BACKGROUND: The observation of vascular endothelial growth factor gene expression of cerebrovascular diseases and ultrastructure of cells may be helpful to understand angiogenesis and its relative cellular factors involved in the pathogenesis at cellular and molecular levels. OBJECTIVE: To investigate the method of culture of human cerebral cap illary endothelial cell by separation of capillary fragment in vitro, and to ob serve vascular endothelial growth factor gene expression and ultrastructure of cells. DESIGN: A randomized controlled research on technique and method. SETTING: The neurosurgery department of a general hospital of a military area command of Chinese PLA and the neurosurgery department of a college hospital. PARTICIPANTS: Eighteen patients with arteriovenous malformation of brain(Spetzler Ⅱ-Ⅲ grade), as confirmed by aortocranial angiography before operation, in the Department of Neurosurgery, General Hospital of Shenyang Military Area Command of Chinese PLA were included. The material was obtained from fresh integrated specimen of arteriovenous malfor mation of brain with surrounding fresh brain tissues during the opera tion. Capillary endothelial cell was separated by homogenate, filtration and enzymatic digestion techniques. Cells grew well in culture flask and were divided into 4 groups(hypoxia state for 2, 4, 8 hours groups and control group). Each group contained four flasks.METHODS: Simulation of anoxia condition: volume faction 0.95 N2 and volume fraction 0.05 CO2. Expression of factor Ⅷ related antigen in cells was detected by immunohistochemistry. mRNA expression of vascular endothelial growth factor on endothelial in every group was observed by RT-PCR, protein content of vascular endothelial growth factor in supernatant detected by enzyme-linked immunoadsordent assay, and cellular ultrastructural change observed by transmission electron microscopy.MAIN OUTCOME MEASURES: mRNA expression of vascular endothelial growth factor on endothelial cell and protein content of vascular endothelial growth factor in supernatant in control group and every hypoxia groups; cellular ultrastructural changes.RESULTS: Under phase contrast microscope, cultured living cells had mono-layer pebble-like typical character. More than 90% of were factor Ⅷrelated antigen(FⅧ-RA) staining positive. mRNA and protein expression of vascular endothelial growth factor in hypoxia 4 hours group was 0.98 ±0. 19,( 180. 77 ± 20. 15) ng/L, which was significantly higher than in control group [0, (26. 20 ± 6.33) ng/L, P ＜ 0.01 ]. Eight hours later, expression decreased [(0. 35 ±0.07), (31.68 ±8.34) ng/L]; swollen mitochondrion, dilated endoplasmic reticulum, and lysosome vesiculation were found.CONCLUSION: Humane cerebral capillary endothelial cell can be cultured by separation of capillary fragment, which is easy to operate and the cellular purity is reliable. In the early stage of ischemia and hypoxia, expression of vascular endothelial growth factor is not enough to maintain cellular ultrastructure integrity. Cells may be injured along with the prolong of hypoxia.Zhao MG, Tang T, Gao YZ, Pu PY, Wei XZ. Culture of human cerebral capillary endothelial by separation of capillary fragment and the observation of vascular endothelial growth factor gene expression and cell ultrastructure. Zhongguo Linchuang Kangfu 2005; 9(21):211-3 (China) [www. zglckf. com]

Objective To investigate the gene expression of vascular endothelial growth factor(VEGF) and ultrastructural changes in cultured endothelial cells from human cerebral microvessels under hypoxic conditions.Methods Human cerebral microvessels were isolated from freshly obtained specimens of normal brain adherent to resected cerebral arteriovenous malformations.The expression of factor Ⅷ-relative antigen(FⅧ-RA) in cultured cells was observed with immunocytochemistry.The level of VEGFmRNA in cells and released VEGF protein in cell supernatant were determined by RT-PCR analysis and ELISA respectively when they were exposed to hypoxic conditions(95% N_2,5% CO_2;two hours,four hours,eight hours) or maintained in basal condition.Ultrastructural changes in cells were also observed by electron microscopy.Results In inverted microscope the cultured cells showed contact inhibition and a rounded cobblestone appearance.More than 90% of them were stained strongly with antibodies against FⅧ-RA.Significant VEGF mRNA and protein accumulated when these cells were exposed to hypoxia for 4 hours.However,their VEGF expression was down-regulated after hypoxia for 8 hours and a number of vesicles and swollen mitochondria were present in the cytoplasm.Conclusion The level of VEGF expression may havesignificant relationship with ultrastructural changes in human cerebral endothelial cells under hypoxic conditions.

