Analysis of intraoperative frozen section diagnosis of 1 263 pulmonary nodules / 中国胸心血管外科临床杂志

@#Objective    To explore the key points and difficulties of intraoperative frozen section diagnosis of pulmonary diseases. Methods    The intraoperative frozen section and postoperative paraffin section results of pulmonary nodule patients in Beijing Chaoyang Hospital, Capital Medical University from January 2021 to January 2022 were collected. The main causes of misdiagnosis in frozen section diagnosis were analyzed, and the main points of diagnosis and differential diagnosis were summarized. Results    According to the inclusion criteria, a total of 1 263 frozen section diagnosis results of 1 178 patients were included in the study, including 475 males and 703 females, with an average age of 58.7 (23-86) years. In 1 263 frozen section diagnosis results, the correct diagnosis rate was 95.65%, and the misdiagnosis rate was 4.35%. There were 55 misdiagnoses, including 18 (3.44%) invasive adenocarcinoma, 17 (5.82%) adenocarcinoma in situ, 7 (35.00%) mucinous adenocarcinoma, 4 (2.09%) minimally invasive adenocarcinoma, 3 (100.00%) IgG4 related diseases, 2 (66.67%) mucinous adenocarcinoma in situ, 1 (16.67%) atypical adenomatous hyperplasia, 1 (14.29%) sclerosing pulmonary cell tumor, 1 (33.33%) bronchiolar adenoma, and 1 (100.00%) papillary adenoma. Conclusion    Intraoperative frozen section diagnosis still has its limitations. Clinicians need to make a comprehensive judgment based on imaging examination and clinical experience.

Modification of C20 oxidase in tanshinone biosynthesis pathway / 中国中药杂志

Tanshinones are one of the main effective components of Salvia miltiorrhiza, which play important roles in the treatment of cardiovascular diseases. Microbial heterogony production of tanshinones can provide a large number of raw materials for the production of traditional Chinese medicine(TCM) preparations containing S. miltiorrhiza, reduce the extraction cost, and relieve the pressure of clinical medication. The biosynthetic pathway of tanshinones contains multiple P450 enzymes, and the catalytic element with high efficiency is the basis of microbial production of tanshinones. In this study, the protein modification of CYP76AK1, a key P450-C20 hydroxylase in tanshinone pathway, was researched. The protein modeling methods SWISS-MODEL, Robetta, and AlphaFold2 were used, and the protein model was analyzed to obtain the reliable protein structure. The semi-rational design of mutant protein was carried out by molecular docking and homologous alignment. The key amino acid sites affecting the oxidation activity of CYP76AK1 were identified by molecular docking. The function of the obtained mutations was studied with yeast expression system, and the CYP76AK1 mutations with continuous oxidation function to 11-hydroxysugiol were obtained. Four key amino acid sites that affected the oxidation acti-vity were analyzed, and the reliability of three protein modeling methods was analyzed according to the mutation results. The effective protein modification sites of CYP76AK1 were reported for the first time in this study, which provides a catalytic element for different oxidation activities at C20 site for the study of the synthetic biology of tanshinones and lays a foundation for the analysis of the conti-nuous oxidation mechanism of P450-C20 modification.

Current situation and policy suggestions of medical social work supervision in shanghai / 中华医院管理杂志

Objective:To investigate the current situation of medical social worker supervision in Shanghai, for reference to promote the high-quality development of medical social work.Methods:From June to July 2022, a questionnaire survey was conducted on the in-service medical social workers in all medical institutions with medical social work departments or posts in Shanghai. The questionnaire mainly included demographic information, current status of supervision implementation, and effectiveness of supervision. The data were analyzed descriptively with t test for comparison between groups and the Pearson test was for correlation analysis. Results:A total of 99 medical social workers were included in this study, 65 had received supervision, and medical institutions where 58 people located had established the supervision system. The average scores of actual and expected supervisory support received by medical social workers were 3.71 and 4.20 respectively, and the mean burnout level score was 32.91. The effect of establishing a supervision institutions on burnout was statistically significant ( P<0.05) and actual access to supervision support was negatively associated with burnout ( P<0.05). Conclusions:The overall situation of medical social work supervision in Shanghai was positive, but the demand for supervision was not fully satisfied; Supervision had a positive impact on reducing the burnout level. It is suggested that medical social work should further strengthen the cultivation of supervisory talents, promote the construction of supervisory systems, and improve the quality of supervision.

