Investigation and comparative study on death cognition and hospice care attitude of the graduate students of clinical medicine / 中华医学教育探索杂志

Objective:To investigate and analyze the status quo of death cognition and hospice care attitude of clinical medical graduate students, to compare the differences between the two groups, and to explore the correlation between death cognition and hospice care attitude, so as to provide the reference for the reform and construction of death cognition and hospice care education in medical colleges and universities in China.Methods:A survey was carried out on 496 doctoral and postgraduate students majoring in clinical medicine by using the "Questionnaire of General Sociology Survey", the Chinese version of the "Death Attitude Profile Revised Scale", and the Chinese version of the "Frommelt Attitudes Toward Care of the Dying Scale Form B (FATCOD-B)" to investigate the death cognition and hospice care attitude of 496 doctoral and postgraduate students majoring in clinical medicine, and classified and compared the survey results of the two groups. The results of this study were statistically analyzed by t-test, Pearson correlation analysis and other statistical methods. Results:A total of 469 valid questionnaires were recovered after excluding 27 unqualified questionnaires. Statistical analysis showed that the scores of each dimension in the death attitude description scale of doctoral students were ranked as follows: approach acceptance (4.28±0.53), neutral acceptance (3.99±0.41), death avoidance (2.74±0.63), fear of death (2.65±0.57) and escape acceptance (2.47±0.69) the scores of postgraduates were ranked as neutral acceptance (3.96±0.52), approach acceptance (2.84±0.61), fear of death (2.78±0.65), death escape (2.62±0.73), escape acceptance (2.39±0.77). At the same time, the scores of hospice care attitude in the doctoral group were higher than those in the master group [(110.63±8.96) vs. (106.78±6.52)], and the difference was statistically significant ( P<0.001). In addition, the scores of "fear of death" and the scores of hospice care attitudes were negatively correlated with the scores of doctoral and master students ( r=-0.25, r=-0.21), while the scores of hospice care attitudes were positively correlated with the scores of "neutral acceptance" in death cognition ( r=0.50, r=0.32). However, the hospice care attitude scores of doctoral students were negatively correlated with the "death avoidance" scores in death cognition ( r=-0.27). Conclusion:Doctoral and postgraduate students have a certain awareness of hospice care, while their death cognition and hospice care attitude still need to be improved and strengthened from sociology, psychology, ethics and other perspectives. The above will promote the harmony between doctors and patients while meeting the needs of an aging society in China.

Role of melatonin receptor 1B gene polymorphism and its effect on the regulation of glucose transport in gestational diabetes mellitus / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Melatonin receptor 1B (MT2, encoded by the MTNR1B gene), a high-affinity receptor for melatonin, is associated with glucose homeostasis including glucose uptake and transport. The rs10830963 variant in the MTNR1B gene is linked to glucose metabolism disorders including gestational diabetes mellitus (GDM); however, the relationship between MT2-mediated melatonin signaling and a high birth weight of GDM infants from maternal glucose abnormality remains poorly understood. This article aims to investigate the relationship between rs10830963 variants and GDM development, as well as the effects of MT2 receptor on glucose uptake and transport in trophoblasts. TaqMan-MGB (minor groove binder) probe quantitative real-time polymerase chain reaction (qPCR) assays were used for rs10930963 genotyping. MT2 expression in the placenta of GDM and normal pregnant women was detected by immunofluorescence, western blot, and qPCR. The relationship between MT2 and glucose transporters (GLUTs) or peroxisome proliferator-activated receptor γ (PPARγ) was established by western blot, and glucose consumption of trophoblasts was measured by a glucose assay kit. The results showed that the genotype and allele frequencies of rs10830963 were significantly different between GDM and normal pregnant women (P<0.05). The fasting, 1-h and 2-h plasma glucose levels of G-allele carriers were significantly higher than those of C-allele carriers (P<0.05). Besides, the protein and messenger RNA (mRNA) expression of MT2 in the placenta of GDM was significantly higher than that of normal pregnant women (P<0.05). Melatonin could stimulate glucose uptake and GLUT4 and PPARγ protein expression in trophoblasts, which could be attenuated by MT2 receptor knockdown. In conclusion, the rs10830963 variant was associated with an increased risk of GDM. The MT2 receptor is essential for melatonin to raise glucose uptake and transport, which may be mediated by PPARγ.