adder irritation after intra-vesical sodium hyaluronate therapy. Conclusion Intravesical sodium hyaluronate therapy can effec-tively relieve clinical symptom of IC and improve IC patients' quality of life and is well tolerated.

Objective To explore the signaling pathway of apoptosis induced by Paclitaxel (PTX) in leukemia cells and the chemosensitizing effect of adding short hairpin RNA(shRNA) on PTX,which targets the silencer of death domains(SODD).Methods After being treated with PTX,the expressions of SODD,B-cell lymphoma/leukemia-2 (Bcl-2),nuclear factor kappa B (NF-κB) and Caspase-3 proteins in Jurkat cells were determined by Western blot ;the shRNA-SODD vectors were constructed and transfected into Jurkat cells by electroporation,and then G418 was used to select the stable tranfected cell line expressing the shRNA-SODD recombinant plasmids.The incidence of cell apoptosis induced by PTX was determined by flow cytometry labeled with propidium iodide.Results During the process of inducing apoptosis of Jurkat cells,PTX could significantly down-regulate the expressions of SODD and Bcl-2 proteins,degrade Caspase-3 and activate NF-κB.The apoptotic sensibility of Jurkat cells transfected with shRNA-SODD to PTX was significantly increased compared with the control group,and the difference was statistically significant (F =10.35,P ＜ 0.05).Conclusions PTX can effectively induce apoptosis of Jurkat cells.Perhaps,SODD/Bcl-2 represents a specific apoptotic signaling pathway of PTX in leukemia cells and this apoptotic signaling pathway is Caspase-3-dependent,in which the function of NF-κB is to modulate the correlative apoptotic factors.Inhibiting the expression of SODD through transfecting shRNA-SODD vectors can significantly increase the apoptotic sensibility of leukemia cells to PTX.

OBJECTIVE: Analysis of the correlation between telomerase and expression of its related proteins may provide insight into the molecular mechanism of nasopharyngeal carcinogenesis. Here, we investigate the effect of short hairpin RNA (shRNA) specific for human telomerase reverse transcriptase (hTERT) mRNA on the expression of proliferating cell nuclear antigen (PCNA) and Caspase-3 in nasopharyngeal carcinoma (CNE) cells. METHOD: shRNA expression vectors targeting the mRNA of hTERT were constructed. Cells were treated with the constructed expression vectors and telomerase activity was measured by telomeric repeat amplification ELISA (TRAP-ELISA). Cell viability was examined using the MTT assay. Cell apoptosis was detected by TUNEL method. The expression of PCNA and Caspase-3 proteins, was determined by Western blotting. RESULT: shRNA specific for hTERT mRNA significantly inhibited telomerase activity, suppressed cell viability and induced apoptosis in CNE cells. In addition, the expression of PCNA was inhibited, while the expression of Caspase-3 was up-regulated. CONCLUSION Our results suggest that shRNA directed against hTERT inhibits cell viability by regulating telomerase activity and its related protein expression in NPC cells. Therefore, RNA-interfering technology may be a promising strategy for the treatment of nasopharyngeal cancer.
Caspase 3 ; metabolism ; Cell Line, Tumor ; Cell Survival ; Humans ; Nasopharyngeal Neoplasms ; genetics ; metabolism ; Proliferating Cell Nuclear Antigen ; metabolism ; RNA, Small Interfering ; Telomerase ; genetics ; Transfection

