Connexin 43-modified bone marrow stromal cells reverse the imatinib resistance of K562 cells via Ca 2+ -dependent gap junction intercellular communication / 中华医学杂志(英文版)

BACKGROUND@#Imatinib mesylate (IM) resistance is an emerging problem for chronic myeloid leukemia (CML). Previous studies found that connexin 43 (Cx43) deficiency in the hematopoietic microenvironment (HM) protects minimal residual disease (MRD), but the mechanism remains unknown.@*METHODS@#Immunohistochemistry assays were employed to compare the expression of Cx43 and hypoxia-inducible factor 1α (HIF-1α) in bone marrow (BM) biopsies of CML patients and healthy donors. A coculture system of K562 cells and several Cx43-modified bone marrow stromal cells (BMSCs) was established under IM treatment. Proliferation, cell cycle, apoptosis, and other indicators of K562 cells in different groups were detected to investigate the function and possible mechanism of Cx43. We assessed the Ca 2+ -related pathway by Western blotting. Tumor-bearing models were also established to validate the causal role of Cx43 in reversing IM resistance.@*RESULTS@#Low levels of Cx43 in BMs were observed in CML patients, and Cx43 expression was negatively correlated with HIF-1α. We also observed that K562 cells cocultured with BMSCs transfected with adenovirus-short hairpin RNA of Cx43 (BMSCs-shCx43) had a lower apoptosis rate and that their cell cycle was blocked in G0/G1 phase, while the result was the opposite in the Cx43-overexpression setting. Cx43 mediates gap junction intercellular communication (GJIC) through direct contact, and Ca 2+ is the key factor mediating the downstream apoptotic pathway. In animal experiments, mice bearing K562, and BMSCs-Cx43 had the smallest tumor volume and spleen, which was consistent with the in vitro experiments.@*CONCLUSIONS@#Cx43 deficiency exists in CML patients, promoting the generation of MRD and inducing drug resistance. Enhancing Cx43 expression and GJIC function in the HM may be a novel strategy to reverse drug resistance and promote IM efficacy.

Dual-task treadmill training improves the motor functioning of children with bilateral spastic cerebral palsy / 中华物理医学与康复杂志

Objective:To explore any effect of the single- and dual-task treadmill training on the functioning of children with bilateral spastic cerebral palsy.Methods:Fifty children with bilateral spastic cerebral palsy were randomly divided into a single-task treadmill training group (the control group, n=25) and a dual-task treadmill training group (the observation group, n=25). All of the children also received routine rehabilitation training, and the control and observation groups also conducted single- and dual-task treadmill training in addition to the routine rehabilitation training, respectively. Before and after 2 months of treatment, each child′s gross motor functioning was quantified using sections D (standing) and E (walking, running and jumping) of the Gross Motor Function Measurement-88 (GMFM-88) instrument. Balance was quantified using the Pediatric Balance Scale (PBS) and walking mobility was quantified using a 1 minute walking test (1MWT). Modified and dual task Timed Up and Go (mTUG) tests and dual-task effects (DTE) tests were also administered. Results:There were no significant differences in average test scores between the two groups before the treatment. After the treatment significant improvement was observed in both groups. There was no significant difference between the two groups in terms of average GMFM-88, PBS and 1MWT scores, but significantly greater improvement was observed in the average dual-task mTUG and DTE results of the observation group.Conclusion:Both single- and dual-task treadmill training are effective supplements to routine rehabilitation training for children with bilateral spastic cerebral palsy. Dual-task treadmill training is more effective than the single-task version.

Pregnancy complicated with primary hyperparathyroidism: a report of 3 cases / 中华全科医师杂志

Clinical data of 3 pregnant women with primary hyperparathyroidism admitted in Jinan Maternal and Child Health Hospital from 2013 to 2021 were retrospectively reviewed. Hyperparathyroidism was diagnosed during pregnancy in all 3 cases. The clinical manifestations were non-specific, such as nausea, vomiting, anorexia, and fatigue. Patients also had gallbladder stones (1 case), gallbladder polyps (1 case), and renal parenchyma changes (2 cases). The hypercalcemia crisis occurred in all 3 cases, the blood Ca 2+ levels were 4.11 mmol/L, 2.92-3.82 mmol/L and 3.49-4.10 mmol/L, respectively; and preeclampsia developed in 2 cases. Sudden death occurred in case 1 who did not receive effective treatment during pregnancy; blood calcium was well controlled during pregnancy in case 2, and her neonate had hypocalcemia with good prognosis; case 3 underwent surgical treatment in early pregnancy, and then had missed abortion. Pathological examination revealed parathyroid tumors in all 3 cases.