Effect of group mindfulness-based stress reduction therapy in managing comorbid depression in patients with stable chronic obstructive pulmonary disease / 四川精神卫生

BackgroundChronic obstructive pulmonary disease （COPD） is a common chronic respiratory disease， and patients with COPD often experience substantially emotional difficulties， such as anxiety and depression， all of which may cause serious detriment to the prognosis of patients. As a non-pharmacological intervention in clinical practice， group mindfulness-based stress reduction therapy （MBSR） is beginning to emerge， while has rarely been studied in COPD patients with concurrent emotional difficulties. ObjectiveTo evaluate the effects of group MBSR on depression， state of mindfulness and pulmonary function in stable COPD patients， so as to provide references for the application of group MBSR in patients with COPD. MethodsA total of 97 patients with stable COPD who were followed up in the Department of Respiratory and Critical Care Medicine of Mianyang Third People's Hospital from January to October 2019 were selected as the study objects， and they were assigned into study group （n=50） and control group （n=47） by random number table method. All individuals received routine medication therapy and an 8-week health education， based on this， participants in study group partook an 8-week intervention comprising group MBSR. At the baseline， 4 weeks and 8 weeks of intervention， participants were assessed with Self-rating Depression Scale （SDS）， Five Facet Mindfulness Questionnaire （FFMQ） and COPD Assessment Test （CAT）， as well as the pulmonary function testing. ResultsThere were 41 patients in study group and 42 cases in control group completed the study. The group * time interaction was interpreted as significant between two groups for SDS， FFMQ and CAT scores （F=54.858， 86.161， 69.862， P<0.01）. Baseline SDS， FFMQ and CAT scores of the two groups yielded no statistical difference between two groups （F=0.240， 0.052， 0.019， P>0.05）， while study group scored lower on SDS and CAT （F=12.900， 38.511， 7.797， 28.824， P<0.01） and higher on FFMQ （F=27.324， 82.412， P<0.01） than those of the control group after 4 and 8 weeks of intervention. With the prolongation of intervention time in study group， participants demonstrated an overall reduction in SDS and CAT scores （F=109.753， 124.144， P<0.01）， and an increase in FFMQ scores （F=228.194， P<0.01）. There were no between-group differences in forced expiratory volume in one second as percentage of predicted volume （FEV1%pred） after 4 and 8 weeks of intervention （F=0.104， P=0.748） ， and the within-group changes in FEV1%pred value over the intervention period in study group was not statistical （F=0.561， P=0.458）. ConclusionGroup MBSR may help relieve depressive symptoms， enhance mindfulness level， and alleviate clinical symptoms in stable COPD patients， but has no effect on pulmonary function. ［Funded by Mianyang Health and Health Commission Scientific Research Project （number， 201916）］

Echinococcus granulosus cyst fluid(EgCF) inhibits the migration and phagocytic function of mouse macrophages induced by LPS via inducing cytoskeletal rearrangement / 细胞与分子免疫学杂志

Objective To investigate the effect of Echinococcus granulosus cyst fluid(EgCF) on the cytoskeletal rearrangement and phagocytosis and the migration of macrophages induced by lipopolysaccharide(LPS). Methods Peritoneal macrophages of C57BL/6 mice were isolated and cultured in vitro, and divided into control group and LPS group and LPS combined with EgCF group. After 48 hours of treatment, filamentous actin (F-actin) changes were observed with rhodamine-labelled phalloidin staining and fluorescence microscopy; TranswellTM chamber was used to test cell migration ability and flow cytometry to test cell phagocytosis. After 1 hour of treatment, PI3K and AKT, phosphorylated AKT (p-AKT), Rac1, guanosine triphospho-Rac1 (GTP-Rac1), WASP and Arp2 protein expressions were detected with Western blot analysis. Results Compared with the control group, after LPS stimulation, macrophages were deformed significantly; pseudopodia increased; actin cytoskeleton increased and was more distributed in pseudopodia; the ability of migration and phagocytosis were significantly improved, and the expression of PI3K, p-AKT, GTP-Rac1, WASP and Arp2 proteins significantly increased. EgCF treatment caused cell shrinkage and disappearance of pseudopodia protrusions of LPS-activated cells, and led to the reduced phagocytic and migratory of cells; the protein expression of PI3K, p-AKT, GTP-Rac1, WASP and Arp2 decreased significantly compared with the LPS group. Conclusion LPS induces the migration and enhances phagocytosis of macrophages while EgCF inhibits these effects, which is related to actin cytoskeleton rearrangement.

