The Ⅳ stage esophageal cancer refers to exist regional lymph nodes and distant organs in patients with metastases. Palliative chemotherapy and radiotherapy is the primary treatment. With the deepening of the clinical study,radiotherapy and chemotherapy are no longer single treatment,chemotherapy combined molecular targeted drugs,chemo-radiotherapy and some new treatment methods have been gradually recognized, and especially molecular targeted drugs began to establish its position in the treatment of tumor. Application of various methods individual or combined treatment can effectively prolong survival and improve the quality of life.

To isolate and culture the purified monoclonal neural stem cells from the cerebral cortex of new born mice, new-born mice cerebral cortex was isolated and dissociated to single-cell suspension by mechanical trituration. The dissociated single cells were cultured in serum-free medium. After the formation of neurospheres, single-cell clone culture was performed by limiting dilution and the proliferated single-cell clones were harvested for subculture. Immunocytochemistry was used to detect the specific marker of neuroepithelial stem cells (Nestin) of the primary and monoclonal neurospheres. In the differentiated cells we detected the specific antigen of NF-200 and GFAP. Our results showed that the primary neurospheres expressed Nestin antigen positively. By limiting dilution, we cultured the cell lines from single-cell clone and the monoclonal neurospheres expressed Nestin and had capabilities of self-renewal, proliferation and the potentiality of differentiation into neurons and glial cells. It is concluded that monoclonal neural stem cells which have the ability of proliferation and multi-directional differentiation can be isolated and cultured from the cerebral cortex of new-born mice by limiting dilution.

BACKGROUND: Growth-associated protein-43, a kind of protein relatedto axonal growth, plays a key role in promoting neural development, axonalregeneration, synaptic growth and structural and functional reconstructionand so on. In the study, we find that cyclovirobuxine D can protect braininjury in rats with experimental cerebral ischemia reperfusion.OBJECTIVE: To observe the effect of cyclovirobuxine D on the expression of growth-associated protein-43 mRNA in hypertensive rats with cerebral ischemia reperfusion.DESIGN: A randomized controlled animal experiment.SETTING: Foshan Hospital of Traditional Chinese Medicine of Guangdong Province; Central Laboratory of Guangdong Hospital of Traditional Chinese Medicine.MATERIALS: Cyclovirobuxine D is alkaloid monomer extracted from Chinese herb buxine. Cyclovirobuxine D powder with national protected traditional medicine number of ZYB20796057 was provided by Nanjing Xiaoyingyao Pharmaceutical Factory. Totally 120 two-to-three-month-old healthy male SD rats, of either gender, with body mass of 90 to 120 g, were used in this experiment.METHODS: This experiment was carried out in Foshan Hospital of Tra ditional Chinese Medicine of Guangdong Province and at the Central Laboratory of Guangdong Hospital of Traditional Chinese Medicine between June 2005 and March 2006. ①Stroke-prone-renovascular-hypertensive-rats models (RHRSP)were created by bilaterally narrowing the renal artery with silk loop clips. Totally 120 rats were randomly divided into blank group (n=20,renovascular hypertensive rats were given no treatments), sham operation group (n=20, rats were given only surgical trauma), model group (n=40, rats were given treatment of cerebral ischemia reperfusion) and cyclovirobuxine D-treated group (n=40, rats were given cyclovirobuxine D).② Unilateral occlusion of the middle cerebral artery ischemia reperfusion models were made with suture-occluded method. 6.48 mg/kg cyclovirobuxine D diluted by 1.5 mL normal saline was intraperitoneally injected into the rats of cyclovirobuxine D-treated group, twice per day; Normal saline was isochronously intraperitoneally injected into the rats of each subgroup of control group, 2 mL once, the method was the same as that of the cyclovirobuxine D-treated group; interval of injection time was 7 hours in each group. Rats in each group were executed on days 1, 7, 14 and 30after ischemia reperfusion. ③ Brain slice was prepared. The expression of growth-associated protein-43 mRNA of rats in each group was detected with in situ hybridization.MAIN OUTCOME MEASURES: ① The expression of growth-associated protein-43 mRNA around ischemia are following ischemia reperfusion for 2 hours. ②The expression of growth-associated protein-43 mRNA in hippocampus following ischemia reperfusion for 2 hours RESULTS: All the 120 rats entered the stage of result analysis. ① Immune in situ hybridization of growth-associated protein-43 mRNA: In situhybridization showed that expression of growth-associated protein-43 mRNA and expression of growth-associated protein-43 mRNA could be detected respectively in the hippocampus and marginal area of hematoma after ischemia and reperfusion.②The expression of growth-associated protein-43RNA around haematoma following cerebral ischemia reperfusion group as not found in blank group and sham-operation group; The expression of growth-associated protein-43 mRNA was found in the marginal zones around haematoma of rats in the model group on the 1st day following ischemia reperfusion and it was significantly increased on the 7th day, gradually reduced on the 14th day and still expressed on the 30th day but less,with significant difference at each time point (P ＜ 0.01); Expression of growth-associated protein-43 mRNA around haematoma at each time point was more in the cyclovirobuxine D-treated group than in the model group ,with significant difference (P ＜ 0.05). ③There was no significant difference of the expression of growth-associated protein-43 mRNA in hippocampus of rats following cerebral ischemia reperfusion between blank group and sham-operation group; Expression of growth-associated protein-43 mRNA was found in hippocampus of rats in the model group on the 1st day after modeling, and it reached the peak on the 7th day, gradually decreased on the 14th day and significantly decreased on the 30th day, but significantly more than that of sham-operation group; The expression of growth-associated protein-43 mRNA in hippocampus was significantly more at each time point in cyclovirobuxine D-treated group than in model group, with significant difference (P ＜ 0.01).CONCLUSION: Cyclovirobuxine D up-regulates the expression of growthassociated protein-43 mRNA after reperfusion and promotes axonal regeneration of rats with experimental cerebral ischemia.