Analysis of clinical features and pathogenic variants in a Chinese pedigree affected with congenital glycosylation disease / 中华医学遗传学杂志

OBJECTIVE@#To explore the clinical features and genetic basis for a Chinese pedigree diagnosed with congenital glycosylation disease (CGD).@*METHODS@#Clinical manifestations of two brothers were analyzed. Whole exome sequencing was carried out for the sib pair. Suspected variants were verified by Sanger sequencing.@*RESULTS@#Both the proband and her younger brother were found to carry compound heterozygous variants of the PMM2 gene, which included a known pathogenic mutation of c.395T>C (p.I132T) and a previously unreported c.448-1(delAG) in the 5' end of exon 6 of the gene.@*CONCLUSION@#The compound heterozygous variants of the PMM2 gene probably underlay the CGD in the sib pair.

Research progress on the relationship between the gut microbiota and geriatric diseases / 中华老年医学杂志

The human gut is populated by a complex community of microbiota that coexists with the host to maintain homeostasis.Macrogenomic and metabonomics analysis shows that the imbalance in gut microbiota composition and diversity, some bacterial components, metabolites and the signaling pathways mediated by them are involved in the occurrence and development of a variety of diseases, especially in the elderly.Intestinal microorganisms are one of the important factors affecting age-related diseases and can provide insights on treatment strategies for the elderly.This review focuses on the composition of the intestinal microbiota, interaction between the microbiota and the host, features of the gut microbiota in common geriatric diseases and pathogenic mechanisms.

Clinical Observation of Sacubitril Valsartan Sodium Tablets Combined with Bailing Capsules in the Treatment of Chronic Left Heart Failure with Renal Insufficiency / 中国药房

OBJECTIVE：To study the efficacy and safety of sacubitril valsartan sodium tablets combined with Bailing capsules in the treatment of chronic left heart failure with renal insufficiency ，and to provide reference for clinical drug use. METHODS ： Totally 96 patients with chronic left heart failure with renal insufficiency who sought medical care in our hospital from Nov. 2018 to Nov. 2019 were divided into group A ，B and C according to table of random numbers ，with 32 cases in each group. Group A received conventional heart failure treatment and and Bailing capsules （2 g each time ，3 times a day ）；group B received conventional heart failure treatment and Sacubitril valsartan sodium tablets （50 mg each time ，twice a day ）；group C was given with heart failure treatment and Sacubitril valsartan sodium tablets （50 mg each time ，twice a day ）and Bailing capsules （2 g each time，3 times a day ）. 3 groups received consecutive 6 months of treatment. Clinical response rates of 3 groups were compared. Left heart function indexes [left ventricular end systolic diameter （LVESD），left ventricular end-diastolic diameter （LVEDD），left ventricular ejection fraction （LVEF）] and serological indexes [interleukin 1（IL-1），IL-6，N terminal brain natriuretic peptide precursor，glomerular filtration rate （GFR）] were detected before and after treatment. The occurrence of ADR were observed and recorded. RESULTS ：During this study ，a total of 6 patients fell off ，and eventually 90 patients completed the study ，including 29 cases in group A ，30 cases in group B and 31 cases in group C. Before treatment ，there was no statistical significance in left heart function indexes or serological indexes among 3 groups（P＞ 0.05）. After 6 months of treatment ，clinical response rate of group C was significantly higher than those of group A and B 163.com （P＜0.05）. Compared with before treatment ， LVEDD， LVESD and serological indexes of 3 groups were decreased significantly after treatment （P＜0.05），while LVEF and GFR were increased significantly （P＜0.05）；the changes of above indexes （except for IL- 1 level in serum ） in group C were significantly better than group A and B ，the changes of above indexes in group B （except for GFR ）were significantly better than group A （P＜0.05）. No significant ADR were observed in 3 groups. CONCLUSIONS ：Sacubitril valsartan sodium tablets combined with Bailing capsules can significantly decrease the level of serum inflammation factors ，and improve cardiac and renal function in patients with chronic left heart failure with renal insufficiency ，with good safety.