Expression and clinical significance of Flap endonuclease 1 in acute myeloid leukemia / 肿瘤研究与临床

Objective:To investigate the expression level of flap endonuclease 1 (FEN1) in bone marrow mononuclear cells of patients with acute myeloid leukemia (AML) and its relationship with clinicopathologic features and therapeutic effect, so as to provide a new direction for disease monitoring and targeted therapy in AML patients.Methods:The data of 57 newly treated AML patients and 26 healthy individuals (the healthy control) from the First Clinical College of Guangdong Medical University and Fujian Medical University Union Hospital from November 2018 to December 2020 were retrospectively analyzed. Bone marrow samples of all subjects were collected. Quantitative real-time fluorescence polymerase chain reaction (qRT-PCR) was used to detect FEN1 mRNA expression in bone marrow mononuclear cells of all subjects. Bone marrow samples from 9 newly-diagnosed AML patients and 4 healthy controls were collected, and FEN1 protein expression level was detected by using Western blotting. Differences in FEN1 mRNA expression in AML patients achieving different therapeutic effects were compared among AML patients whose data with evaluable efficacy. AML patients were divided into high FEN1 expression group (≥ critical value) and low FEN1 expression group (< critical value), taking the median relative expression level of FEN1 mRNA as the critical value. The correlation of FEN1 expression level with clinicopathologic features, laboratory indicators, cellular and molecular genetic changes in AML patients at initial diagnosis was analyzed.Results:The median relative expression of FEN1 mRNA in newly treated AML patients was higher than that in healthy controls [0.696 (0.025-3.661) vs. 0.246 (0.013-1.237), Z = 1.75, P = 0.041]. Western blotting showed that the expression level of FEN1 protein in AML patients was higher than that in healthy controls. The relative expression of FEN1 mRNA in 15 recurrent AML patients was higher than that in 19 patients patients achieving complete remission (CR) [1.153 (0.047-4.172) vs. 0.259 (0.023-1.148), Z = 2.71, P = 0.009]. The proportion of patients with French-American-British(FAB) type M 5, fever at initial diagnosis and lymph node enlargement in FEN1 high expression group (32 cases) was higher than that in FEN1 low expression group (25 cases) (all P < 0.05). There were no significant differences in the proportion of gender, age, fatigue, pale skin mucosa and large liver and spleen of patients between the two groups (all P > 0.05). At initial diagnosis, the white blood cell count, lactate dehydrogenase, C-reactive protein and bone marrow primitive cell proportion in FEN1 high expression group were higher than those in FEN1 low expression group (all P < 0.05), and the hemoglobin and platelet count in FEN1 high expression group were lower than those in FEN1 low expression group (all P < 0.05). There were no significant differences in procalcitonin level, the proportion of chromosome karyotype, cytogenetic prognosis grade and patients with or without gene mutation between the two groups (all P > 0.05). Conclusions:FEN1 expression is up-regulated in AML patients and further increased in relapsed patients. FEN1 expression in AML patients is associated with adverse clinicopathological features and poor detection results of laboratory indicators, which may become indicators for disease monitoring in AML patients.

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.

Research progress of fluoride on non-skeletal injury in multiple systems of the body / 中华地方病学杂志

Fluorine is an essential trace element for human body. Appropriate amount of fluorine can have a certain positive impact on the body, but long-term intake of excessive fluorine will cause damage to the body, which is an important cause of endemic fluorosis. The main feature of fluorosis is skeletal injury to the bones and teeth of the body. In recent years, the research of non-skeletal injury has been increasingly paid attention by research scholars. This paper summarized the non-skeletal injury of fluorine to urinary, digestive, nervous, circulatory, reproductive, immune systems and other systems, providing a reference for fluorosis-related studies other than skeletal injury.

Construction of clinical medical examination question bank based on virtual reality technology / 中华医学教育探索杂志

Using novel virtual reality (VR) technology to carry out the construction of clinical medical examination question bank, while deepening the reform of clinical medical course examination, it continues to innovate the medical professional evaluation system and improve the flexibility, diversity and scientificity of clinical medicine in teaching and assessment. It is of great and far-reaching significance to improve the teaching level and the quality of medical education in medical colleges and universities. This paper analyzes and discusses the necessity and feasibility of building a clinical medical examination question bank, and the advantages and prospects of integrating VR technology to carry out the construction of clinical medical examination question bank. At the same time, the exploration and practice of the examination question bank construction based on VR technology disscussed in detail would provide innovative thinking and reference for the clinical medical teaching and evaluation, medical personnel training and other aspects in China.

