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Objective @#To explore and optimize the in vitro primary culture method of astrocytes in neonatal mouse cerebral cortex , which provides a better solution for the in vitro culture of astrocytes.@*Methods @#In order to optimize the in vitro culture method of mouse cerebral cortex astrocytes , 3 ⁃day⁃old C57BL/6J mouse cerebral cortex tissues were taken , meninges and blood vessels were removed , digested by pancreatic enzymes and centrifuged , andhigh⁃glucose dulbecco ′s modified eagle medium (DMEM) was added to form cell suspension , which was purified by differential adhesion method , cross hand method and constant temperature shaking method.The cells were inoculated in poly⁃D ⁃lysine⁃coated culture bottles with different culture densities , and the purity of astrocytes was determined by morphological ob servation and immunofluorescence staining.@*Results @#The cells were inoculated at a density of 5 × 106 cells per bottle with good effect and high activity. The purity of astrocytes reached 99% by using high sugar DMEM medium combined with differential adhesion method , cross hand method and constant temperature shaking method.@*Conclusion @#The primary culture method of astrocytes in mouse cerebral cortex is successfully established and optimized.
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【Objective】 Annexin A2 (annexin A2, Anxa2) has been reported to regulate bioactivity in various tumors cells. The purpose of this study was to investigate the correlation between the expression of Anxa2 protein and the proliferation and migration abilities of bladder cancer pumc-91 cells. 【Methods】 The ANXA2 sequence was amplified and inserted into the pcDNA3.1(+) vector in order to prepare the pcDNA3.1(+)-ANXA2 plasmid. PcDNA3.1 (+)-ANXA2 was transiently transfected into pumc-91 bladder cancer cells by lipofectamine 2000. Western blotting assay was performed to detect the expression of Anxa2 protein in the blank group, the control group transfected with pcDNA3.1(+), and the experimental group transfected with pcDNA3.1(+)-ANXA2 plasmid. The proliferation ability of pumc-91 cells was detected using Cell Counting Kit-8(CCK8), and the migration level of pumc-91 cells was detected by transwell assay. Differences in detection data among the groups were compared using one-way ANOVA or repeated measures ANOVA. 【Results】 The plasmid construction was successful and the sequencing was absolutely correct. Western blotting assay showed elevated Anxa2 protein expression level in the experimental group compared to the blank and control groups. CCK8 assay suggested that the number of proliferating pumc-91 cells was significantly higher in the experimental group than in the blank group (P<0.001) and the control group (P=0.001). Transwell assay also showed that the number of pumc-91 cells crossing the membrane was significantly higher in the experimental group than in the blank group (P=0.011) and the control group (P=0.027). 【Conclusion】 Our findings suggested that up-expression of Anxa2 may play a critical role in regulating proliferation and migration of bladder cancer pumc-91 cells.
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Objective:To investigate acute adverse events and pregnancy outcome after vaccination of inactivated COVID-19 vaccine in the first trimester of pregnancy.Methods:The retrospective-prospective cohort study was conducted among pregnant women of 11-13 +6 weeks of gestation who visited the obstetric clinics for prenatal check in Lianyungang Maternal and Child Health Hospital from May to November in 2021, after registration for perinatal health cards in the community. Those who met the inclusion criteria were recruited and were divided into vaccination group and non-vaccination group according to whether they received inactivated COVID-19 vaccine in the first trimester. Women in the vaccination group were further divided into 1-dose group and 2-dose group. Information, including pregnancy-related screening, pregnancy complications, pregnancy outcome and acute adverse events, were collected and compared with independent samples t-test or ANOVA, Kruskal- Wallis H test or Mann-Whitney U test, χ2 test or Fisher's exact probability method. Results:Totally, 105 pregnant women were analyzed in 1-dose group, 90 in 2-dose group, and 194 in non-vaccination group. (1) There were no statistically significant differences in the occurrence of acute adverse events [1-dose group: 2.86% (3/105); 2-dose group: 6.67% (6/90); non-vaccination group: 4.63% (9/194); χ2=1.59; vaccination group was 4.61% (9/195), when compared with non-vaccination group, χ2=0.00], abnormal pregnancy-related screening indicators and abnormal pregnancy outcome among the three groups (all P>0.05), neither between the vaccination and non-vaccination group (all P>0.05). The acute adverse events in these women included fever, pain at the inoculation site, fatigue, local induration and rash.(2) The differences in hypertensive disorders in pregnancy among the three groups were statistically significant [1-dose group: 10.5%(11/105); 2-dose group: 17.8%(16/90); non-vaccination group: 7.7%(15/194); χ2=6.46, P=0.040], and the incidence was higher in the 2-dose group than that in the non-vaccination group (adjusted by Bonferroni, P<0.017). (3) Regarding other pregnancy complications, no difference was found among the three groups (all P>0.05), neither between the vaccination and non-vaccination group (all P>0.05). Conclusion:The risk of acute adverse events and adverse pregnancy outcome is similar in pregnant women who received inactivated COVID-19 vaccine versus those who did not in the first trimester, and regular blood pressure monitoring is recommended for those who received two doses of inactivated COVID-19 vaccine.