Advances in the pathogenesis and treatment of short stature in Noonan syndrome / 国际儿科学杂志

Noonan syndrome(NS)is an inherited disease involving multiple systems.The main clinical manifestations include distinctive facial features, short stature, heart defects, developmental delay and chest deformity.Short stature, reported in up to 70% of NS patients, is one of the main reasons NS patients seek medical treatment.The pathogenesis is associated with the up-regulation of RAS-mitogen activated protein kinase(RAS-MAPK)signal pathway.Further study is needed for some further specific mechanisms.Recombinant human growth hormone(rhGH)therapy has been approved for NS patients with short stature and has achieved a good therapeutic effect.However, the knowledge of drug dosage, influencing factors, long-term efficacy and risk of rhGH treatment is still insufficient.This paper reviews the pathogenesis and treatment of short stature in NS, providing help for the treatment and management of the disease.

Clinical features of post-neurosurgical bacterial meningitis in children / 中华儿科杂志

Objective: To understand the characteristics of bacterial meningitis after pediatric neurosurgical procedures. Methods: This was a retrospective observational study. From January 2016 to December 2022, 64 children diagnosed with post-neurosurgical bacterial meningitis based on positive cerebrospinal fluid (CSF) culture in Department of Neurosurgery of Shanghai Children's Medical Center were selected as the study population. The clinical characteristics, onset time, routine biochemical indexes of cerebrospinal fluid before anti infection treatment, bacteriology characteristics and sensitivity to antibiotics of bacteria cultured from cerebrospinal fluid were analyzed. Based on the CSF culture results, the patients were divided into the Gram-positive bacteria infection group and the Gram-negative bacteria infection group. The clinical characteristics of the two groups were compared using t-tests or Wilcoxon rank-sum tests, and chi-square tests. Results: There were 64 children,42 boys and 22 girls, with onset age of 0.83 (0.50, 1.75) years. Seventy cases of post-neurosurgical bacterial meningitis occurred in the 64 children, of which 15 cases (21%) in spring, 23 cases (33%) in summer, 19 cases (27%) in autumn, and 13 cases (19%) in winter. The time of onset was 3.5 (1.0, 10.0) months after surgery; 15 cases (21%) occurred within the first month after the surgery, and 55 cases (79%) occurred after the first month. There were 38 cases (59%) showing obvious abnormal clinical manifestations, fever 36 cases (56%), vomiting 11 cases (17%). Forty-eight cases (69%) were caused by Gram-positive bacteria, with Staphylococcus epidermidis 24 cases; 22 cases (31%) were caused by Gram-negative bacteria, with Acinetobacter baumannii the prominent pathogen 7 cases. The Gram-positive bacterial infection was more common in summer than the Gram-negative bacterial infection (20 cases (42%) vs. 3 cases (14%), χ2=5.37, P=0.020), while the Gram-negative bacterial infection was more in autumn and within the first month after surgery than the Gram-positive bacterial infection (11 cases (50%) vs. 8 cases (17%), 15 cases (67%) vs. 5 cases (33%), χ2=8.48, 9.02; P=0.004, 0.003). Gram-positive bacteria resistant to vancomycin and Acinetobacter baumannii resistant to polymyxin were not found. However, Acinetobacter baumannii showed only 45% (10/22) susceptibility to carbapenem antibiotics. Conclusions: The clinical presentation of post-neurosurgical bacterial meningitis in children is atypical. Gram-positive bacteria are the main pathogens causing post-neurosurgical bacterial meningitis; Gram-negative bacterial meningitis are more likely to occur in autumn and within the first month after surgery. Acinetobacter baumannii has a high resistance rate to carbapenem antibiotics, which should be taken seriously.

Design and Verification of Human Metabolic Measurement System Based on STM32 / 中国医疗器械杂志

A high-precision human metabolic measurement system is designed. The system uses STM32F103 as the main control chip to acquire oxygen, carbon dioxide and flow signals to calculate four quantitative indicators: oxygen consumption(VO2), carbon dioxide production(VCO2), respiratory entropy(RQ) and resting energy metabolism(REE), and finally uses an upper computer to display the calculation results.In this paper, the signal acquisition circuit design was carried out for the oxygen sensor, carbon dioxide sensor and flow sensor, and the validity of the device was verified with the American machine MGCDiagnositcs using Bland-Altman analysis method, and the results showed that the four parameters of VO2,VCO2, RQ and REE of both devices fell in the agreement interval of more than 95%. The device thus provides accurate metabolic measurements and offers an effective tool for the field of general health and clinical nutrition support in China.