Objective:To evaluate the effect of transnasal humidified rapid insufflation ventilatory exchange (THRIVE) on regional cerebral oxygen saturation (rScO 2) during induction of general anesthesia in patients undergoing traumatic brain injury (TBI) emergency surgery. Methods:A prospective randomized controlled trial was conducted. The TBI emergency general anesthesia patients who underwent intracranial hematoma removal surgery at the Northern Jiangsu People's Hospital from January to July in 2023 were enrolled. The patients were divided into a conventional mask ventilation group and a THRIVE group using a random number table method. The patients in the conventional mask ventilation group were anesthetized and induced to pre oxygenate without positive pressure ventilation in the front mask for 10 minutes, with an oxygen flow rate of 8 L/min and an fraction of inspired oxygen (FiO 2) of 1.00. After anesthesia induction for about 90 s, tracheal intubation was performed after the muscle relaxant took effect (patient's jaw muscle was relaxed). The patients in the THRIVE group were pre oxygenated with THRIVE for 10 minutes, with an oxygen flow rate of 30 L/min and a FiO 2 of 1.00. During anesthesia induction, the oxygen flow rate was increased to 50 L/min, and anesthesia induction medication was used. The lower jaw of patient was supported with both hands to maintain airway patency, and the patient's mouth was kept closed throughout the process. After the muscle relaxant took effect (the patient's jaw muscle was relaxed), tracheal intubation was performed. At the time of patient entering the operating room, 10 minutes of pre oxygenation, and immediately after successful intubation, rScO 2 was measured on the surgical and non-surgical sides. At the same time, ultrasound was used to measure the cross-sectional area (CSA) of the gastric antrum and arterial blood gas analysis was performed. The partial pressure of end-tidal carbon dioxide (P ETCO 2) during the first mechanical ventilation after successful tracheal intubation, the incidence of hypoxemia [pulse oxygen saturation (SpO 2) < 0.95] during tracheal intubation, as well as prognostic indicators such as the length of intensive care unit (ICU) stay, total length of hospital stay, and Glasgow outcome scale (GOS) score at discharge were recorded. Results:During the study period, a total of 70 TBI patients underwent emergency general anesthesia surgery, of which 2 patients died postoperatively, 2 patients were unable to cooperate with closed mouth breathing, and 3 patients had poor ultrasound image acquisition in the gastric antrum, all of whom were excluded. A total of 63 patients were ultimately enrolled, including 32 in the conventional mask ventilation group and 31 in the THRIVE group. There were no statistically significant differences in gender, age, body mass index (BMI), American Society of Anesthesiologists (ASA) classification, Glasgow coma scale (GCS) score, optic nerve sheath diameter (ONSD), baseline vital signs, fasting situation, anesthesia time, surgical time, and intraoperative blood loss between the patients in the two groups, indicating comparability. When entering the operating room, there was no statistically significant difference in rScO 2 on the surgical and non-surgical sides, and blood gas analysis indexes arterial partial pressure of oxygen (PaO 2) and arterial partial pressure of carbon dioxide (PaCO 2) between the patients in the two groups. When pre oxygenated for 10 minutes, both the surgical and non-surgical sides rScO 2 levels in the THRIVE group were significantly higher than those in the conventional mask ventilation group (surgical side: 0.709±0.036 vs. 0.636±0.028, non-surgical side: 0.791±0.016 vs. 0.712±0.027, both P < 0.01), and the PaO 2 was significantly increased [mmHg (1 mmHg≈0.133 kPa): 450.23±60.99 vs. 264.88±49.33, P < 0.01], PaCO 2 was significantly reduced (mmHg: 37.81±3.65 vs. 43.59±3.76, P < 0.01), and the advantage continues tilled immediately after successful intubation. There was no statistically significant difference in CSA at each time point of ultrasound examination between the two groups. Compared with the conventional mask ventilation group, the patients in the THRIVE group showed a significant decrease in P ETCO 2 during the first mechanical ventilation after successful tracheal intubation (mmHg: 43.10±2.66 vs. 49.22±3.31, P < 0.01), and the incidence of hypoxemia during tracheal intubation was also significantly reduced [0% (0/31) vs. 28.12% (9/32), P < 0.01]. In terms of prognostic indicators, there was no statistically significant difference in the length of ICU stay and total length of hospital stay between the patients in the conventional mask ventilation group and the THRIVE group [length of ICU stay (days): 10 (9, 10) vs. 10 (9, 11), total length of hospital stay (days): 28.00 (26.00, 28.75) vs. 28.00 (27.00, 29.00), both P > 0.05]. However, the proportion of patients in the THRIVE group with a good prognosis at discharge (GOS score > 3) was significantly higher than that in the conventional mask ventilation group [35.5% (11/31) vs. 12.5% (4/32), P < 0.05]. Conclusion:THRIVE can significantly increase rScO 2 during anesthesia induction in TBI emergency surgery patients and improve their neurological function prognosis.