Effects and mechanism of digoxin on atrium electrical remodeling and susceptibility of atrial fibrillation in aged rabbits / 中华心血管病杂志

Objective@#To investigate the effects and mechanism of digoxin on atrium electrical remodeling and susceptibility of atrial fibrillation (AF) in aged rabbits.@*Methods@#Twenty aged male New Zealand rabbits were divided into aged group and aged plus digoxin group (n=10 each). Electrical parameters including heart rate (HR), RR and QT interval, ST segment and P wave dispersion from normal Ⅱ electrocardiogram, and the maximum upstroke velocity (Maxdv/dt), plateau potential (plateau P), action potential duration of 10%, 20% and 90% (APD10, APD20, APD90) from recording of monophasic action potential (MAP), as well as atrial effective refractory period (AERP200) and dispersion (dERP200) with 200 ms of basic cycle length (BCL), and frequency self adaptation of AERP with 300 ms and 150 ms of BCLs (fERP) were recorded and compared between the 2 groups. BCLs and inducibility of AF post programmed electrical stimulation and Burst-pacing in left atrium tissue of rabbits in vivo were also analyzed. The L-type calcium current (ICa-L) in 2 groups were recorded via whole-cell patch clamp technique, and the fluorescence intensity of intracellular free Ca2+ was detected with Flup-3/AM loading by the laser scanning confocal microscope in enzymatically dissociated single rabbit atrial myocytes.@*Results@#Compared with aged group, the heart rate was faster, RR and QT interval were obvious shorter, ST segment was raised and P wave dispersion was significantly increased in aged plus digoxin group (all P<0.05). Moreover, compared with aged group, the Maxdv/dt and plateau P were obviously increased, APD10 and APD20 were significantly prolongated, and APD90 was significantly shorter in aged plus digoxin group (all P<0.01). Otherwise, the fERP was markedly increased (0.81±0.15 vs. 0.67±0.05), and the induced rate of AF was obviously higher in aged plus digoxin group than in aged group (6/8 vs. 4/9) (all P<0.01). With voltage clamp model, digoxin significantly increased ICa-L of atrial myocytes of aged rabbits, When command potential was 10 mV, the current densities of ICa-L were significantly higher in digoxin group than that in aged group ((15.45±2.38) pA/pF vs. (7.03±1.69) pA/pF, P<0.01). Otherwise, the I-V curve of ICa-L was downward shifted of all I-V curves in digoxin perfused aged atrial cells of rabbits. Moreover, the fluorescence intensities of intracellular free Ca2+ was significantly higher in aged plus digoxin group than in aged group ((1 748±173) μmol/L vs. (478.13±87.63) μmol/L, P<0.01).@*Conclusion@#Digoxin could aggravate the atrial electrical remodeling in atrium of aged rabbits, facilitate susceptibility of atrial fibrillation in aged rabbit, increased current density of ICa-L and concentration of intracellular free Ca2+, followed Ca2+ overload and oscillations might be part of the underlying mechanisms.

Clinical Observation ofTongYuanNeedling Method plus Letrozol and HCG for Refractory Sterility Caused by Polycystic Ovary Syndrome / 上海针灸杂志

Objective To observe the clinical efficacy ofTong Yuan(short forTong Du Tiao Shen,Yin Qi Gui Yuan, referring to unblocking the Governor Vessel, regulating mind, and guiding qi back to the origin) needling method plus Letrozol and human chorionic gonadotropin (HCG) in treating refractory sterility caused by polycystic ovary syndrome (PCOS).Method Sixty clomiphene citrate-resistant PCOS-induced sterility patients were recruited and randomized into two groups. The treatment group was intervened byTong Yuanneedling plus Letrozol and HCG, while the control group was intervened only by Letrozol and HCG, both for 3 months. Prior to and after the treatment,the levels of estradiol (E2), follicle stimulating hormone (FSH), luteinizing hormone (LH), and testosterone (T) were detected; the endometrium, ovulation and pregnancy rates were also observed.Result After 3 menstrual cycles, the level of FSH increasedsignificantly (P<0.05), LH decreased significantly (P<0.05), LH/FSH declined significantly (P<0.05), and the level of T also dropped significantly (P<0.05) in both treatment group and control group; the level of E2also dropped significantly after the treatment in both groups (P<0.05), and the decrease in the treatment group was more significantly than that in the control group (P<0.05). The ovulation and pregnancy rates in the treatment group were significantly higher than those in the control group (P<0.05).Conclusion Tong Yuanneedling plus Letrozol and HCG can significantly down-regulate the levels of LH, E2and T, up-regulate the level of FSH, effectively improve the internal reproductive environment of refractory PCOS patients, and enhance the ovulation and pregnancy rates.