Evaluation of multiplex PCR for viral central nervous system infection in children / 中国小儿急救医学

Objective:To evaluate the clinical significance of molecular detection testing multiple pathogens in children with viral central nervous system infections.Methods:We retrospectively included 176 children who were suspected with central nervous system infection at Shanghai Children′s Medical Center from January 2017 to May 2021.Film Array Meningitis/Encephalitis Panel(FA-M/E) was used to test cerebrospinal fluid samples of these children.The results were analyzed compared with clinical symptoms and cerebrospinal fluid indices.Results:There were 34 samples with positive FA-M/E virus detection(19.32%, 34/176). Among the 34 samples, enterovirus was the most common pathogen(27 cases, 79.41%). In different combinations, the sensitivity and positive predictive value were all less than 90%.The median time for antiviral drugs used in FA-M/E virus-positive and negative children was 4.5(0, 8.5)d and 2.6(0, 2.0)d, respectively.The difference was statistically significant( P<0.05). Conclusion:Molecular tests of multiple pathogens can quickly and sensitively detect pathogens.It can improve the efficacy of clinical diagnosis of viral central nervous system infection.

Clinical analysis of 7 cases of community-acquired Novel Coronavirus Omicron variant infection in neonates / 中国新生儿科杂志

Objective:To study the clinical characteristics of neonatal community-acquired Novel Coronavirus (COVID-19) Omicron variant infection.Methods:From March 30 to May 15, 2022, the epidemiological characteristics, clinical manifestations and outcomes of neonatal cases of community-acquired COVID-19 Omicron variant infection admitted to the isolation ward of our hospital were analyzed.Results:A total of 7 neonates infected with community-acquired COVID-19 Omicron variant were treated, including 3 males and 4 females. All of them were term infants with clear epidemiological exposure history. The infection was originated from caregivers of close contact (parents or babysitters). The main clinical symptoms was upper respiratory tract infection, including fever (6 cases), nasal congestion (6 cases), cough (5 cases), runny nose (2 cases), poor appetite (2 cases) and diarrhea (1 case). On admission, no abnormalities were found in blood routine examination and C-reactive protein (CRP). All but one case had normal serum amyloid A (SAA). No obvious abnormalities were found on chest X-ray. All patients were isolated in single-patient rooms after admission. They received standard symptomatic treatment and regular nucleic acid tests. The first negative nucleic acid results came on median 17 d(8~26 d) after the onset of the disease. The patients were discharged after two consecutive (24 h apart) nucleic acid tests with CT value ≥35 and continued health-monitor at home. On discharge, 5 patients had nasal congestion and 2 of them had cough. During the follow-up 4~6 weeks after discharge, all patients gradually recovered without positive nucleic acid results.Conclusions:All 7 neonates with community-acquired COVID-19 Omicron variant infection have epidemiological exposure history. The main clinical symptoms are long-lasting upper respiratory tract infections. It takes a relatively long time for the nucleic acid to turn negative, however, the overall short-term prognosis is good.

Pathogen and clinical characteristics of bacterial enteritis in a third class children′s hospital in Shanghai city from 2016 to 2020 / 中国小儿急救医学

Objective:To investigate the pathogens and drug resistance of bacterial enteritis in children, analyze the clinical characteristics of bacterial enteritis in children, and provide basis for clinical diagnosis and treatment.Methods:The fecal culture strain and drug sensitivity of patients with bacterial enteritis admitted to our hospital from January 2016 to December 2020 were analyzed and summarized, and the clinical characteristics of patients who were infected by Salmonella and Escherichia coli were compared.Results:There were a total of 173 patients, aged from 21 days to 15 years, with a median age of 2.00(1.10, 3.54)years.Bacterial enteritis was most likely to occur in summer and autumn, and the incidence rate was 40.5% and 29.5%, respectively.One hundreds and seventy-three strains of bacteria were cultured in feces, including 148 strains of Salmonella(85.5%), 18 strains of Escherichia coli(10.4%), five strains of Staphylococcus aureus and two strains of Shigella.One hundreds and one of 141 patients who were infected with Salmonella were detected for leukocytes of in feces(71.6%), and four of 16 patients with Escherichia coli were detected for leukocytes(25.0%). The difference was significant( χ2=14.1, P<0.001). Eighty-eight of 113 patients(77.9%) who were infected by Salmonella with increased CRP(CRP>8 mg/L)and the proportion in Escherichia coli infection cases was 6/13(46.2%). There was significant difference( χ2=4.63, P=0.03). The drug sensitivity of Salmonella and Escherichia coli was summarized.There was no carbapenem resistant strain cultured; The sensitivity to piperacillin/tazobactam and cefoperazone/sulbactam was higher than 85%; The sensitivity to cefepime, ceftazidimeand ceftriaxone was higher than 75%; The sensitivity to ampicillin was lower than 30%, and the sensitivity to quinolones was between 20%-40%. Conclusion:Children aged 1-3 years are prone to bacterial enteritis in summer and autumn.The most common pathogens causing bacterial enteritis are Salmonella and Escherichia coli.White blood cells are more easily detected in feces of patients with Salmonella infection, and the increase rate of C-reactive protein in peripheral blood is higher.Patients with bacterial enteritis are recommended to use the third-generation cephalosporins and aforementioned antibiotics and piperacillin/tazobactam for empirical treatment.The sensitivity to quinolones is reduced, and may not be suitable for clinical application.