Homogenization and optimization strategy for standard process of intensity-modulated radiotherapy for nasopharyngeal carcinoma / 中华放射肿瘤学杂志

Radiotherapy is the most common treatment for nasopharyngeal carcinoma, and the radiotherapy technique is essential for the prognosis of nasopharyngeal carcinoma. Due to the complexity of the structure of the intensity-modulated device and the accuracy of the clinical requirements of radiotherapy, it is inevitable that higher requirements will be imposed on the process of intensity-modulated radiotherapy. Currently, gaps exist in the radiotherapy equipment and personnel qualification among radiotherapy units, and thus the homogenization in the radiotherapy remains to be strengthened in China. With the application of radiotherapy information management system, digital medicine and artificial intelligence technologies in the field of radiotherapy, the original process fails to meet the application needs of the new precise radiotherapy technology. Therefore, this process is designed based on the existing radiotherapy procedures for nasopharyngeal carcinoma in combination with the latest developments in the field of radiotherapy, aiming to establish a novel standard process recommendation, ensuring the standardization and homogenization of radiotherapy and achieve the individualized intensity-modulated radiotherapy for nasopharyngeal carcinoma patients.

Analysis of Clinical Efficacy of Recombinant Human Brain Natriuretic Peptide Combined with Levosimendan in the Treatment of Acute Decompensated Heart Failure Complicated with Renal Insufficiency / 中国药房

OBJECTIVE：To observe the clinical effect and safety of recombinant human brain natriuretic peptide （rhBNP） combined with levosimendan in the treatment of acute decompensated heart failure （ADHF）complicated with renal insufficiency. METHODS：A total of 156 patients with ADHF complicated with renal insufficiency admitted to the Dept. of Cardiology in the Affiliated Hospital of Southwest Medical University during Jan.-Dec. 2019 were randomly divided into rhBNP group ，levosimendan group and combination group ，with 52 patients in each group. All patients received rountine treatment. On this basis ，rhBNP group was given rhBNP for injection [after 1.5 μg/kg intravenous pulse injection，intravenous dripping for 24 h with 0.007 5 μg（/ kg· min）]；leosimendan group was given Leosimendan injection 12.5 mg [intravenous dripping for 1 h with 6-12 μg（/ kg·min），then intravenous dripping for 23 h with 0.1 μg（/ kg·min）]. Combination group received drug combination according to the administration method of single drug group. Three groups received treatment for consecutive 7 d. Cardiac function indexes [heart rate （HR），left ventricular ejection fraction （LVEF），left ventricular end-diastolic diameter （LVEDD）]，mean arterial pressure （MAP），pulmonary capillary pressure （PCWP），renal function indexes [estimated glomerular filtration rate （eGFR），serum creatinine （Scr）]，serum levels of cystatin C （Cys-c）and amino-terminal brain natriuretic peptide precursor （NT-proBNP）were observed in 3 groups before and after treatment. Clinical efficacy and the occurrence of ADR were recorded. RESULTS ：Three cases withdrew from the study in rhBNP group and 1 case in levosimendan group ；152 cases completed the study. Before treatment ，there was no statistical significance in cardiac function indexes ，MAP，PCMP，renal function indexes or serum levels of Cys-C and NT-proBNP among 3 groups（P＞0.05）. After treatment ，the HP ，MAP，PCWP and serum level of NT-proBNP in 3 group as well as serum level of Cys-C in combination group were decreased significantly （P＜0.05）；the LVEF in 3 group as well as the eGFR and Scr level in levosimendan group and combination group were significantly increased （P＜0.05），compared with before treatment ；above indexes of combination group were significantly better than those of rhBNP group and levosimendan group （P＜0.05）. Total effective rate of combination group was 94.23％ ，which was significantly higher than those of rhBNP group （77.55％）and levosimendan group （76.47％）（P＜0.05）. There was no significant difference in the incidence of ADR among 3 groups（P＞ 0.05）. CONCLUSIONS ：rhBNP combined with levosimendan in the treatment of ADHF complicated with renal insufficiency can significantly increase the clinical efficacy ，and improve cardiac and renal function but don ’t increase the incidence of ADR.

The clinical observation of serum specific biomarkers in patients with chronic graft-versus-host disease / 中华血液学杂志

Objective@#Chronic graft-versus-host disease (cGVHD) is a major long-term complication after allogeneic hematopoietic stem cell transplantation (allo-HSCT) . It is important to study the changes of serum biomarkers expression in patients for early diagnosis and treatment.@*Methods@#The expression levels of five serum protein markers (IL-1b, IL-16, CXCL9, CCL19, CCL17) in patients with or without cGVHD after allo-HSCT were detected by liquid suspension microarray.@*Results@#Compared with the control group without cGVHD, the expression levels of CXCL9 and CCL17 in serum of patients with cGVHD were significantly increased (P<0.05) . CCL17 was correlated with the severity of cGVHD (P<0.001) . CXCL9 was significantly increased in the serum of patients with skin lesion (P<0.01) , and CCL17 was significantly expressed in cGVHD patients with liver as the target organ (P<0.01) .@*Conclusion@#The combination of CXCL9 and CCL17 can be used as serum biomarkers of cGVHD, which has certain reference value in assisting the diagnosis and evaluation of cGVHD severity.