BACKGROUND: Deletion of glial cell-derived neurotrophic factor and endothelin receptor type B gene will induce abnormal development of enteric nervous system. Neural stem cell transplantation can repair nervous system from anatomy and function,and be considered as a vector of gene transfection.OBJECTIVE: To transfect recombinant adenovirus carrying glial cell-derived neurotrophic factor and endothelin receptor type B gene into mouse neural stem cells, and to observe expression of target gene.DESIGN: A cell-gene study.MATERIALS: New-born Kunming mice were provided by the Animal Center of Tongji Medical College, Huazhong University of Science and Technology, China. jetPEI reagent was purchased from PolyPlus Co, France. The pAdTrack-CMV-GE with green fluorescent protein (GFP) was gifted by Doctor Sun Nianfeng and Zhang Jinghui in our laboratory.METHODS: Neonatal mouse brain tissues were sterilely obtained to prepare monoplast suspension. Adenovirus expressing glial cell-derived neurotrophic factor and endothelin receptor type B gene with GFP was dissolved in NaCI to prepare JetPEI/DNA complex. Subcultured neural stem cells in DMEM/F12 were regulated to 5×10~8/L, and 400 μL cell suspension and 100 μL JetPEI/DNA complex were seeded on a 24-well plate at 37 ℃ in 5% CO_2 incubator. Neural stem cells were harvested at 24, 48 and 72 hours following transfection.MAIN OUTCOME MEASURES: The efficiency of transfection was detected using fluorescence microscope and flow cytometry.Target gene expression in neural stem cells was determined using RT-PCR.RESULTS: Bright green fluorescence of the transfected cells could be observed under fluorescence microscope after 24 hours of transfection. The positive rate of GFP was 15.36%, 24.67%, 25.73% at 24, 48 and 72 hours following transfection respectively.Neural stem cells expressed glial cell-derived neurotrophic factor and endothelin receptor type B gene at various time points.Strap brightness was low at 24 hours, and exogenous gene expression was great at 72 hours.CONCLUSION: The target genes were successfully transfected into neural stem cells by using jetPEI reagent. Moreover, glial cell-derived neurotrophic factor and endothelin receptor type B gene effectively transcribed and expressed in target cells.

Objective To investigate the diagnostic value of Multislice CT scanning in patients with acute thoracic trauma. Methods151 patients with chest wound admitted after Multislice CT scanning were reviewed and analyzed. ResultsThis group of 151 patients with chest wound after X-ray and CT inspection were as follows:lungs were damaged in 139 cases including 52 cases with traumatic wet lung,94 cases with lung rip,28 cases with lacerated wound and 7 cases with tracheal bronchial tube damage.Damages outside the lungs were found in 108 patients including 83 patients with trauma in pleural membrane(hemothorax 27 cases,pneumothorax 32 cases and blood pneumothorax 24 cases),mediastinum damage in 9 cases(vertical mediastinal pneumatosis 6 cases,hematocele 4 cases and traumatic disphragmatic hernia 2 cases),thoracic wall damage in 76 cases(rib bone fracture 59 cases,breast bone fracture 18 cases and costal cartilages damage 5 cases).Clavicle and scapula fracture in 31 cases. ConclusionMultislice CT scanning was principal method for the diagnosis of chest wound,it had the advantages of scanning range,quick scanning and characteristics of high sensitivity,and it would certainly play a vital role in first aid process in emergency medical treatment of wound.