Rapid detection of pneumocystis carinii infection in immunocompromised patients by loop mediated isothermal amplification technique / 中国小儿急救医学

Objective:To explore the value of detecting pneumocystis carini(PC)rapidly in immunocompromised patients by loop mediated isothermal amplification(LAMP).Methods:Respiratory tract specimens of immunocompromised children suspected of pneumocystis carinii pneumonia(PCP) at Shanghai Children′s Medical Center, School of Medicine, Shanghai Jiao Tong University were collected from May 2020 to May 2021.PCR and LAMP methods were used to detect PC.Firstly, LAMP primers of PC were synthetized according to the conserved region of PC gene, and the LAMP reaction system and reaction conditions were optimized to evaluate the sensitivity and specificity.Then, the results of pathogens were compared with those of PCR detection.Results:The established LAMP detection technology for PC had high specificity and super sensitivity.The detection results could be obtained within 1 hour.In 12 clinical samples, 10 cases were positive and 2 cases were negative, the coincidence rate of LAMP and PCR technique was 100%.Conclusion:LAMP can detect PC more rapidly and sensitively than PCR, and it can provide a good support for clinical rapid diagnosis of PCP.

Rapid detection of pneumocystis carinii infection in immunocompromised patients by loop mediated isothermal amplification technique / 中国小儿急救医学

Objective:To explore the value of detecting pneumocystis carini(PC)rapidly in immunocompromised patients by loop mediated isothermal amplification(LAMP).Methods:Respiratory tract specimens of immunocompromised children suspected of pneumocystis carinii pneumonia(PCP) at Shanghai Children′s Medical Center, School of Medicine, Shanghai Jiao Tong University were collected from May 2020 to May 2021.PCR and LAMP methods were used to detect PC.Firstly, LAMP primers of PC were synthetized according to the conserved region of PC gene, and the LAMP reaction system and reaction conditions were optimized to evaluate the sensitivity and specificity.Then, the results of pathogens were compared with those of PCR detection.Results:The established LAMP detection technology for PC had high specificity and super sensitivity.The detection results could be obtained within 1 hour.In 12 clinical samples, 10 cases were positive and 2 cases were negative, the coincidence rate of LAMP and PCR technique was 100%.Conclusion:LAMP can detect PC more rapidly and sensitively than PCR, and it can provide a good support for clinical rapid diagnosis of PCP.

Application of loop mediated isothermal amplification in etiological diagnosis of infectious diseases / 国际儿科学杂志

Infectious diseases are still the most important diseases in infants and young children.Due to young age and imperfect immune function, infectious diseases in children are characterized by "urgent" , "serious" and "complex pathogens" , which often lead to serious complications.Rapid and accurate diagnosis is particularly important for children.However, the existing pathogen detection methods are inefficiency, subject to experimental conditions and person, and impopularized in hospitals.The pathogen detection method based on loop mediated isothermal amplification presents high specificity, low-cost and simple operation and is suitable for promotion in primary medical units, which has great application value in the detection of pathogens of children.In addition, the combination of loop mediated isothermal with paper-based micro-gene chip, intelligent automation and digital system will make progress to clinical etiological diagnosis.