Preliminary study of sequential multi-modality adjuvant chemotherapy and radiation for advanced endometrial cancer / 中华妇产科杂志

Objective To evaluate the clinical outcomes and feasibility of multi-modality adjuvant chemotherapy and radiation, which was conducted as postoperative chemotherapy, radiation, and consolidation chemotherapy (CRC) mode for the treatment of advanced endometrial cancer. Methods A retrospective analysis of 124 patients with International Federation of Gynecology and Obstetrics (FIGO) stages Ⅲ and Ⅳ endometrial cancer from Jan. 2004 to Oct. 2012 was conducted in Peking University People′s Hospital and Beijing Obstetrics and Gynecology Hospital. Inclusion criteria were comprehensive staging procedure including hysterectomy, bilateral salpingo-oophorectomy, and (or) selective pelvic aortic lymphadenectomy, and treatment with adjuvant chemotherapy and (or) radiation. The average age of these patients was (55.9 ± 8.4) years old (range from 23 to 79 years old). According to different postoperative adjuvant treatment modes, the patients were divided into CRC group, chemotherapy-radiotherapy (CR) group and single chemotherapy (C) group. The survival and side effects of the three groups were compared. Results (1) One hundred and twenty-four patients with advanced stage endometrial cancer were identified and received postoperative adjuvant therapies.Sixty-one (49.2％, 61/124) cases of them received postoperative CRC fashion, 19 (15.3％, 19/124) received postoperative CR and 44 (35.5％, 44/124) cases received C. The age, stage, grade and type of surgery of the three groups were not significantly different (all P>0.05);while, the pathology, chemotherapy cycles and chemotherapy regimens differed significantly (all P<0.05). (2) The progression-free survivals (PFS) of the patients with CRC, CR, and C group were (121±7), (68± 15), and (100±11) months, respectively. The 3-year PFS rates were 87.9％, 43.7％, and 61.4％, respectively. The 5-year PFS rates were 82.2％, 36.4％, and 61.4％, respectively. The above indicators were significantly higher in the CRC group than in the CR group (all P<0.01), and there was no difference between the CRC group and the C group (P=0.037). The overall survival (OS) of patients with CRC, CR, and C group were (128 ± 6), (80 ± 12), and (99 ± 10) months, respectively. The 3-year OS rates were 87.8％, 72.4％, and 67.1％, the 5-year OS rate were 84.2％, 54.3％, and 64.1％, respectively. The above indicators were significantly higher in the CRC group than those in the CR group and C group (all P<0.01). (3) There was no difference in the frequency of adverse effects either chemotherapy, such as severe bone suppression or radiotherapy;hepatotoxicity,blood transfusion, dose modifications;or cycle delays between the CRC, CR and C group (all P>0.05). (4) In the univariate analysis shown that, stage, the fashion of postoperative adjuvant therapy and type of surgery were risk factors for tumor progression in patients with advanced endometrial cancer (P<0.05). After adjusted for FIGO stage and type of surgery, the tumor progression hazard ratio (HR) was 3.931 (95％CI:1.734-8.914, P=0.001) for the CR group and 2.188 (95％CI:1.010-4.741, P=0.047) for the C group, compared to the CRC group. Conclusion Sequential CRC delivered in a"sandwich"fashion for the treatment in advanced endometrial cancer could significantly improve the 3-year and 5-year OS rates and have a similar adverse effect profile compared with other sequencing modalities.

A cross-sectional study of supervision in Shanghai mental health service industry / 中国心理卫生杂志