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Objective@#To explore and optimize the primary culture method of neonatal mouse hippocampal neurons in vitro．To construct a G-protein-coupled receptor kinase 2 ( GRK2) knockout HT22 cell line．@*Methods @#Hippocampal tissue of C57BL6 /J mice on day 1－2 was taken，digested with trypsin and pipetted to form a cell suspension，and supplement was added to Neurobasal-A medium to maintain cell growth． CRSIPR / Cas9 gene editing technique was used to construct HT22-GRK2 －/ － cell line，and the knockout efficiency of GRK2 was detected by immunofluorescence staining and Western blot. @*Results @#Primary hippocampal neurons of newborn mice were put into six-well plates with 3 × 107 /well using a serum-free culture method，which could get a high purity and good activity ; HT22-GRK2 －/ － cell line was constructed successfully．@*Conclusion@#The primary culture method of mouse hippocampal neurons was successfully established and optimized，and HT22-GRK2 －/ － cell line was successfully constructed by CRSIPR / Cas9 gene editing technique.
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Background/Aims@#Single-balloon enteroscopy (SBE) has been widely used in diagnosing small bowel disease. We conducted this study to systematically appraise its technical and clinical performance. @*Methods@#Studies on SBE published by September 2018 were systematically searched. Technical and clinical performance data were collected and analyzed with descriptive or meta-analysis methods. @*Results@#In total, 54 articles incorporating 4,592 patients (6,036 procedures) were included. Regarding technical parameters, the pooled insertion depths (IDs) for anterograde and retrograde SBE were 209.2 cm and 98.1 cm, respectively. The pooled retrograde ID in Asian countries was significantly greater than that in Western countries (129.0 cm vs 81.1 cm, p<0.001). The pooled anterograde and retrograde procedure times were 57.6 minutes and 65.1 minutes, respectively.The total enteroscopy rate was 21.9%, with no significant difference between Asian and Western countries. Clinically, the pooled diagnostic yield of SBE was 62.3%. Obscure gastrointestinal bleeding (OGIB) was the most common indication (50.0%), with a diagnostic yield of 59.5%. Vascular lesions were the most common findings in Western OGIB patients (76.9%) but not in Asian ones (31.0%). The rates of severe and mild adverse events were 0.5% and 2.5%, respectively. @*Conclusions@#SBE is technically efficient and is clinically effective and safe, but total enteroscopy is relatively difficult to achieve with this technique. Etiologies of OGIB in Asian countries differ from those in Western countries.
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Objective:To reduce the occurrence of false alarm of vital signs during the use of ECG monitor by using stress ring to fix the lead wire of ECG monitor. Methods 200 patients who used ECG monitor in our department from March 2018 to February 2019 were intervened by two methods. The control group ( n=100) was connected and observed by routine method. The experimental group ( n=100) was added with lead wire to form "stress loop" on the basis of this method. Methods:The connection and fixation of ECG monitors were carried out. The differences of vital signs, heart rate/rhythm and respiratory false alarm between the two groups during ECG monitoring were compared, and the changes of nurses' workload, Hamilton anxiety score and satisfaction were compared.Results:There were significant differences in heart rate/rhythm and respiratory false alarm between the two groups ( P < 0.05). The scores of anxiety, nurses' workload and satisfaction in the experimental group were significantly improved. Conclusion:During the application of ECG monitor, the lead wire coiled into "stress ring" can effectively reduce the ECG monitor vital signs monitoring center rate/rhythm, respiratory false alarm phenomenon, reduce the workload of nurses, improve the satisfaction of nurses, and reduce anxiety of patients.