Objective To explore the relationship between the overlap volume of organ at risk (OAR) and target (Voverlap) and the mean dose to OAR (DmOAR) in intensity-modulated radiotherapy (IMRT).Methods Fifty randomly selected patients undergoing IMRT for nasopharyngeal carcinoma (NPC) and an equal number of patients undergoing radical IMRT for cervical cancer in our hospital were enrolled as subjects.The relationship between Voverlap and DmoAR in IMRT was analyzed.The Matlab software was used to generate function to fit the relationship between Voverlap and DmOAR for different OARs.Results The Voverlap varied among patients with NPC or cervical cancer.The ratio of Voverlap to the volume of OAR (VOAR) was positively correlated with the ratio of DmOAR to the prescribed dose (Dp) (all P=0.01).A function was generated to fit the correlation between Voverlap/VOAR ratio and DmOAR/Dp ratio.Conclusions In IMRT,patients have different Voverlap/VoAR ratios due to varicd target volume,disease stages,OAR filling status,and OAR volume.As criteria for plan verification,a specific DmOAR value for each OAR should be calculated before radiotherapy planning based on the corresponding correlation function and the Voverlap/VoAR ratio with a fixed dose prescription.It can be used to reduce the subjective influence on the optimization of radiotherapy planning.

Objective : To investigate the epidemiological and clinical characteristics of cases with pertussis in Yiwu City, Zhejiang Province, so as to provide insights into pertussis diagnosis and control. Methods: A total of 186 patients with definitive diagnosis of pertussis in medical institutions in Yiwu City from November 1, 2020 to August 31, 2022 were recruited, and subjects' demographic, clinical symptoms and history of pertussis vaccination were collected using questionnaire surveys. The temporal, population, and spatial distribution and clinical symptoms of pertussis were analyzed using a descriptive epidemiological method, and the clinical characteristics of pertussis patients with different doses of pertussis vaccination were compared. Results: Pertussis was found to predominantly occur during the period between July and November (101 cases, 54.30%), and the three highest-incidence regions included Jiangdong Street, Beiyuan Street and Choujiang Street (87 cases, 46.77%). The 186 pertussis cases included 105 males (56.45%) and 81 females (43.55%), and included 144 cases with age of onset under 7 year (77.42%). Preschool and diaspora children were predominant among all pertussis cases, and the main clinical symptoms included spasmodic cough (97 cases, 52.15%), post-tussive vomiting (82 cases, 44.09%) and aggravated cough at night (77 cases, 41.40%). Routine blood tests measured 119 cases with abnormal white blood cell counts (63.98%), 137 cases with abnormal lymphocyte counts (73.66%), 39 cases with abnormal neutrophil counts (20.97%), 21 cases with abnormal platelet counts (11.29%) and 111 cases with abnormal hemoglobin concentrations (59.68%). There were 55 cases that were unvaccinated (29.57%), 23 cases that were not fully vaccinated (12.37%), and 108 cases that were fully vaccinated (58.06%). There were significant differences among pertussis cases with different doses of vaccination in terms of age, incidence of post-tussive vomiting, percentage of abnormal platelet counts and percentage of hemoglobin concentrations (all P<0.05). Conclusions The majority of pertussis cases are preschool and diaspora children in Yiwu City from November 1, 2020 to August 31, 2022, and the clinical symptoms mainly include spasmodic cough, post-tussive vomiting and aggravated cough at night, with atypical symptoms. The capability for differential diagnosis of pertussis is required to be improved in medical institutions.