Porcine Circular RNA Insulin-like Growth Factor 1 Receptor Promotes Adipocyte Differentiation / 中国生物化学与分子生物学报

Circular RNA (circRNA), as a competitive endogenous RNA (ceRNA), plays an importantrole in the regulation of cell differentiation. The purpose of this study was to identify and analyze porcinecircular RNA insulin-like growth factor 1 receptor (circIGF1R), explore its expression patterns, construct a ceRNA regulatory network related to circIGF1R, and explore the regulation of its ectopicexpression on adipogenic differentiation of mouse mesenchymal stem cells (C3H10T1 / 2) effect. Forwardand reverse PCR, Sanger sequencing, RNase R enzyme digestion tests, and qRT-PCR were used toverify that circIGF1R is a circRNA formed by the second exon of insulin-like growth factor 1 receptor(IGF1R). It was expressed in all tissues of pigs, and its expression level increased with age in adiposetissues. miRDB, TargetScan and miRWalk online software were used to predict circIGF1R target genes. RNAhybrid software was used for binding site prediction. DAVID bioinformatics functional analysissoftware was used to perform GO and KEGG enrichment analysis on candidate target genes. Cytoscapesoftware was used to construct the ceRNA network diagram. Based on the gene expression correlation andpredicted target relationship, the GO and KEGG enrichment analysis was drawn and the ceRNA networkwas constructed; the dual luciferase reporter gene test was used, and we found that circIGF1R andFABP4 can bind to ssc (Sus scrofa chromosome) -miR-133a-5p. The circIGF1R overexpression vectorwas successfully constructed and expressed in C3H10T1/ 2 cells. It was found that after overexpression ofcircIGF1R, the expression of key adipogenic regulatory factors CEBPa, CEBPß, FABP4 and PPAR? increased significantly(P<0. 01), and the number of lipid droplets increased significantly. The results ofthis study show that circIGF1R exists in pig adipose tissues, and may positively regulate the adipogenicdifferentiation of C3H10T1/ 2 cells through the ceRNA mechanism, which lays a theoretical foundation forfurther research on circIGF1R regulating the adipogenic differentiation of pig precursor intramuscularadipocytes.

Comparison of clinical efficacy between arthroscopy-assisted and open reduction and internal fixation in the treatment of Schatzker type I-III tibial plateau fractures / 中华骨科杂志

Objective:To compare the clinical efficacy of arthroscopy-assisted and open reduction and internal fixation in the treatment of Schatzker type I-III tibial plateau fractures.Methods:The data of patients with Schatzker type I-III tibial plateau fractures who were treated from August 2017 to July 2019 were retrospectively analyzed. According to the treatment, the patients were divided into the arthroscopic-assisted minimally invasive reduction and internal fixation group (arthroscopy group) and the conventional open reduction and internal fixation group (incision group). In the arthroscopy group, there were 30 patients, 19 males and 11 females were included; the age was 45.13±7.12 years old (range, 29-60 years). Among them, 13 cases were Schatzker type I fractures, 14 cases were Schatzker type II fractures, and 3 cases were Schatzker type III fractures. In the incision group, there were 30 patients, 17 males and 13 females were included; the age was 43.53±7.79 years old (range, 31-58 years). Among them, 11 cases were Schatzker type I fractures, 15 cases were Schatzker type II fractures, and 4 cases were Schatzker type III fractures. The operation time, intraoperative blood loss, postoperative ambulation time, postoperative complete weight-bearing time and postoperative complications were recorded. The degree of knee joint swelling, knee flexion and extension range of motion and the American knee society knee score (AKS score) were compared between the arthroscopy group and the incision group.Results:Both groups were followed up. The follow-up time of the arthroscopy group were 10-18 months, with an average of 14 months; the follow-up time in the incision group were 12-18 months, with an average of 15 months. In the arthroscopy group, the operation time (87.60±9.20 min vs. 94.33±10.65 min), intraoperative blood loss (57.16±9.63 ml vs. 71.93±11.15 ml), postoperative ambulation time (5.13±1.28 d vs. 6.17±1.53 d) and postoperative complete weight-bearing time (12.83±1.68 weeks vs. 14.23±1.77 weeks) were superior to the incision group, and the differences were statistically significant ( t=2.62, 5.49, 2.83, 3.94; all P<0.05). The healing time was 13.33±1.37 weeks in the arthroscopy group and 14.86±1.63 weeks in the incision group, and the difference was statistically significant ( t=3.94, P<0.001). At 1 year after surgery, the range of flexion and extension of knee joint in the arthroscopy group was 116.77°±12.46°, which was better than that in the incision group, which was 109.13°±9.89°, and the difference was statistically significant ( t=2.63, P=0.011). The AKS score in the arthroscopy group was 164.57±11.16 points, and the score in the incision group was 149.53±14.77 points, and the difference was statistically significant ( t=4.45, P<0.001). There were no malunion or compartment syndrome in the arthroscopy group and the incision group. The total incidence of complications in the arthroscopy group was 13% (4/30), including 2 cases of poor wound healing, 2 of poor knee range of motion after operation. The total incidence of complications in the incision group was 23% (7/30), including 4 cases of poor wound healing, 1 of wound infection, 2 of poor knee range of motion after operation. And the difference between the two groups in complication incidence was not statistically significant (χ 2=1.00, P=0.317). Conclusion:Arthroscopic-assisted reduction and internal fixation in the treatment of Schatzker I-III tibial plateau fractures has the advantages of less trauma, less bleeding, early mobility, fewer complications, and better knee joint function, and there are no obvious arthroscopic-related complications, which is a safe and reliable treatment method.