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In the Qing dynasty Cheng Zhongling based on the Cheng's Bixie-Fenqing formula of Yang Tan, a doctor in the Southern Song dynasty, retained the original Sichuan Bixie and Acorus tatarinowii, and add Phellodendron chinense, and Semen plantaginis to form Cheng's Bixie-Fenqing formula. It had the function of get rid of dampness and heat, and is a classic prescription for stranguria, like the treatment of white turbidity. The author retrieved the related literature of Cheng's Bixie-Fenqing formula in recent years, analyzed the composition and clinical application of Cheng's Bixie-Fenqing formula, and summarized the advantages and disadvantages of the existing researches, providing reference for the further development of Cheng's Bixie-Fenqing formula.
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In the Qing dynasty Cheng Zhongling based on the Cheng's Bixie-Fenqing formula of Yang Tan, a doctor in the Southern Song dynasty, retained the original Sichuan Bixie and Acorus tatarinowii, and add Phellodendron chinense, and Semen plantaginis to form Cheng's Bixie-Fenqing formula. It had the function of get rid of dampness and heat, and is a classic prescription for stranguria, like the treatment of white turbidity. The author retrieved the related literature of Cheng's Bixie-Fenqing formula in recent years, analyzed the composition and clinical application of Cheng's Bixie-Fenqing formula, and summarized the advantages and disadvantages of the existing researches, providing reference for the further development of Cheng's Bixie-Fenqing formula.
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OBJECTIVE: To investigate the prognostic factors of esophageal cancer with multiple organ metastases and establish a prognostic prediction model. METHODS: Patients data were extracted from the SEER database. The clinical data of 388 patients with esophageal cancer with multiple organ metastases were retrospectively analyzed. Risk factors were analyzed by log-rank method and survival curves were drawn by K-M method. Multivariate analysis was performed by Cox proportional hazard model to obtain independent prognostic factors for multi-organ metastasis of esophageal cancer. A prediction nomogram was further established.RESULTS: The mean survival time of patients in this study was 7.3 months, and the survival rates for 1-, 3-, and 5-year were 15.5%,1.2%, and 0, respectively. Age was an independent prognostic factor. The value of C-index was 0.618. CONCLUSION: The prognosis of esophageal cancer with multiple organ metastases is poor. Age at the diagnosis and patterns of multiple organ metastases are related to the survival time of patients. The prediction nomogram provided a good prognosis prediction.
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OBJECTIVE: To establish a prediction model for the prognosis of patients with esophageal cancer lung metastasis.METHODS: Data from 194 patients with esophageal cancer lung metastasis from 2010 to 2015 was collected from the National Cancer Institute Surveillance, Epidemiology and End RESULTS:(SEER) database. The best cutoff value for age was determined by X-tile software.Prognostic factors were analyzed by SPSS(v25.0) with the log-rank method and the Cox proportional hazard model. Risk factors from univariate analysis were used to construct prediction nomogram with R studio software(version 3.5.1). RESULTS: The median survival time of 194 patients with esophageal cancer lung metastasis was 7.0 months, the 3-month survival rate was 69.9%, and the 1-year survival rate was 27.7%. Age(HR=1.51, 95% CI: 1.066-2.140) and pathological type(HR=0.736, 95% CI: 0.543-0.998) were independent prognostic factors for patients with esophageal cancer lung metastasis. The value of C-index was 0.634(95% CI=0.585-0.683). CONCLUSION: For patients with esophageal cancer lung metastasis, being young and adenocarcinoma are associated with a better prognosis. The prediction of the nomogram is good.