Objective:To understand the status of supervision of mental health service industry in Shanghai, and to provide reference for further strengthening the construction of mental health service system in Shanghai. Methods: In Shanghai, 107 mental health service agencies (including psychiatric institutions, non-specialist medical institutions, non-specialist enterprise and public institutions, social institutions) were selected from Shanghai psychological service industry association. The survey was conducted with the Questionnaire on Current Situation of Mental Health Professional Service Agencies in Shanghai (agency survey), including the "relevant information of agency" and the "management of the agency in the field of mental health services"(whether the agency had supervision requirements on the mental health service personnel, whether the agency organized supervision, the source of supervisors and payment method), and the Questionnaire on Current Situation of Mental Health Service Professionals in Shanghai (professional survey), including "basic information on demography "and" psychological professional work" (the number of hours of supervision in 2015, the cost of supervision, and the form of supervision and payment method). These two parts of questionnaire were selected and analyzed. Results: Agency survey results showed that 80％ of non-specialist enterprises had no supervision requirements for mental health service personnel, supervisor in non-specialist medical institutions were mainly from psychiatric institution (52.9％). The cost of supervision in most psychiatric institutions was paid by the agency (39.1％) and the cost of supervision in most public institutions was paid by professional (37％). The professional institutions under the jurisdiction of other departments within the unit required less supervision of the professional staff than the independent institutions (P ＜ 0.001). Professional survey results showed that the number of supervision hours of mental health service professional in social institutions was more than that in the other three types of institutions (P ＜0.001), of which 47.9％ chosen to pay the cost of supervision by themselves. The proportion of their own payment in social institutions was higher than that in the other three types of institutions (P ＜0.001). Conclusion:The social institutions in Shanghai have the best supervision of social institutions, followed by the supervision of the medical institutions. The supervision of non-medical enterprises and their personnel needs to be improved.

Development of mental health service agencies in Shanghai / 中国心理卫生杂志

Objective:To investigate the development status of mental health service of Shanghai mental health agencies,and to provide reference for further strengthening the standardized management of service agencies.Methods:In Shanghai,107 mental health service agencies (including psychiatric institutions,non-specialist medical institutions,non-specialist enterprise and public institutions,social institutions) were selected from Shanghai psychological service industry association.Each person in charge of these agencies was interviewed with an adapted 5 l-item questionnaire.A total of 89 valid questionnaires were collected.In this study,25 items of the questionnaire (belonging to the parts of agency information,mental health service situation and management of mental health service) were selected and analyzed.Results:All the surveyed agencies had been registered.The average age of these agencies for setting up mental health service was 8.9 years,and the average age of psychiatric institutions was the longest (18.6 years).Besides,psychiatric institutions attracted most of the clients (83.9％) in 2015.Among the professionals,only 32.5％ were full time,74.0％ were female,41.5％ were between 31 and 40 years old and 63.8％ were undergraduates.Psychiatrists,psychotherapists,counselors and psychometric person accounted for 78.0％ of the professionals in these agencies.Only 52.3％ of the agencies had full-time management personnel for mental health service.The most used method of assessing the quality of service and staff assessment was to obtain feedback from the client/family members (81.0％) and the assessment of the services (78.8％).Conclusion:The development of mental health service in Shanghai mental health agencies has been more normalized,but there is still a lack of full-time professionals.In addition,there is a lack of unified supervision and management mechanism for professionals.

Complex glycerol kinase deficiency: two case report and literature review / 临床儿科杂志

Objective To explore the clinical and genetic characteristics of complex glycerol kinase deficiency (GKD). Methods The clinical data of 2 cases of complex GKD were analyzed and the related literatures were reviewed. Results Both cases were male onset in neonatal period, and had hypocorticalism (hyponatremia, hyperkalemia, dehydration), hypercreatine kinasemia, and pseudotriglyceridemia. Gene detection suggested that there was gene deletion in chromosome Xp21 region. In the follow-up, one case had good control of the disease and one died of infection. Conclusions Complex GKD is an X-linked recessive hereditary disease. It is rare and complicated, and is easily misdiagnosed. Early diagnosis and treatment are beneficial to improve the prognosis.

Correlation analysis between glioblastoma perfusion imaging and isocitrate dehydrogenase 1 mutation status / 国际外科学杂志

Objective To investigate the value of perfusion imaging in predicting the status of isocitrate dehydrogenase 1 (IDH1) in glioblastoma.Methods Retrospectively studied 30 patients with glioblastoma multiforme with wild type IDH1 (IDHw) and 30 patients with mutant IDH1 (IDH1m) in Department of Neurosurgery,Shanxi Provincial People's Hospital from September 2014 to February 2016.Ktrans and Ve within the enhancing portion of each tumor were measured by using DCE-MRI data.rCBF and rCBV within the enhancing portion of each tumor were measured by using DSC-MRI.Four parameters were represented as (x) ± s,each of the 4 parameters was compared between patients with wild type IDH1 and mutant IDH1 by using the t-test.The performance in discriminating between the two entities was evaluated by using receiver operating characteristic analysis.Results Ktrans,Ve,rCBF and rCBV inside the enhancing lesion were significantly higher in patients with wild type IDH1 than in those with mutant IDH1.There was a statistically significant difference between IDH1w group and IDH1m group of rCBF value (P ＜ 0.05).The area under the curve for Ktrans,Ve,rCBF,and rCBV inside the enhancing lesion were 0.850,0.873,0.739 and 0.772,respectively.The value of rCBF value of the Ktrans value Ve value of IDH1w was significantly higher than that of IDH1m(P ＜0.05) in the value of the Ktrans value Ve value of rCBF,the value of the AUC value in rCBV was not significantly different with the combination of the 4 parameters of the diagnostic performance (AUC =0.915).Conclusions Ktrans,Ve,rCBF and rCBV calculated from MRI are useful for predicting the IDH1 mutation status.This method is not only for the classification of brain glioblastoma diagnosis has important value,and in glioblastoma classification is of important value in the preoperative noninvasive evaluation,and it has reference significance for predicting the prognosis of patients.

