was 515.30 ng/mL, and the corresponding sensitivity and specificity were 90.62% and 87.23%, respectively. Conclusion:Mb is an independent risk factor for MODS in wasp sting patients, which can be used as a good predictor of MODS in wasp sting patients.

Significance of paying attention to extramural vascular invasion before operation for rectal cancer / 国际外科学杂志

Rectal cancer is one of the most common malignant tumors of digestive system. Accurate preoperative diagnosis plays an important role in surgeons′ decision on treatment plan. Extramural venous invasion (EMVI), as an important prognostic factor for predicting lymph node metastasis, distant metastasis and overall survival time of rectal cancer, has been paid more attention in recent years. It may even be used as a new staging system for guiding clinical treatment and formulate accurate preoperative staging. Preoperative MRI(mrEMVI) and postoperative pathology (pEMVI) is equal in the accuracy of assessment of EMVI. mrEMVI can be used as an important item for preoperative multi disciplinary team or clinicians to evaluate the patient stage and formulate individual treatment plan, and can also be used as an important evaluation for whether to perform surgery after preoperative chemoradiotherapy (CRT) and whether to prolong postoperative chemoradiotherapy time. EMVI should be taken as one of the necessary preoperative examinations for all the rectal cancer patients.

Comparison of the anti-androgen effects of cyproterone acetate and spironolactone on male-to-female transsexuals / 中华医学美学美容杂志

Objective:To compare the anti-androgenic effect of cyproterone acetate (CPA) and spironolactone (SPL) on male-to-female transsexuals.Methods:From January 2012 to September 2021, 185 male-to-female transsexuals (95 using CPA and 90 using SPL) who visited the Peking University Third Hospital and under stable medication for ≥3 months were enrolled, aged 16 to 40 (23±5) years. General information and laboratory indicators of the last visit were collected for a cross-sectional study.Results:The median doses of antiandrogens in the CPA group and SPL group were 25 mg/d and 80 mg/d, respectively. And the median dose of oral estradiol valerate in both groups was 2 mg/d. Testosterone level in the CPA group was significantly lower than the SPL group [0.7 (0.7-1.5) nmol/L vs. 13.2(6.7-18.4) nmol/L, U= 6 970.500, P<0.001]. The CPA group also had better subjective effects on testicular atrophy, erection decrease, body hair decrease, skin softening and figure feminization (all P<0.05). The prolactin level of CPA group was significantly higher than that of SPL group [21.5 (12.6-30.1) ng/ml vs. 11.9 (7.7-20.0) ng/ml, U= 2 053.500, P<0.001]. Conclusions:CPA has a more significant effect on lowering testosterone levels than SPL, and is better than SPL in terms of testicular atrophy, erection decrease, body hair decrease, skin softening and figure feminization, albeit with a potentially higher risk of hyperprolactinemia.

Factors related to sarcopenia in hospitalized elderly patients / 中华老年医学杂志

Objective:To investigate the prevalence of sarcopenia and associated factors in hospitalized elderly patients.Methods:This was a cross-sectional study.A total of 578 patients admitted to the Geriatrics Department of our hospital were consecutively recruited according to the admission criteria.Patients were divided into the sarcopenia group(n=202, 34.95%)and non-sarcopenia group(n=376, 65.05%)based on the diagnostic criteria(2014)of the Asian Working Group for Sarcopenia.Their clinical data and laboratory parameters were collected.All patients underwent comprehensive geriatric assessment.Results:(1)The detection rate of sarcopenia in these hospitalized patients was 34.95%.(2)Age, free thyroxine(FT4), and the prevalences of diabetes and osteoporosis and the incidence of falls in the past year were higher while body mass index(BMI), calf circumference, hemoglobin(Hb), albumin(ALB), triglycerides(TG), low-density cholesterol(LDL), glycosylated hemoglobin(HbA1c)and free triiodothyronine(FT3)were lower in sarcopenia patients than in non-sarcopenia patients, with statistical significance.Scores on the mini-mental state examination(MMSE)and activities of daily living(ADL)were lower in sarcopenia patients than in non-sarcopenia patients while scores on nutritional risk screening 2002(NRS2002)and the FRAIL scale were higher, all with statistical significance.(3)Binary logistic regression analysis revealed that osteoporosis and FRAIL score were risk factors for sarcopenia( OR=9.083 and 2.505, P<0.001)and BMI, calf circumference and ADL score were protective factors for sarcopenia( OR=0.735, 0.774 and 0.967, P<0.05). (4)ROC analysis showed that the areas under the curve for FRAIL score, BMI, calf circumference and ADL score were 0.832, 0.805, 0.841 and 0.812, respectively, with threshold values at 2.5 points, 23.52 kg/m 2, 32.5 cm and 92.5 points.(5)The chi-square test for sarcopenia screening using various related factors found osteoporosis, calf circumference <32.5 cm and ADL <90 points had higher sensitivity(0.787, 0.807, 0.817)while FRAIL ≥ 3 points and BMI <23.5 had slightly lower sensitivity(0.683, 0.708), with each related factor having a high negative detection rate(0.833-0.888). Conclusions:The prevalence of sarcopenia in hospitalized elderly is more than 1/3.Osteoporosis and FRAIL score are risk factors for sarcopenia.BMI, calf circumference and ADL score are protective factors for sarcopenia.All relevant factors have practical clinical value and can be used for preliminary screening of sarcopenia.