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OBJECTIVE: To analyze the prognostic factors related to liver metastasis of esophageal cancer and establish an effective prediction model. METHODS: The data of 464 cases of esophageal cancer with liver metastasis from 2010 to 2015 was collected from the National Cancer Institute SEER database by SEER stat 8.3.5 software. SPSS(v25.0) was used to analyze the prognostic factors of esophageal cancer liver metastasis and Kaplan-Meier curve was used for survival analysis. We introduced the meaningful variables of single factor analysis in Cox proportional hazard model and multivariate analysis and obtained the independent influencing factors of prognosis.Independent factors were then included in the accelerated failure time model to construct the nomogram. RESULTS: The mean survival time of patients in this study was 11.6 months(95%CI: 10.075-13.209), and their 1-, 3-, and 5-year survival rates were 29.4%, 5.5%, and 0,respectively. Age(HR=1.452, 95% CI: 1.175-1.795), marriage(HR=0.753, 95%CI: 0.611-0.927) and surgery(HR=0.428, 95% CI: 0.227-0.807) were independent prognostic factors for patients. We constructed the nomogram with risk factors of prognosis, and the C-index value was 0.614. CONCLUSION: The prognosis of esophageal cancer liver metastasis is poor. being young, Being married, and surgery are associated with better survival, and the nomogram we have constructed is proved to have good predictive ability.
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OBJECTIVE: To construct a model for predicting the prognosis of esophageal cancer bone metastasis. METHODS: The clinical data of 183 patients with esophageal cancer bone metastasis were analyzed retrospectively, and the prognostic factors of patients were analyzed by log-rank method and Cox proportional hazard model. Nomogram was constructed with the accelerated failure-time model.RESULTS: The average survival time(10.0 months, 95% CI:7.758-12.338) of patients aged 28-70 years was longer than that of patients aged 71-91(6.4 months, 95% CI:4.365-8.428)(χ~2=4.077, P=0.043). The prognosis of unmarried patients(average 7.0 months) was worse than that of the married(10.5 months on average)(χ~2=12.841, P<0.001). As for prognoses of different pathological types, the differences between adenocarcinoma(average 10.2 months, 95% CI:7.797-12.548), squamous cell carcinoma(average 6.4 months,95%CI:3.895-8.899) and other types(average 4.0 months, 95% CI:4.000-4.000) were statistically significant(χ~2=7.171, P=0.028).There were also significant differences between the prognoses of patients with different T stage(χ~2=8.833, P=0.032). Nomogram was constructed with the risk factors above and the C-index reached 0.675(95%CI: 0.626-0.725). CONCLUSION: The prognosis of esophageal cancer bone metastasis was poor. Marriage, T stage, histological grade and pathological types were risk factors affecting prognosis, while N stage didn't appear to show obvious effect on prognosis. The nomogram was tested to have a good predictive capacity.
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OBJECTIVE: To investigate the prognostic factors of esophageal cancer with brain metastasis. METHODS: SEER Stat 8.3.5 was used to collect 39 cases of esophageal cancer with brain metastasis from 2010 to 2015 in the Surveillance, Epidemiology and End RESULTS:(SEER) database. X-tile software was used to determine the best cut-off value of the age. Prognostic factors were analyzed with log-rank and Cox proportional hazard model by SPSS(v25.0). RESULTS: The median survival time of patients with esophageal cancer with brain metastasis was 7.0 months, the 6-month survival rate was 53.3%, and the 1-year survival rate was 16.3%. Only age(χ~2=4.045, P=0.044)was the prognostic factor, while there was insufficient evidence to show whether gender, marriage, race, primary site, histological grade,surgery, pathological type, T stage or N stage was associated with the prognosis of the patients. CONCLUSION: Brain metastasis is a rare metastatic type of esophageal cancer. Age is associated with worse prognosis, while the influences of other risk factors are not clear.Active treatment can lead to better prognosis.
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Esophageal cancer is a common malignant tumor of the upper gastrointestinal tract. Early symptoms of the disease are inconspicuous and the disease is often diagnosed at a later stage, leading to higher morbidity and mortality. Esophageal cancer morbidity and mortality in both genders ranks among the top 10 most common cancers. Early detection and early treatment are effective means to reduce the incidence and mortality of esophageal cancer. Tumor markers play an important role in early diagnosis, treatment monitoring and prognosis evaluation of esophageal cancer. This paper reviews the clinical application of tumor markers related to esophageal cancer and the exploration and application progress of new tumor markers for esophageal cancer.