Effect of rhynchophylline on the mRNA expression of excitatory amino acid transporter 2 and NMDA receptor 2B after oxygen-glucose deprivation in cultured rat hippocampal astrocytes / 国际脑血管病杂志

Objective To investigate the effect of rhynchophylline on mRNA expression of excitatory amino acid transporter 2 (EAAT2 ) and N-methyl-D-aspartic acid receptor 2B (NR2B) after astrocyte oxygen-glucose deprivation. Methods The subcultured third generation astrocytes from the hippocampus were inoculated into 6-well plates, and they were divided into blank control group, hypoxia-ischemia group,low-dose rhynchophylline group (0. 02 mg/ml) and high-dose rhynchophylline group (0. 2 mg/ml) after the cells were attached to the wall and grew out protrusion. The total RNAs in each group were extracted.Real-time fluorescence quantitative polymerase chain reaction was used to detect the expression levels of EAAT2 and NR2B mRNA in astrocytes of each group. Results Compared with the blank control group, the expression levels of NR2B and EAAT2 mRNA in astrocytes of the ischemia-hypoxia group were significantly higher (all P < 0. 05 ). The expression levels of NR2B and EAAT2 mRNA in the low-dose rhynchophylline group were lower than those in ischemia-hypoxia group, but there was no significant difference. The expression levels of NR2B and EAAT2 mRNA in the high-dose rhynchophylline group were significantly lower than the ischemia-hypoxia group and the low-dose rhynchophylline group (all P < 0. 05).Conclusion The expression of EAAT2 and NR2B mRNA in astrocytes of hippocampus cultured in vitro was significantly increased after ischemia and hypoxia, and rhynchophylline intervention could significantly reduce its expression in a concentration dependent manner.

Targeted elimination of mutant mitochondrial DNA in MELAS-iPSCs by mitoTALENs

Mitochondrial diseases are maternally inherited heterogeneous disorders that are primarily caused by mitochondrial DNA (mtDNA) mutations. Depending on the ratio of mutant to wild-type mtDNA, known as heteroplasmy, mitochondrial defects can result in a wide spectrum of clinical manifestations. Mitochondria-targeted endonucleases provide an alternative avenue for treating mitochondrial disorders via targeted destruction of the mutant mtDNA and induction of heteroplasmic shifting. Here, we generated mitochondrial disease patient-specific induced pluripotent stem cells (MiPSCs) that harbored a high proportion of m.3243A>G mtDNA mutations and caused mitochondrial encephalomyopathy and stroke-like episodes (MELAS). We engineered mitochondrial-targeted transcription activator-like effector nucleases (mitoTALENs) and successfully eliminated the m.3243A>G mutation in MiPSCs. Off-target mutagenesis was not detected in the targeted MiPSC clones. Utilizing a dual fluorescence iPSC reporter cell line expressing a 3243G mutant mtDNA sequence in the nuclear genome, mitoTALENs displayed a significantly limited ability to target the nuclear genome compared with nuclear-localized TALENs. Moreover, genetically rescued MiPSCs displayed normal mitochondrial respiration and energy production. Moreover, neuronal progenitor cells differentiated from the rescued MiPSCs also demonstrated normal metabolic profiles. Furthermore, we successfully achieved reduction in the human m.3243A>G mtDNA mutation in porcine oocytes via injection of mitoTALEN mRNA. Our study shows the great potential for using mitoTALENs for specific targeting of mutant mtDNA both in iPSCs and mammalian oocytes, which not only provides a new avenue for studying mitochondrial biology and disease but also suggests a potential therapeutic approach for the treatment of mitochondrial disease, as well as the prevention of germline transmission of mutant mtDNA.