The role of integrin α4 in the anti-liver fibrosis effect of the sticky sugar amino acid extract of / 临床肝胆病杂志

Objective To investigate the mechanism of action of integrin α4 (ITGA4) in liver fibrosis based on the anti-liver fibrosis effect of sticky sugar amino acid (SSAA) in rats. Methods A rat model of liver fibrosis was induced by intraperitoneal injection of CCl 4 , and then colchicine and low-, middle-, and high-dose SSAA were used for intervention, with blank control group and SSAA group as control. After 12 weeks of experimental intervention, serum and liver samples were collected to measure the serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST), and HE staining and Sirius Red staining were used to observe the pathological conditions of liver tissue; quantitative real-time PCR was used to measure the transcriptional level of ITGA4, integrin β1 (ITGB1), transforming growth factor-β1 (TGFβ1), alpha-smooth muscle actin (α-SMA), and TIMP2 in liver tissue; Western blot was used to measure the relative protein expression levels of ITGA4, ITGB1, TGFβ1, α-SMA, MMP2, TIMP1, and TIMP2; immunohistochemistry was used to observe the protein expression of TGFβ1 and α-SMA. A one-way analysis of variance was used for comparison of continuous data between multiple groups, and the least significant difference t -test was used for comparison between two groups. Results There were significant increases in AST and ALT in the CCl 4 model group, and intervention with colchicine or low-, middle-, and high-dose SSAA reduced the levels of AST and ALT, with a significant difference between the CCl 4 model group and the other groups (all P < 0.05). HE staining and Sirius Red staining showed disordered structure of hepatic lobules and an increase in collagen fibers in the CCl 4 model group, and the structure of hepatic lobules was improved after intervention with colchicine or low-, middle-, and high-dose SSAA. The CCl 4 model group had significantly higher transcriptional levels of ITGA4, TGFβ1, α-SMA, and TIMP2 than the other groups, and there were significant reductions in the transcriptional levels of each factor after intervention with colchicine or SSAA, with a significant difference between the CCl 4 model group and the other groups (all P < 0.05). The CCl 4 model group had significantly higher protein expression levels of ITGA4, TGFβ1, α-SMA, TIMP2, and TIMP1 and a significantly lower protein expression level of MMP2 than the other groups, and intervention with colchicine or SSAA inhibited the expression of ITGA4, TGFβ1, α-SMA, TIMP2, and TIMP1 and promoted the expression of MMP2. Immunohistochemistry showed that the CCl 4 model group had significantly higher expression levels of TGFβ1 and α-SMA than the other groups, which was inhibited by intervention with colchicine or SSAA. The high-dose SSAA group had the most significant effect in reducing aminotransferases, improving lobular structure, and inhibiting the protein expression of liver fibrosis factors. Conclusion The high expression of ITGA4 in the liver is associated with the development of liver fibrosis, which is consistent with the increases in the expression of TGFβ1 and α-SMA. Inhibiting the expression of ITGA4 can provide more therapeutic targets for liver fibrosis and expand the anti-liver fibrosis mechanism of SSAA.