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Objective To investigate the value of serum uric acid and brain natriuretic peptide in the evaluation of progressive ischemic stroke.Methods Totally 63cases of progressive ischemic stroke patients in our hospital from October 2016to October 2017were selected as the observation group.According to the severity of the disease, the patients were divided into mild group[National Institute of Health Stroke Scale (NIHSS) score＜7], medium group (NIHSS score 7-15) and severe (NIHSS score＞15).Meanwhile, 60cases of nonprogressive ischemic stroke patients in the hospital from January 2014and December 2016were selected as the control group.After admission, 3 mL fasting peripheral venous blood was collected in each subject in both groups, and the serum was isolated.Enzyme-linked immunosorbent assay (ELISA) was used to determine the level of brain natriuretic peptide and uric acid/eroxidase coupling method was used to determine the level of serum uric acid.Results The levels of serum uric acid and brain natriuretic peptide were significantly higher than those of the control group (P＜0.05).The levels of natriuretic peptide and serum uric acid in severe group were significantly higher than those of mild group and medium group, and the levels of two indicators in medium group were significantly higher than those of mild group (P＜0.05).In addition, the sensitivity and specificity of combined detection were significantly higher than the single index detection of serum uric acid and brain natriuretic peptid (P＜0.05).Conclusion The combined detection of serum uric acid and brain natriuretic peptide have higher sensitivity and specificity.
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Sperm cryopreservation has been widely used in assisted reproduction, but conventional techniques are not suitable for the cryopreservation of small numbers of sperm. The application of the single sperm cryopreservation technique has significantly improved the clinical treatment of cryptozoospermia and non-obstructive azoospermia. Ever since Cohen et al first developed the method of single sperm cryopreservation in 1997, constant efforts have been made to develop the carriers for this technique. In this review, we mainly discuss the existing methods and clinical outcomes of single sperm cryopreservation.
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Humanos , Masculino , Azoospermia , Terapéutica , Criopreservación , Métodos , Heterocigoto , Oligospermia , Terapéutica , Reproducción , Preservación de Semen , Métodos , EspermatozoidesRESUMEN
Objective To investigate the inhibitory effects of radioactive 125I seeds on the growth and apoptosis of human lung adenocarcinoma cells A 549 in nude mice.Methods Human lung adenocarcinoma A549 cells were cultured in vitro and subcutaneously transplanted in BALA/c nude mice.When the tumor size reached(300 ±50)mm3,40 tumor-bearing mice were divided into 4 groups by the random number table method as 0,0.6,0.8 mCi(1 Ci＝3.7×1010Bq)groups and blank control group,with 10 in each group.The 125I seeds of 0,0.6,and 0.8 mCi were implanted into the transplanted tumors in nude mouse,respectively.The blank control group received no treatment.The weight of nude mice was measured regularly every 4 days.The mice were sacrificed on the 32 days after 125I seeds implication.The transplanted tumors were weighed and the weight gain curve for nude mice was plotted.Hematoxylin-eosin(HE)staining was used to observe the histopathological changes of the tumor tissue.Cell apoptosis was detected by TUNEL assay,and the expressions of the P21,Caspase-9,Survivin and Livin proteins were detected by immunohistochemical assay.Results There was no nude mice dead in each group.On the day 28 and 32 after 125I seeds treatment,the body weights of nude mice of 0.6 and 0.8 mCi groups became lighter than those of the blank control group(q＝4.26,9.19,4.11,11.59,P＜0.05),the tumor weights of the 0.6 and 0.8 mCi groups were significantly decreased(q＝5.021,5.692,P＜0.05)with tumor inhibition rates of about 49％and 62％.In the 0.6 and 0.8 mCi groups,a large number of tumor cells degenerated to be necrotic cells.In addition,the apoptotic indexes were(50.00 ±2.58)％and(62.33 ± 4.51)％in the 0.6 and 0.8 mCi groups,respectively,and higher than that of blank control group(27.00 ±4.69)％.The expressions of P21 and Caspase-9 proteins in the 0.6 and 0.8 mCi groups were significantly higher than that in the blank control group(χ2＝11.380,24.310,11.380,20.376,P＜0.05).The expressions of Survivin and Livin proteins in the 0.6 and 0.8 mCi groups was significantly lower than that in the blank control group(χ2＝9.643,23.254,15.429,26.667,P＜0.05).Conclusions Radioactive 125I seeds can inhibit the proliferation of tumor cells and promote the apoptosis of A 549 cells probably by up-regulating the expressions of P21 and Caspase-9 but down-regulating the expressions of Survivin and Livin.