Cloning and functional characterization of isopentenyl diphosphate isomerase genes from Panax vietnamensis var. fuscidiscus / 药学学报

Isopentenyl diphosphate isomerase (IDI) is a key enzyme in the regulation of triterpenes biosynthesis and plays an important role in ginsenoside biosynthesis. In this study, two IDI genes, PvfIDI1 (GenBank No. MZ736417) and PvfIDI2 (GenBank No. MZ736418) were cloned from Panax vietnamensis var. fuscidiscus. The open reading frame of both PvfIDI1 and PvfIDI2 was 924 bp encoding 307 amino acids. The molecular weights of PvfIDI1 and PvfIDI2 were 34.84 kDa and 34.66 kDa, respectively, with theoretical pIs of 6.01 and 5.66. Bioinformatic analysis indicated that PvfIDI1 and PvfIDI2 contained two conserved sequences: TNTCCSHPL and WGEHELDY. Phylogenetic analysis showed that PvfIDI1 and PvfIDI2 were closely related to Panax notoginseng IDI. Expression analysis showed that both PvfIDI1 and PvfIDI2 genes are expressed in root, rhizome, stem and leaf of P. vietnamensis var. fuscidiscus. However, PvfIDI1 is highly expressed in the rhizome and PvfIDI2 is highly expressed in the stem. PvfIDI1 and PvfIDI2 recombinant proteins were expressed in E. coli; a functional coloration experiment showed that PvfIDI1 and PvfIDI2 could promote the accumulation of lycopene, indicating that both PvfIDI1 and PvfIDI2 encode functional IDI enzymes. The cloning and functional studies on PvfIDI1 and PvfIDI2 provide a foundation for the further study of IDI and the regulation of ginsenoside biosynthesis in P. vietnamensis var. fuscidiscus.

MYOZ2 Promoted Adipogenic Differentiation of C3H10T1 / 2 Cells by Negatively Regulating the Expression of TCAP / 中国生物化学与分子生物学报

The objective of this study was to investigate the expression profile of the myozenin2 (MYOZ2) gene and elucidate its effect on adipogenic differentiation of C3H10T1 / 2 cells and its possible mechanism∙ The longissimus dorsi‚ subcutaneous fat and liver tissue was collected from 180-day-old Mashen pigs‚ 60-day-old ICR mice‚ 35-day-old Ross broiler and 12-month-old Small tail han sheep‚ and the expression profile of the MYOZ2 gene mRNA was detected∙ The results showed that the MYOZ2 gene has similar patterns of tissue expression in examined species‚ with the highest expression level in longissimus dorsi‚ and a small amount of expression in the subcutaneous fat and liver tissue∙ After the MYOZ2 gene was silenced in C3H10T1 / 2 cells‚ qRT-PCR results showed that the expression levels of key adipogenic genes PPARγ and FABP4 were significantly down-regulated compared with the control group (P < 0∙ 01) ; Western Blotting results showed that the PPARγ protein content was significantly decreased (P < 0∙ 05) ; Oil red O staining showed that the number of lipid droplets and the content of triglyceride were significantly decreased after silencing MYOZ2 (P < 0∙ 05) ∙ The expression of fatty acid metabolism related genes SCD‚ FASN‚ SREBP1‚ NR1H3‚ DGAT1‚ PNPLA2‚ HSL‚ CES1‚ CPT1 after MYOZ2 silencing were detected by qRT-PCR∙ The results showed that SCD‚ FASN‚ SREBP1‚ PNPLA2 and HSL were significantly down-regulated (P < 0∙ 01) ‚ NR1H3 was significantly reduced (P < 0∙ 05) ‚ DGAT1 expression was down-regulated but the difference was not significant‚ CES1 and CPT1 were significantly up-regulated (P < 0∙ 05) ∙ The STRING database was used to construct a MYOZ2-related protein interaction network map‚ and it was found that MYOZ2 may affect the adipogenic differentiation through the interaction of titin-cap (TCAP) and PPARγ∙ After silencing TCAP‚ qRT-PCR results showed that compared with the control group‚ the expression of key adipogenic genes PPARγ and FABP4 were significantly up-regulated (P < 0∙ 01) ; Western Blotting results showed that PPARγ protein was significantly increased (P< 0∙ 05) ; Oil red O staining showed that the number of lipid droplets and the content of triglyceride were significantly increased after TCAP silencing (P < 0∙ 05) ∙ qRT-PCR was used to detect the expression of TCAP after silencing MYOZ2‚ and the results showed that the expression of TCAP was significantly increased (P<0∙ 01) ∙ In summary‚ MYOZ2 was highly expressed in longissimus dorsi and lower expressed in subcutaneous fat and liver tissues∙ In addition‚ MYOZ2 may regulate the expression of key adipogenic genes PPARγ and FABP4 through the interaction of MYOZ2-TCAP -PPARγ‚ and to further regulate the expression of fatty acid metabolism related genes SCD‚ FASN‚ SREBP1‚ NR1H3‚ DGAT1‚ PNPLA2‚ HSL‚ CES1 and CPT1‚ thus playing an important role in the process of adipogenic differentiation∙