Optimization of CRISPR/Cas9-based multiplex base editing in Corynebacterium glutamicum / 生物工程学报

As a new CRISPR/Cas-derived genome engineering technology, base editing combines the target specificity of CRISPR/Cas and the catalytic activity of nucleobase deaminase to install point mutations at target loci without generating DSBs, requiring exogenous template, or depending on homologous recombination. Recently, researchers have developed a variety of base editing tools in the important industrial strain Corynebacterium glutamicum, and achieved simultaneous editing of two and three genes. However, the multiplex base editing based on CRISPR/Cas9 is still limited by the complexity of multiple sgRNAs, interference of repeated sequence and difficulty of target loci replacement. In this study, multiplex base editing in C. glutamicum was optimized by the following strategies. Firstly, the multiple sgRNA expression cassettes based on individual promoters/terminators was optimized. The target loci can be introduced and replaced rapidly by using a template plasmid and Golden Gate method, which also avoids the interference of repeated sequence. Although the multiple sgRNAs structure is still complicated, the editing efficiency of this strategy is the highest. Then, the multiple gRNA expression cassettes based on Type Ⅱ CRISPR crRNA arrays and tRNA processing were developed. The two strategies only require one single promoter and terminator, and greatly simplify the structure of the expression cassette. Although the editing efficiency has decreased, both methods are still applicable. Taken together, this study provides a powerful addition to the genome editing toolbox of C. glutamicum and facilitates genetic modification of this strain.

Clinical application of nanopore sequencing in detecting bacterial infections in lower respiratory tract / 中华检验医学杂志

Objective:To establish the method for detecting lower respiratory infections (LRIs) bacterialpathogens using nanopore sequencing, and evaluate the feasibility of this method.Methods:Bronchoalveolar lavage fluid (BALF) samples from 33 patients with LRIs who visited the Department of Respiratory and Critical Care Medicine of Beijing Hospital from July 2019 to September 2020 were collected.Nanopore 16S amplicon sequencing were performed on these samples. In order to evaluate the clinical value of the nanopore sequencing, χ 2 test was used to analyze the pathogen differences between the detection rate and pathogen types results found with using the nanopore 16S sequencing and the results found with bacterial culture. Results:The process and method of nanopore sequencing used in the detection of the LRIs pathogens were established. The pathogen detection rate of the 16S sequencing was higher than that of the traditional bacterial culture (75.8% [25/33], 45.5% [15/33], χ2=5.140, P<0.05). From the 25 positive samples found with nanopore 16S sequencing, 16 pathogens were detected, including Haemophilus parainfluenzae, Haemophilus influenzae, Streptococcus pneumoniae, Streptomonas maltophilia, Acinetobacter baumannii, and Acinetobacter junii, Staphylococcus aureus, Klebsiella pneumoniae, Enterococcus faecalis, Enterococcus gallinarum, Corynebacterium striatum, Mycobacterium paraintracellulare, Serratia marcescens, Achromobacter insuavis, Citrobacter murliniae and Mycoplasma pneumoniae. More than 6 pathogens were tested in clinical culture, including Haemophilus parainfluenzae, Acinetobacter baumannii, Pseudomonas aeruginosa, Staphylococcus aureus, Klebsiella pneumoniae and Streptomonas maltophilia (χ2=7.949, P<0.05). 16S sequencing aligned to species level sequences accounted for 80.0 (60.0, 86.0)% of the genus level. The results obtained by using16S sequencing and bacterial culture were consistent in 11 (33.3%) samples. Conclusions:Nanopore 16S amplicon sequencing can quickly identify pathogenic bacteria from BALF in LRIs patients. Nanopore 16S amplicon sequencing has a high detection rate, it can detect more pathogens than traditional bacterial culture, and it can also identify most bacteria to the species level. This technology is a very promising platform with broad application prospects.

Management of benign anastomotic stenosis after rectal cancer surgery / 国际外科学杂志

Objective:To discuss the occurrence, treatment and prevention of benign anastomotic stenosis after radical resection for rectal cancer.Methods:The clinical data of 63 patients with benign anastomotic stenosis from Jan. 2016 to Dec.2020 at Department of Gastrointestinal Surgery, Renji Hospital Shanghai Jiaotong University School of Medicine, were retrospectively analyzed, including general conditions, intraoperative conditions, postoperative complications, anastomotic stenosis, treatment process and outcome. The relationship between stenosis type and treatment and outcome were analyzed.The measurement data obeying normal distribution was expressed by ( Mean± SD), and the t test was used comparison between groups. The chi-square test was used comparison between enumeration data. Results:Of all the 63 patients, 22 (34.9%) cases presented with membranous stenosis, 30 (47.6%) cases with tubular stenosis, and 11 (17.5%) cases with diffused stenosis. Three of the 9 patients with high stenosis underwent balloon dilatation through endoscopy, 3 were placed with self-expandable metal stent and the rest 3 patients underwent resection and reconstruction of the anastomosis. All the 54 patients with low stenosis underwent digital anal expansion, and finally the effective rate was 53.7% (29/54). Endoscopic balloon dilatation was successfully performed in 8 cases, including 4 cases were placed metal stent throngh endoscopy. Eight patients underwent trans-anal stricturotomy. In 5 patients with low diffused stenosis, either ileostomy was preserved or permanent colostomy was performed due to failure to treatment. There were more male patients, protective ileostomy, anastomotic leakage and low stenosis in patients failed to treatment than in the cured patients ( P>0.05). However, all the 5 patients who failed to treatment were suffered from diffused stenosis, and the difference was statistically significant compared with those who were cured ( P<0.05). Conclusions:Postoperative anastomotic stricture after anterior rectectomy requires different treatment strategies according to the location and types of stricture. Endoscopic balloon dilatation is preferred for high stenosis, and metal stents can be placed optionally. Digital anal expansion is preferred for low anastomotic stenosis, and endoscopic or minimally invasive transanal surgery is feasible if digital anal expansion fails.