Objective:To explore the clinical characteristics of patients with cholelithiasis in Jinshan district of Shanghai city. Methods:The clinical characteristics of 3455 patients with cholelithiasis treated in Jinshan Branch of Shanghai Sixth People′s Hospital from Jul 2010 to Dec 2012 were analyzed.Results:Among them,3127 patients (90.5%)had clinical symptoms,and 328 patients (9.5%)had no symptoms.The incidence rate of symptomatic cholelithiasis in male patients was lower than that in female patients,while the incidence rate of asymptomatic cholelithiasis in male patients was higher than that in female patients (P <0.05).The incidence rate of asymptomatic cholelithiasis in patients decreased with the increase of age,and the difference was statistically significant(P <0.05).The incidence of cholecystolithiasis was 54.5%.The appearance of symptoms in patients with cholelithiasis was not correlated with the body mass index(BMI)and the location of cholelithiasis(P >0.05).Conclusions:Jinshan district of Shanghai city has high incidence of cholelithiasis.The appearance of Symptoms in patients with cholelithiasis is correlated with gender and age,but not correlated with BMI and location of cholelithiasis.

Objective To provide more reference base for the prevention and treatment of acute cerebral infarction through the correlation study between the onset of acute cerebral infarction and meteorological factors during the same period in Dalian area.Methods From January 1,2015 to December 31,2015,the data of hospitalized medical records and meteorological data during the same period in 3 5 7 8 consecutive patients with acute cerebral infarction admitted to three tertiary hospitals in Dalian were analyzed retrospectively.The seasonal regularity of acute cerebral infarction in Dalian area was analyzed descriptively.A single retrospective 1 ∶ 1 matched case-crossover study design was used to analyze the effect of mean air temperature lag 0-3 days on the onset of acute cerebral infarction.Pearson correlation analysis was used to analyze the influence of meteorological factors of the day on the onset of acute cerebral infarction,at the same time,the relationship between stroke subtypes and meteorological factors was investigated.Results (1) The incidence of acute cerebral infarction was higher in winter (December,January,and February) and it was lower in summer (from June to August).(2) Hysteresis analysis:when the temperature was reduced by 1 ℃each time,the OR values of acute cerebral infarction lagging behind 0 and 1 d were 1.034 (95％ CI 1.012-1.056) and 1.025 (95％ CI 1.008-1.042) respectively.There was significant difference (all P ＜ 0.05).The correlation between the temperature of the day of onset and the onset of acute cerebral infarction was the highest.(3) The onset of cerebral infarction was negatively correlated with the average temperature of the day and hours of sunshine (r =-0.392,-0.260,all P ＜ 0.01),and it was positively correlated with the daily average air pressure (r =0.403,P ＜ 0.01).Among them,the correlation of the type of cardiogenic embolism and each meteorological factor was the highest.The correlation coefficients of daily average temperature,daily air pressure,and hours of sunshine were-0.836,0.733,and-0.629,respectively (all P ＜ 0.01).Conclusion A cold and high air pressure may trigger the onset of acute cerebral infarction,especially cardiogenic cerebral infarction.