FNDC5 Regulates the Adipogenic Differentiation of C3H10T1/2 Cells by Inhibiting the Phosphorylation of ERK1/2 / 中国生物化学与分子生物学报

The aim of this study was to explore the regulatory mechanism of Type Ⅲ domain-containing protein5 (FNDC5) on adipogenic differentiation in C3H10T1/2 cells. qRT-PCR and Western blot were used to detect the expression of FNDC5 during adipogenic differentiation of C3H10T1/2 cells. The lentivirus-coated overexpression and interference vector of FNDC5 were constructed and transfected into C3H10T1/2 cells. qRT-PCR was used to detect the expression of the key genes of adipogenic differentiation. Oil red O staining was used to detect the formation of lipid droplets; Western blot was used to detect the content of ERK1/2 and ERK1/2 phosphorylated protein (P-ERK1/2). After 8 days of adipogenic differentiation of C3H10T1/2 cells, the expression of Fndc5 increased significantly. After overexpression of FNDC5 in C3H10T1/2 cells, the expression of key genes for adipogenic differentiation, including peroxisome proliferator-activated receptor-酌 (PPAR酌), CCAAT enhancer binding protein beta (C/EBP茁), fatty acid binding protein 4 (FABP4) and CCAAT enhancer binding protein alpha (C/EBPα), all decreased significantly. The content of lipid droplets and P-ERK1/2 also decreased significantly. On the contrary, after interference of FNDC5 in C3H10T1/2 cells, the expression of key genes for adipogenic differentiation, including PPARγ, C/EBP茁, FABP4 and C/EBPα were significantly increased. Meanwhile, the content of lipid droplets and P-ERK1/2 also increased significantly. This study found that FNDC5 can inhibit the adipogenic differentiation of C3H10T1/2 cells by inhibiting the phosphorylation level of ERK1/2, which can provide reference data for the mechanism of FNDC5 in regulating fat deposition.

Effect and Involved Mechanism of RSL3-induced Ferroptosis in Acute Leukemia Cells MOLM13 and Drug-resistant Cell Lines / 中国实验血液学杂志