Objective To study the ages at natural menopause of the women in Jilin Province, and to illustrate its influencing factors among the women in Jilin Province.Methods Through multistage stratified cluster random sampling method,23 050 people aged from 18 to 79 years were drew from nine states(a total of 32 areas)of Jilin province.The data of these residents were collected with the questionnaire and physical examinations by face-to-face interview.The number of selected female sample was 11 098. Finally, 4 881 postmenopausal women were selected.Complex weighted computation was used to estimate the ages at natural menopause.One-way ANOVA was used to compare the ages at natural menopause of the women with different birth years. Multiple linear regression analysis were used to examine the influencing factors of the ages at natural menopause. Results The mean and median ages at natural menopause were (49.11±4.19)years and 50.00 years,respectively.There were 4 881 cases of postmenopausal women,among them the women with age at natural menopause<40 years,40 year≤age at natural menopause≤45 years,46 years≤ age at natural menopause≤53 years,age at natural menopause≥54 years and age at natural menopause missing accounted for 2.27%(111 cases),13.17%(643 cases),71.97%(3 513 cases),11.74% (573 cases),and 0.85%(41 cases),respectively.Converted to birth years by age,70-79 years old was 1933-1942 birth years,60-69 years old was 1943-1952 birth years and 57-59 years old was 1953-1955 birth years.The age at natural menopause in Jilin province was statistically significant among the women with different birth years(F=21.178,P<0.001).By SNK-q test among three different birth year groups, the age at natural menopause was different between any two groups among three different birth year groups and the ages at natural menopause of 1953-1955 birth year group,1943-1952 birth year group and 1943-1952 birth year group were 50.38 years,49.51 years and 48.81 years.The age at natural menopause in urban of Jilin province was statistically significant among the women with different birth years(F=16.633,P<0.001).By SNK-q test among three different birth year groups,the age at natural menopause was different between any two groups among three different birth year groups and the ages at natural menopause of 1953-1955 birth year group,1943-1952 birth year group and 1943-1952 birth year group were 50.77 years,49.73 years,and 48.85 years,respectively.The age at natural menopause in rural of Jilin province was statistically significant among the women with different birth years(F=7.400,P=0.001 ). By SNK-q test among three different birth year groups, the age at natural menopause was different between 1953-1955 birth year group and the other two groups and the ages at natural menopause of 1953-1955 birth year group,1943-1952 birth year group and 1943-1952 birth year group were 50.09 years,49.33 years,and 48.74 years,respectively.The multiple linear regression results indicated that BMI and exercise were positively correlated with the age at natural menopause,but smoking and mental health evaluation were negatively.Consumption frequency of vegetables,fruits,bean products,and meat was no correlated with the age at natural menopause.Conclusion The differences of the ages at natural menopause between the women with different birth years are statistically significant in Jilin Province;BMI, smoking, exercise,and mental health are the influencing factors of the age at natural menopause.

OBJECTIVE To investigate the effect of doxazosin(DOX) and metoprolol( MET) on vascular remodeling in rats with abdominal aorta coarctation (AAC). METHODS An animal model was established by AAC. Two weeks later, the rats were treated with DOX (10 mg.kg-1 per day) or MET (20 mg.kg-1 per day) for six weeks. Blood pressure was measured using carotid artery intubation with a MP150 polygraph. The media thickness, wall cross-sectional area and thickness / internal diameter ratio were calculated by morphometry. Vascular fibrosis was evaluated by Masson′s trichrome staining. The collagen and fibronectin expression in vascules was measured by Western blotting. RESULTS Compared with the sham group 〔(17.6±0.5)kPa〕, the mean arterial blood pressure in the model group〔(23.3±0.7)kPa〕 was significantly increased(P<0.05), but was lowered by DOX 〔(20.5±0.7)kPa〕 and MET 〔(19.0±0.4) kPa〕 (P<0.05). Moreover, HE staining showed that tunica media thickness, artery vessel area and thickness / inner diameter in the model group were increased by 39.5%, 46.4% and 27.0%(P<0.05), respectively. The tunica media thickness was decreased by 16.0% and 26.1%( P<0.05), respectively, the artery vessel area by 22.8% and 26.1%(P<0.05), respectively, and the thick-ness / inner diameter by 17.0% and 26.0%( P<0.05) when the rats were treated with DOX and MET. Masson staining showed that the collagen accumulation in vascules increased, suggesting that AAC induced fibrosis. Meanwhile, vascular fibrosis induced by AAC was also reduced by MET or DOX. Western blotting also proved that the increase of collagen and fibronectin induced by AAC could be attenuated by DOX and MET(P<0.05). CONCLUSION DOX and MET are effective in suppressing the role of norepi-nephrine in vassels, which can attenuate AAC-induced vassels remodeling by preventing the binding between norepinephrine and adrenoceptors.

MR fingerprinting (MRF) is a technology which can be used to quickly get quantitative images and post processing.The program of signal acquisitions,post-processing and virtualization of MRF are innovatively designed,being able to obtain a highly repeatable and multi-parametric quantification map in one scanning.MRF has shown various advantages,such as short scanning time,high fault tolerance and easy recognition of image data with computer,therefore having broad prospect in clinical application.The research progresses of MRF were reviewed in this article.