Effects of cardiac remodeling on atrial arrhythmogenesis in aged rabbit left atrium / 中国病理生理杂志

AIM: To investigate the effects of myocardial remodeling of aged left atrium (LA) on atrial ar-rhythmogenesis in rabbits .METHODS:The male New Zealand rabbits were divided into young LA and aged LA groups . By observing the changes of monophasic action potential ( MAP) and burst-pacing in LA of the rabbits in vivo, the main cardioelectrophysiological parameters such as resting membrane potential ( RMP) , action potential amplitude ( APA) , ma-ximum rise veloctiy of action potential (dv/dtmax), plateau potential and action potential duration at 30%, 50%and 90%( APD30 , APD50 and APD90 ) , as well as the inducibility and duration of atrial arrhythmias were recorded .L-type calcium current (ICa,L) was analyzed via whole-cell patch-clamp technique in enzymatically dissociated single rabbit LA myocytes . The myocardial collagen content was quantified after Masson staining , and the ultrastructure of the LA cells was observed under scanning electron microscope .The expression of Cav 1.2 in LA tissue of the 2 groups was detected by Western blot . RESULTS:Compared with young LA group , dv/dtmax and plateau potential were significantly decreased , APD30 and APD50 were shortened, and APD90 was notably prolongated in aged LA group (P<0.01).The inducibility or duration of atrial arrhythmias was severely increased or prolongated in aged LA group (P<0.01).With voltage clamp model, the den-sity of ICa,L in aged LA group was significantly decreased , and current-voltage curve was notably moved upward compared with young LA group.When the clamp potential was +20 mV, the density of ICa,L was notably modified from (11.72 ± 1.39) pA/pF in young LA group to (6.08 ±0.98) pA/pF in aged LA group ( P<0.01).Compared with young LA group, the protein level of collagen was significantly increased (P<0.01), and the arrange of atrial myocytes was irregular in LA of rabbits in aged LA group .The atrial myocytes of the LA wall in aged LA group exhibited abnormal ultrastructural alterations , such as karyopyknosis , irregular and swelling mitochondria with the presence of vacuoles , and mild and severe sarcomere degeneration .Compared with those in LA tissues of young rabbits , the expression levels of Cav 1.2 in the LA tis-sues of aged rabbits were severely reduced (P<0.01), and had a significant positive correlation with the reduction of ICa,L (r=0.83, P<0.01).CONCLUSION:The pathophysiological characteristics of aged LA are significantly altered , and might contribute to vulnerability and susceptibility of occurrence of atrial fibillation in aged rabbits .The mechanisms might completely attribute to the notable reduction of ICa,L , abnormal alterations of ultrastructures and obvious decrease in the ex-pression of Cav1.2 in the aged LA of aged rabbits .

The influence of U0126 of ERK signaling pathway inhibitor on early brain inj ury and neurons autophagy in SAH rats / 西安交通大学学报(医学版)

ABSTRACT:Objective To explore role of U0126,the specific inhibitor of ERK signaling pathway,in early brain injury (EBI)and the autophagy of nerve cells in hippocampus area in subarachnoid hemorrhage (SAH). Methods A total of 48 male adult SD rats were randomly divided into control group,SAH group,DMSO+SAH group,and U0126+SAH group,with 12 in each.We established SAH rat model by the puncture of internal carotid artery.The same amount of saline water,DMSO and U0126 solution of 0.5 mL per rat was injected respectively into the rats of different groups 30 min before modeling.The rats were killed at 24 h.To measure brain water content by Wet and dry method after 24 h,the morphological changes of hippocampus CA1 neural cells were observed by microscopy;the expression levels of ERK,Beclin-1 and LC3 were detected by using immunohistochemical method. Results Compared with that in sham group,brain water content increased obviously in SAH model group.The density of surviving neurons in SAH group was significantly lower than that in control group (P<0 .0 5 ).ERK signaling pathway was activated obviously,the expressions of Beclin 1 and LC3-Ⅱ were significantly higher than those in control group (P<0.05).Compared with SAH model group,in U0126 group brain water content increased obviously.Compared with those in SAH group,the density of surviving neurons was significantly lower (P<0.05), ERK signaling pathway was suppressed,the expressions of Beclin-1 and LC3-Ⅱ were significantly lower (P<0.05). Conclusion The U0126,the ERK signaling pathway inhibitor,can inhibit neuron autophagy and increase EBR of SAH.