Objective To analyze the layout and shielding effectiveness of medical accelerator vaults, and to provide a reference for the layout, shielding design, and optimization of protection of medical accelerator vaults. Methods Four medical accelerator radiotherapy vaults were selected. The layouts of these vaults were compared with the layout requirements in the radiation therapy protection standards. For each vault, the dose rates at four points of interest outside the shielding were calculated, including the primary shielding area, secondary shielding area, maze outer wall, and lateral shielding area. These values were then compared with the actual measurements obtained using a dose rate meter. Results All four vaults were located on the ground floor of the building and included a maze, with the auxiliary rooms all placed outside the treatment rooms. However, one vault was not located at one end of the building, and in another vault, the control room was exposed to direct irradiation of the useful beam. The calculated dose rates outside the primary shielding area ranged from 0.04 μSv/h to 0.62 μSv/h, while the measured values ranged from 0.10 μSv/h to 0.66 μSv/h, with the measured values being higher than the calculated ones. The calculated dose rates outside the secondary shielding area ranged from 0.0005 μSv/h to 0.12 μSv/h, those outside the maze outer wall ranged from 0.06 μSv/h to 0.18 μSv/h, and those outside the lateral shielding area ranged from 0.009 μSv/h to 0.15 μSv/h; the measured values were all below the detection limit. Conclusion Inadequate layouts were observed in some of the accelerator vaults. The calculated and measured dose rates at points of interest outside the vaults were both significantly lower than the shielding protection requirements in standards, indicating that the shielding designs were overly conservative. Construction units should follow the principle of optimization of radiation protection to design the layout and shielding reasonably and pay attention to the underestimation of dose rates outside the primary shielding caused by the tenth value layer.

Objective To explore the outcomes and complications of 111 patients with pituitary neoplasms after neuroendoscopic endonasal transsphenoidal surgeries.Methods A retrospective review of clinical and radiographic data of 111 patients (113 procedures),admitted to our hospital from June 2010 to July 2014,was performed.The resection rate,clinical cured rate and complications in pituitary neoplasms of different Knosp classifications were compared.Results Gross total resection (GTR) rate for pituitary neoplasms according to Knosp classification was 97.4％ in grade 0,85.7％ in grade 1,88.7％ in grade 2,51.9％ in grade 3 and 0％ in grade 4,with significant differences (P＜0.05);GTR rate in pituitary neoplasms of grade 3 and 4 was significantly lower than that in pituitary neoplasms of grade 1 and 2 (P＜0.05).Clinical cured rate of the prolactin adenomas of different Knosp grades was significantly different,and that of grade 0 was statistically higher than that of grade 1-4 (P＜0.05).Clinical cured rate of growth hormone adenoma,adeno-corticotrophic stimulating hormone adenoma and mixed-functional adenomas of different grades showed no significant differences (P＞0.05).Chief complaints were improved in 75.0％-91.7％ non-functioning adenomas.Postoperative severe complications included 3 patients (2.7％) with cerebrospinal fluid rhinorrhea,4 (3.5％) intracranial hematoma,1 (0.9％);postoperative mild complications included 3 (2.7％) epistaxis,14 (12.4％) transient diabete insipidus,1 (0.9％) permanent diabetes insipidus and 3 (2.7％) hyponatremia.Mortality was observed in one patient with intracranial hematoma.Conclusions Neurendoscope provides a superior view of the sellar region,and improves the extent of resection of pituitary adenoma especially for those with cavernous sinus invasion;however,blind pursuit of GTR for tumor with Knosp classification greater than grade 3 will increase the risk of severe complications.Safety should always be the priority for a procedure with a long learning curve like neuroendoscopic surgery.Remission of residue tumor can be addressed by subsequent radiotherapy/stereotactic surgery.