Comparative study of endoscopic ultrasonography and CT in the diagnosis of patients with pancreatic and periampullary tumors / 中国基层医药

Objective To evaluate the clinical value of endoscopic ultrasonography and CT in the diagnosis of patients with pancreatic and periampullary tumors.Methods From February 2010 to January 2016,89 patients with suspected pancreatic and periampullary tumors in Yinzhou Hospital Affiliated to Medical College of Ningbo University were selected in this study.Endoscopic ultrasonography and CT were performed in 89 patients.Endoscopic ultrasonography and CT examination results were compared with surgical and pathological results ,and the sensitivity, specificity and accuracy of the two methods were recorded and compared.Results By pathological examination , 69 cases were diagnosed as pancreatic and periampullary tumors.The sensitivity of endoscopic ultrasonography was 95.65%(66/69),the specificity was 95.00%(19/20),and the accuracy was 95.51%(85/89).The sensitivity of CT diagnosis was 85.51%(59/69),and the specificity was 90.00%(18/20),and the accuracy was 86.52%(77/89).The sensitivity and accuracy of endoscopic ultrasonography were significantly higher than those of CT ,and the differences were statistically significant (χ2＝4.161,4.395,P ＝0.041,0.036).And there was no statistically significant difference in the specificity of endoscopic ultrasonography and CT (χ2＝0.360,P＝0.548).Conclusion Endoscopic ultrasonography and CT examination have high specificity in the diagnosis of patients with pancreatic and periampullary tumors,however,the sensitivity and accuracy of endoscopic ultrasonography are higher than CT ,the clinical appropriate diagnostic methods can be chosen according to the actual condition of the patients .

Study on impairment of verbal fluency in patients with castration-resistant prostate cancer after chemotherapy / 中华行为医学与脑科学杂志

Objective To investigate the changes of verbal fluency test(VFT) in patients with castration resistant prostate cancer(CRPC) after chemotherapy.Methods 61 patients with CRPC were recruited.Thirty-one of those patients voluntarily accepted chemotherapy as CT group.The other thirty patients who did not received chemotherapy served as control group.CT group underwent VFT assessments at 3 time points:pre-chemotherapy (T1),during-chemotherapy (T2) and post-chemotherapy (T3).Patients in the control group were also assessed at the same time.Results (1) In phonetic fluency tasks,the interaction of time and group (time×group) between the CT group and control group was insignificant in the aspects of word capacity,cluster size,mean cluster size and phonetic switches(P＞0.05).The effect of time between the CT group and control group was significant difference in word capacity,cluster size,mean cluster size and phonetic switches(P＜0.05),but the effect of group was not significant(P＞0.05).CT group and control group both produced more words at T2 and T3 compared with that at T1 (P＜0.05).CT group had a lower score in cluster size and average cluster size than that of the control group at T3 ((6.17 ± 2.71) vs (7.64 ± 2.36),(3.14±0.78)vs (4.28± 1.53),P＜0.05).(2)In semantic fluency tasks,the interaction of time and group (time×group) between the CT group and control group was insignificant in the aspects of word capacity,cluster size,mean cluster size and semantic switches(P＞0.05).The effect of time between the CT group and control group was significantly different in word capacity,cluster size and semantic switches(P＜0.05),but the effects of groups and the effect of time in mean cluster were not significantly different(P＞0.05).The control group produced more words at T2 and T3 than T1 (P＜0.05).The CT group had a smaller cluster size at T3 than T1 and T2,and it was aslo smaller than control Group(P＜0.05).CT group had smaller cluster size at T3 than T2 ((5.47± 1.28) vs (7.15± 1.49),P＜ 0.05).At T3,the semantic switches of the CT group were more than T1,but less than control Group((20.81 ±3.63) vs (19.36±4.84) vs (21.54±6.18),P＜ 0.05)).The semantic switches of CT group were less at T2 than T1 ((18.25±4.25) vs (19.36±4.84),(P＜0.05)).Conclusion Standard dose chemotherapy has no significant impairment on the verbal fluency of patients with CRPC.Chemotherapy does not induce the impairment of patients' phonetic fluency,but may have a negative influence on semantic fluency.