Effects of miR-7-5p on TK6 cell damage induced by hydroquinone / 中国职业医学

OBJECTIVE: To investigate the mechanism of miR-7-5p in TK6 cell damage induced by hydroquinone( HQ) by constructing stable miR-7-5p over-expressing human lymphoblastoid TK6 cell line using lentivirus. METHODS: i) The miR-7-5p over-expression lentivirus vectors were constructed,and then infected to TK6 cells. The miR-7-5p overexpression stable TK6 cell line( TK6-miR-7-5p cells) and negative control cells( TK6-NC cells) were selected with puromycin. The infection efficiency was confirmed by real-time fluorescent quantitative polymerase chain reaction assay.ii) TK6,TK6-NC and TK6-miR-7-5p cells were treated with HQ at final concentrations of 0 and 40 μmol/L for 48 hours.Cell viability was determined by CCK-8 assay. The early apoptosis rate of cells was detected by flow cytometry. The relative expression of poly( ADP-ribose) polymerase-1( PARP-1) and breast cancer susceptibility gene 1( BRCA1) proteins in 3 kinds of cells treated with HQ at the final concentration of 40 μmol/L was detected by Western blotting. RESULTS: i) The TK6 cell line with stable expression of miR-7-5p were successfully screened. Compared with normal TK6 cells,the relative expression of miR-7-5p in TK6-miR-7-5p cells increased by about 17 times( P < 0. 01) with no significant changes in cell morphology. ii) After treatment with 40 μmol/L HQ,the survival rate of TK6-miR-7-5p cells decreased compared with normal TK6 cells and TK6-NC cells( P < 0. 01),early apoptosis rate increased( P < 0. 01),the relative expression of PARP-1 and BRCA1 protein was decreased( P < 0. 05). CONCLUSION: MiR-7-5p may lead to the increase of early apoptosis in TK6 cells induced by HQ through inhibiting the DNA damage repair capacity related to PARP-1 and BRCA1.

Application of a modified culturing method for lymphocytes in cytogenetic research / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To establish a modified method for microculturing whole human blood for cytogenetic analysis.</p><p><b>METHODS</b>A novel tube rack was designed to overcome the drawbacks of directly culturing the cells within centrifuge tubes. The fractions of human plasma, human serum and two commercial fetal bovine sera were analyzed with 15% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The influence of adding 0%, 5%, 10%, 15%, 20%, 25% and 30% autologous plasma to the culture on lymphocyte transformation rate and mitotic index (MI) was examined.</p><p><b>RESULTS</b>The SDS-PAGE analysis showed a significant difference between commercial fetal bovine sera, and that the components of human plasma were similar to those of fetal bovine serum. The value of MI in lymphocyte was evidently increased along with addition of autologous plasma. However, this has exerted no significant effect on the transformation rate. With the addition of 10% autologous plasma, the MI value has become much higher than the conventional method.</p><p><b>CONCLUSION</b>A modified method was established by application of a novel tube inclined rack and optimization of whole blood inoculation. This method is easier and cheaper, and is suitable for application in clinical practice.</p>

Surface display of lead-binding protein PbrR on Escherichia coli and preliminary study of intestinal ;colonization by the recombinant bacteria / 中华微生物学和免疫学杂志