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Objective To investigate the changes of hemorheological indexes in patients with intracranial infection after traumatic brain injury.Methods 87 cases with intracranial infection after brain trauma ,admit-ted in South Hospital of the Sixth People's Hospital Affiliated to Shanghai JiaoTong University ,from October 2016 to October 2017 ,were selected as the intracranial infection group ,and 80 cases without intracranial infec-tion after brain trauma ,admitted in the hospital from June 2015 to December 2016 ,were selected as the unin-fected group.hemorheological indexes and Glasgon Coma Scale (GCS) score were compared between the two groups.Results The high shear viscosity ,low shear viscosity ,plasma viscosity and fibrinogen(Fib) content in the intracranial infection group were higher than those in the uninfected group ,and the differences were statis-tically significant (P＜0.05) ;the GCS score of the intracranial infection group was lower than that of the un-infected group ,and the differences were statistically significant (P＜0.05) ;the high shear viscosity ,low shear viscosity ,plasma viscosity and Fib content in severe infection group were higher than those in moderate infec-tion group and mild infection group ,and the high shear viscosity ,low shear viscosity ,plasma viscosity and Fib content in moderate infection group were higher than those in mild infection group ,and the differences were statistically significant (P＜0.05) ;the GCS score of severe infection group was lower than those of moderate infection group and mild infection group ,the GCS score of moderate infection group was lower than that of mild infection group ,and the differences were statistically significant (P＜0.05).Conclusion Abnormal hem- orrheology is present in patients with intracranial infection after traumatic brain injury ,and its change is close-ly related to the severity of the disease.
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Objective To analyze the characteristics of dynamic contrast enhanced MR imaging (DCE-MRI)in prostate cancer (PCa)at 3.0T,and to evaluate the diagnostic value of DCE-MRI.Methods 85 patients with suepected PCa received conventional MRI and DCE-MRI.The signal intense-time (SI-T)curve was analyzed.Then the time to maximum (Tmax),the maximum degree of enhancement (STmax%),and the rate of enhancement (Rmax)were calculated.The differences of styles of SI-T curve and the parameters between the positive and negative group were compared respectively.Results 59 cases of PCa were proved by biopsy,and there was no evidence of tumor in 26 cases.507 zones had histopathological results with 250 zones in positive group and 257 zones in negative group .The most common style of SI-T curve in positive group was rapidly ascending followed with descending curve,the most common style of SI-T curve in negative group was persistent ascending curve and plateau curve.The mean values of Tmax,SImax%,Rmax were (69.49±22.53)s,1.74±0.43,7.83±3.80 in positive group respectively,while (175.61±52.64)s,1.05±0.35,1.86±1.10 in negative group respectively,there were statistically significant differences between the two groups(t =-24.24,1 6.34,1 7.75,P <0.01)respectively. The mean values of Tmax,SImax% ,Rmax were (8 9 .1 9 ± 3 1 .7 2 )s,1 .5 8 ± 0 .4 6 ,5 .2 1 ± 3 .3 4 in the low-risk group (Gleason score 2 - 6 )respectively,while (64.25±14.68)s,1.76±0.43,8.25±3.70 in the high-risk group (Gleason score 7-10)respectively, there were statistically significant differences between them(t = 7.09,-8.74,- 7.83,P <0.01).Conclusion 3.0T DCE-MRI has great value in the diagnosis of PCa.
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Acute myocardial infarction is one of the major causes of death in the world. Research on animal models of myocardial infarction has a great significance in improving the pathophysiology, diagnosis and treatment of human myocardial infarction. At present, problems such as low survival rate and complicated preparation still exist. In recent years, some progress has been made in the preparation of animal myocardial infarction models, which can improve the survival rate, qualified rate and speed. This paper analyzes the preparation of rat models of acute myocardial infarction and related research, combined with actual factors to simplify the process of preparation and improve the skills of scientific researchers, so as to provide a source of stable and reliable acute myocardial infarction models for medical research.