Dexmedetomidine alleviates hepatic ischemia-reperfusion injury by regulating MALAT1/miR-126-5p/HMGB1 axis / 生理学报

The aim of this study was to investigate the effects of dexmedetomidine (Dex) on hepatic ischemia/reperfusion injury (HIRI) and the underlying mechanism. The in vitro HIRI was induced by culturing HL-7702 cells, a human hepatocyte cell line, under 24 h of hypoxia and 12 h of reoxygenation. Quantitative real time PCR (qRT-PCR) and Western blot were performed to detect the expression levels of long non-coding RNA MALAT1, microRNA-126-5p (miR-126-5p) and high mobility group box-1 (HMGB1). Bioinformatics prediction and double luciferase assay were used to verify the targeting relationship between miR-126-5p and MALAT1, HMGB1. Reactive oxygen species (ROS), malondialdehyde (MDA) and ATP levels in culture medium were detected by corresponding kits. The results showed that Dex significantly reduced the levels of ROS and MDA, but increased the level of ATP in HL-7702 cells with HIRI. HIRI up-regulated the expression levels of MALAT1 and HMGB1, and down-regulated the level of miR-126-5p. Dex reversed these effects of HIRI. Furthermore, Dex inhibited HIRI-induced cellular apoptosis, whereas MALAT1 reversed the effect of Dex. This inhibitory effect of Dex could be restored by up-regulation of miR-126-5p. The results suggest that Dex protects hepatocytes from HIRI via regulating MALAT1/miR-126-5p/HMGB1 axis.

Anterior thalamic nuclei deep brain stimulation inhibits mossy fiber sprouting via 3',5'-cyclic adenosine monophosphate/protein kinase A signaling pathway in a chronic epileptic monkey model / 中华医学杂志(英文版)

BACKGROUND@#Anterior thalamic nuclei (ATN) deep brain stimulation (DBS) is an effective method of controlling epilepsy, especially temporal lobe epilepsy. Mossy fiber sprouting (MFS) plays an indispensable role in the pathogenesis and progression of epilepsy, but the effect of ATN-DBS on MFS in the chronic stage of epilepsy and the potential underlying mechanisms are unknown. This study aimed to investigate the effect of ATN-DBS on MFS, as well as potential signaling pathways by a kainic acid (KA)-induced epileptic model.@*METHODS@#Twenty-four rhesus monkeys were randomly assigned to control, epilepsy (EP), EP-sham-DBS, and EP-DBS groups. KA was injected to establish the chronic epileptic model. The left ATN was implanted with a DBS lead and stimulated for 8 weeks. Enzyme-linked immunosorbent assay, Western blotting, and immunofluorescence staining were used to evaluate MFS and levels of potential molecular mediators in the hippocampus. One-way analysis of variance, followed by the Tukey post hoc correction, was used to analyze the statistical significance of differences among multiple groups.@*RESULTS@#ATN-DBS is found to significantly reduce seizure frequency in the chronic stage of epilepsy. The number of ectopic granule cells was reduced in monkeys that received ATN stimulation (P < 0.0001). Levels of 3',5'-cyclic adenosine monophosphate (cAMP) and protein kinase A (PKA) in the hippocampus, together with Akt phosphorylation, were noticeably reduced in monkeys that received ATN stimulation (P = 0.0030 and P = 0.0001, respectively). ATN-DBS also significantly reduced MFS scores in the hippocampal dentate gyrus and CA3 sub-regions (all P < 0.0001).@*CONCLUSION@#ATN-DBS is shown to down-regulate the cAMP/PKA signaling pathway and Akt phosphorylation and to reduce the number of ectopic granule cells, which may be associated with the reduced MFS in chronic epilepsy. The study provides further insights into the mechanism by which ATN-DBS reduces epileptic seizures.