Objective To construct a recombinant Escherichia coli ( E. coli) with surface-dis-played lead specific binding protein PbrR and to further study intestinal colonization by the recombinant bac-teria in mice and gastrointestinal tolerance of the bacterial surface-displayed PbrR. Methods Chimeric pro-tein Lpp-OmpA coding sequence was chemically synthesized and inserted into the expression vector pET-21a to construct the outer membrane display vector pLOA. PbrR coding sequence was also obtained by chemical-ly synthesis and inserted into pLOA to generate the outer membrane display plasmid pLOA-pbrr. E. coli BL21 (DE3)pLysS was transformed with pLOA-pbrr and induced by IPTG. The expressed recombinant proteins were analyzed by 15% SDS-PAGE and Western blot assay. Lead adsorption capacity of the cell surface-dis-played PbrR in the simulated intestinal juice and tolerance of the recombinant E. coli to simulated gastric juice were analyzed, respectively. KM mice were orally given the induced recombinant bacteria by gastric lavage for 7 consecutive days and then were continually fed until day 30. The contents of recombinant bacte-ria in stool samples were detected by dilution plate method on day 7, 15 and 30. The recombinant protein with His tag was detected by immunoblotting on day 7 and 15. Results Based on Lpp-OmpA, the PbrR outer membrane display vector was successfully constructed. The recombinant fusion protein Lpp-OmpA-PbrR-His tag was highly expressed in E. coli. The recombinant E. coli strains displaying PbrR on their outer membrane accumulated a significant level of Pb2+ in simulated intestinal juice. Moreover, those strains showed a tolerance to gastric acid in vitro and could colonize in the intestinal tracts of mice via oral infection. The surface-displayed recombinant fusion protein showed a better tolerance to the environment of digestive tract. Conclusion The recombinant E. coli strain displaying PbrR on its surface showed a stronger capabili-ty of lead accumulation from simulated intestinal environment and could colonize in the intestinal tracts of mice. The surface-displayed recombinant PbrR also showed a good tolerance to digestive juice. This study paved the way for further researches on the selective elimination of lead by biosorption based on animal mod-els.

Simultaneous Determination of the Costunolide and Dehydrocostus Lactone in Haoweilai Soft Capsule by HPLC / 中国药房

OBJECTIVE:To establish a method for the simultaneous determination of costunolide and dehydrocostus lactone in Haoweilai soft capsule. METHODS:HPLC was performed on the column of Dikma C18 with mobile phase of acetonitrile- water (V/V,55∶45) at a flow rate of 1.0 ml/min,detection wavelength was 225 nm,column temperature was 25 ℃,and the injection volume was 10 μl. RESULTS:The linear range were 24.00-108.00 μg/ml(r=0.999 7) for costunolide and 20.88-93.98 μg/ml for dehydrocostus lactone (r=0.999 8);RSDs of precision,stability and reproducibility tests were lower than 1%;recoveries were 98.71%-100.00%(RSD=0.25%,n=6)and 96.88%-99.18%(RSD=0.40%,n=6). CONCLUSIONS:The method is simple with good stability and reproducibility,and can be used for the simultaneous determination of costunolide and dehydrocostus lactone in Haoweilai soft capsule.

Effect of Extracellular Regulated Protein Kinases Signaling Pathway on Early Brain Injury and Neurons Autoph-agy in Rats with Subarachnoid Hemorrhage / 中国康复理论与实践

Objective To explore the effect of extracellular regulated protein kinases (ERK) signaling pathway on early brain injury and autophagy of nerve cell in hippocampus area in rats with subarachnoid hemorrhage (SAH). Methods Forty-eight adult male Sprague-Daw-ley rats were randomly divided into sham group, SAH group, SAH+dimethyl sulfoxide (DMSO) group and SAH+U0126 group, with 12 rats in each group. The SAH model was established with puncture of internal carotid artery. The SAH+U0126 group was injected with U0126 0.05 mg/kg;the sham group and SAH group were injected with normal saline, and the SAH+DMSO group was injected with DMSO 30 min-utes before modeling. They were sacrificed 24 hours after modeling. The brain water content was measured with wet and dry method. The morphology changes of neural cells in hippocampus CA1 were observed by HE staining. The expression of phosphorylation ERK (p-ERK), Beclin-1 and LC3-Ⅱwere detected with immunohistochemical method and Western blotting. Results Compared with the sham group, the brain water content increased (P0.05). Conclusion The activation of ERK signaling pathway may alleviate early brain injury after SAH by regulation of autophagy.