Calycosin activates the protein kinase c/NF-E2-related factor 2 pathway to attenuate the oxidative stress and apopotosis induced by streptozotocin in RIN-m5F cell / 中国医师进修杂志

Objective To study the effects and mechanism of calycosin on oxidative stress andβ-cell apoptosis induced by streptozotocin (STZ). Methods RIN-m5F cells were divided into 5 groups:control group, STZ group, STZⅠgroup, STZⅡgroup and STZⅢgroup. The control group did not receive any treatment, while streptozotocin was added to the final concentration of 10 mmol/L in STZ group, STZⅠgroup, STZⅡgroup and STZⅢgroup. After the incubation with STZ for 6 h, calycosin was added to a final concentration of 10, 50 and 100 μmol/L in STZⅠgroup, STZⅡgroup and STZⅢgroup respectively. The cell viability and apoptosis was detected by CCK-8, LDH, caspase 3 and Tunel assay. The intracellular oxidative stress was measured using mitochondrial membrane potential, DCFH-DA, SOD activity and malondialdehyde levels assay. RIN-m5F cells were divided into control group, calycosinⅠgroup, calycosinⅡgroup and calycosinⅢgroup, which were treated with different concentrations (0,10, 50 and 100 μmol/L, respectively) of calycosin. The expression of NF-E2-related factor 2(Nrf2) in RIN- m5F cells was detected by Western blot. The translocation of Nrf2 was detected by immunofluorescence. In RIN-m5F cells were divided intoⅢgroup andⅣgroup,Ⅳgroup was pre-treated with protein kinase C(PKC) inhibitor. The effects of calycosin on Nrf2 translocation, oxidative stress and apoptosis were also observed. Results STZ could induce the accumulation of reactive oxygen species and apoptosis in RIN-m5F cells. Calycosin did not affect normal RIN-m5F cells, whereas it reduced the oxidative stress and apoptosis induced by STZ in a dose-dependent manner. The expression of Nrf2 in RIN-m5F cells was not affected by calycosin, whereas it promoted the translocation of Nrf2 into nucleus. The ability of calycosin promoting Nrf2 translocation was decreased after PKC inhibitor treatment, and PKC inhibitor could also significantly attenuate the anti-oxidant and anti-apoptotic ability of calycosin. Conclusions This study shows that calycosin may play an anti-oxidative and anti-apoptotic role by activating PKC to promote Nrf2 translocation, which is expected to be used as a new clinical drug for the prevention and treatment of diabetes mellitus.

Calycosin activates the protein kinase c/NF-E2-related factor 2 pathway to attenuate the oxidative stress and apopotosis induced by streptozotocin in RIN-m5F cell / 中国医师进修杂志

Objective To study the effects and mechanism of calycosin on oxidative stress andβ-cell apoptosis induced by streptozotocin (STZ). Methods RIN-m5F cells were divided into 5 groups:control group, STZ group, STZⅠgroup, STZⅡgroup and STZⅢgroup. The control group did not receive any treatment, while streptozotocin was added to the final concentration of 10 mmol/L in STZ group, STZⅠgroup, STZⅡgroup and STZⅢgroup. After the incubation with STZ for 6 h, calycosin was added to a final concentration of 10, 50 and 100 μmol/L in STZⅠgroup, STZⅡgroup and STZⅢgroup respectively. The cell viability and apoptosis was detected by CCK-8, LDH, caspase 3 and Tunel assay. The intracellular oxidative stress was measured using mitochondrial membrane potential, DCFH-DA, SOD activity and malondialdehyde levels assay. RIN-m5F cells were divided into control group, calycosinⅠgroup, calycosinⅡgroup and calycosinⅢgroup, which were treated with different concentrations (0,10, 50 and 100 μmol/L, respectively) of calycosin. The expression of NF-E2-related factor 2(Nrf2) in RIN- m5F cells was detected by Western blot. The translocation of Nrf2 was detected by immunofluorescence. In RIN-m5F cells were divided intoⅢgroup andⅣgroup,Ⅳgroup was pre-treated with protein kinase C(PKC) inhibitor. The effects of calycosin on Nrf2 translocation, oxidative stress and apoptosis were also observed. Results STZ could induce the accumulation of reactive oxygen species and apoptosis in RIN-m5F cells. Calycosin did not affect normal RIN-m5F cells, whereas it reduced the oxidative stress and apoptosis induced by STZ in a dose-dependent manner. The expression of Nrf2 in RIN-m5F cells was not affected by calycosin, whereas it promoted the translocation of Nrf2 into nucleus. The ability of calycosin promoting Nrf2 translocation was decreased after PKC inhibitor treatment, and PKC inhibitor could also significantly attenuate the anti-oxidant and anti-apoptotic ability of calycosin. Conclusions This study shows that calycosin may play an anti-oxidative and anti-apoptotic role by activating PKC to promote Nrf2 translocation, which is expected to be used as a new clinical drug for the prevention and treatment of diabetes mellitus.