OBJECTIVE@#To investigate the effect and involved mechanism of RSL3 on ferroptosis action in acute leukemia cells MOLM13 and its drug-resistant cells.@*METHODS@#After MOLM13 treated with RSL3, CCK-8 assay was performed to detect cell viability, flow cytometry was used to detect the reactive oxygen species (ROS) level of the cells, RT-qPCR and Western blot were used to detect the expression of glutathione peroxidase 4 (GPX4). After MOLM13/IDA and MOLM13/Ara-C, the drug-resistant cell lines were constructed, the ferroptosis induced by RSL3 was observed. Bone marrow samples were collected from patients with acute monocytic leukemia. RT-qPCR and Western blot were performed to detect the expression of related genes and proteins involved in ferroptosis pathway.@*RESULTS@#RSL3 significantly inhibited the cell viability of MOLM13 and increased the intracellular ROS level, which were partially reversed by ferrostatin-1. The mRNA and protein expression of GPX4 decreased in MOLM13 treated with RSL3. RSL3 inhibited the viability of MOLM13/IDA and MOLM13/Ara-C cells more strongly than that of non-drug resistant cells, also increased the intracellular ROS level . The cytotoxic effects were partially reversed by ferrostatin-1. The mRNA and protein expressions of GPX4 in MOLM13/IDA and MOLM13/Ara-C cells were higher than those in non-drug resistant cells. The mRNA and protein levels of GPX4 in bone marrow of relapsed/refractory acute mononuclear leukemia patients were higher than those of ordinary acute mononuclear leukemia patients.@*CONCLUSION@#RSL3 can induce non-drug resistant cells MOLM13 ferroptosis by inhibiting GPX4 activity. MOLM13/IDA and MOLM13/Ara-C are more sensitive to RSL3 compared with non-drug resistant cells MOLM13, which may be caused by the differences in GPX4 expression. The expressions of GPX4 mRNA and protein in relapsed/refractory acute mononuclear leukemia are higher than those in ordinary acute mononuclear leukemia.

Chemical constituents from Clausena excavata and their inhibitory activities against proliferation of synoviocytes / 中国中药杂志

The chemical constituents from the stems and leaves of Clausena excavata were isolated and purified by column chromatography with silica gel, ODS, Sephadex LH-20 and RP-HPLC. The chemical structures of the isolated compounds were identified on the basis of physicochemical properties, spectroscopic analysis, as well as the comparisons with the data reported in literature. Nineteen compounds were isolated from the 90% ethanol extract of the stems and leaves of C. excavata, which were identified as methyl orsellinate(1), syringaresinol(2), lenisin A(3), scopoletin(4), osthenol(5), N-benzoyltyrarnine methyl ether(6), N-p-coumaroyltyramine(7), aurantiamide acetate(8), 1H-indole-3-carboxaldehyde(9), furostifoline(10), clausenalansine E(11), 3-formylcarbazole(12), clausine L(13), clausine E(14), methyl carbazole-3-carboxylate(15), glycosinin(16), murrayafoline A(17), clausine H(18) and 2,7-dihydroxy-3-formyl-1-(3'-methyl-2'-butenyl)carbazole(19). Among these isolated compounds, compounds 1-11 were isolated from C. excavata for the first time, and compounds 1, 2 and 10 were isolated from the genus Clausena for the first time. In addition, this study evaluated the anti-rheumatoid arthritis activities of compounds 1-19 by measuring their anti-proliferative effects on synoviocytes in vitro according to MTS method. Compounds 10-19 displayed remarkable anti-rheumatoid arthritis activities, which exhibited the inhibitory effects on the proliferation of MH7 A synovial fibroblast cells with the IC_(50) values ranging from(27.63±0.18) to(235.67±2.16) μmol·L~(-1).

Selection of optimal qRT-PCR reference genes for Aconitum vilmorinianum / 中国中药杂志

Screening suitable reference genes is the premise of quantitative Real-time PCR(qRT-PCR)for gene expression analysis. To provide stable reference genes for expression analysis of genes in Aconitum vilmorinianum, this study selected 19 candidate re-ference genes(ACT1, ACT2, ACT3, aTUB1, aTUB2, bTUB, 18S rRNA, UBQ, eIF2, eIF3, eIF4, eIF5, CYP, GAPDH1, GAPDH2, PP2A1, PP2A2, ACP, and EF1α) based on the transcriptome data of A. vilmorinianum. qRT-PCR was conducted to profile the expression of these genes in the root, stem, leaf, and flower of A. vilmorinianum. The Ct values showed that 18S rRNA with high expression level and GAPDH2 with large expression difference among organs were not suitable as the reference genes. NormFinder and geNorm showed similar results of the expression stability of the other candidate reference genes and demonstrated PP2A1, EF1α, and CYP as the highly stable ones. However, BestKeeper suggested EF1α, ACT3, and PP2A1 as the top stable genes. In view of the different results from different softwares, the geometric mean method was employed to analyze the expression stability of the candidate re-ference genes, the results of which indicated that PP2A1, EF1α, and ACT3 were the most stable. Based on the comprehensive analysis results of geNorm, NormFinder, BestKeeper, and geometric mean method, PP2A1 and EF1α presented the most stable expression in different organs of A. vilmorinianum. PP2A1 and EF1α were the superior reference genes for gene expression profiling in different organs of A. vilmorinianum.