Clinical Observations on Heat-sensitive Point Dazhui (GV14) Moxibustion for the Treatment of Cervical Spondylotic Radiculopathy / 上海针灸杂志

Objective To investigate the clinical efficacy of heat-sensitive point Dazhui(GV14) moxibustion in treating cervical spondylotic radiculopathy. Method Ninety-six patients with cervical spondylotic radiculopathy were randomly allocated to a heat-sensitive point Dazhui moxibustion (observation) group of 30 cases, a non-heat-sensitive point Dazhui suspended moxibustion (control) group of 33 cases and a medication group of 33 cases. The observation group received heat-sensitive point Dazhui moxibustion; the control group, non-heat-sensitive point Dazhui suspended moxibustion; the medication group, oral administration of Jingtong granules. The clinical symptoms were assessed and serum hypersensitive C-reactive protein (HsCRP) and interleukin-8 (IL-8) were measured in the three groups before and after 15 days of treatment. Result All the three treatments had a certain effect on cervical spondylotic radiculopathy. The therapeutic effect was best in the observation group (P0.05). Conclusion The therapeutic effect of heat-sensitive point Dazhui moxibustion is better than those of non-heat-sensitive point Dazhui suspended moxibustion and oral Jingtong granules. It is a more ideal way to treat cervical spondylotic radiculopathy. The mechanism of its action may be related to post-treatment decrease in inflammatory factors in the lesion.

Free fibula flap and computed tomographic angiography in the functional reconstruction of oral and maxillofacial hard and soft tissue defects / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#Assessed the feasibility of application of free fibular flap and clinical significance of pre-operational contrast enhanced CT angiography in functional reconstruction of oral and maxillofacial hard and soft tissue defects.@*METHOD@#Eight cases with mandibular and soft tissue defects received a free fibula flap using arteriovenous anastomosis anastomosis method. The relationship between fibula flap design, size, repair parts and survival were analyzed. Preoperative enhanced CT angiography (CTA) examination was conducted to detect any abnormal blood vessels in fibula flap valve area.@*RESULT@#Peroneal artery and posterior tibial artery variation was identified in one case of gums cancer, who used other muscle flap. Free fibula flap in the other 7 cases survived, which carried a skin island with an area ranging from 6 cm x 2 cm to 10.0 cm x 3.5 cm. Postoperative facial appearance, functionality, dental occlusion and voice function was normal. Lower limb function returned to normal from 2 weeks to 4 months after surgery.@*CONCLUSION@#CTA examination of the free vascularized fibula flap in the preoperative evaluation of the donor site is a valuable tool. Free fibula flap of bone, periosteum and bone marrow has a dual blood supply and are highly resistant to infection after transplantation. Personalized shaping osteotomy allowed for accurate recovery of mandibular alveolar patterns. Furthermore, the height and width of the fibula and the thickness of cortical bone is suitable for dental implants. Free fibula flap skin island can also be used to monitor the post-operative blood supply and is an ideal technique for mandible and soft tissue defects reconstruction as well as functional restoration.

Correlation between caspase regulatory gene expression and facial nerve injury in a facial nerve injury model / 中国组织工程研究

BACKGROUND:Caspase plays a crucial role in the cellapoptosis, but the influence of different facial nerve injury on caspase 1, caspase 8, cyto-c protein expression and their correlation stil remain unclear. OBJECTIVE:To construct facial nerve crush or distal transection injury models, observe the morphological changes of facial motoneurons, investigate death gene caspase 3, caspase 8, cyto-c expression, and analyze their correlation. METHODS:Facial nerve crush or distal transection injury model was established in the right facial nerve of rats, while the left facial nerve served as normal controls. We observed the morphology and the death of facial motoneurons with toluidine blue staining and transmission electron microscope. Expressions of caspase 3, caspase 8 and cyto-c proteins were studied by immunohistochemistry analysis fol owing facial nerve injury. RESULTS AND CONCLUSION:Both facial nerve distal transection and crush injury resulted in the death of facial motoneurons, and the death pattern was mainly apoptosis. Caspase 3, caspase 8 and cyto-c protein expressions were observed in the subnucleus of normal rat facial nucleus. cells of the distal transection group were stained more intensely than that of crush group. Expressions of these proteins began to increase at 3 days after the injuries. Caspase 3 and caspase 8 protein expression peaked at 14 days, whereas cyto-c protein expression peaked at 7 days after the injuries. Expressions of caspase 3, caspase 8 and cyto-c proteins were correlated with facial nerve injury type and injury time. Expressions of caspase 8 and cyto-c protein were correlated with expression of caspase 3 protein. The findings indicate that, caspase 8 and cyto-c contribute to activate caspase 3, and caspase cascade reaction plays an important role in the apoptosis of facial motoneurons.