The muscarinic receptor family expressed in smooth muscle throughout the body is thought to be composed of five subtypes coupling to distinct signaling systems,respectively.The population in smooth muscle is composed of mainly M 2 and M 3 subtypes in a 80% to 20% mixture. The muscarinic receptor, mainly M 3 receptor, play an important role in regulating gastrointestinal smooth muscle contraction. Selective muscarinic M 3 antagonist should have therapeutic utility in the treatment of gastrointestinal disease.

Objective To investigate effect of dicoumarol on the expression of serum tumor necrosis factor-α( TNF-α) in patients with acute cerebral infarction.Methods Eighty cases patients with acute cerebral infarction were admitted and randomly divided into two groups by digital draw:control group and observation group, 40 cases in each group.The control group were given conventional therapy for treatment of acute cerebral infarction, and observation group were given dicoumarol thrombolysis on the basis of control group.The serum TNF-αlevel and neurologic impairment score were compared between two groups pre-and post-treatment.Results After treatment of 6 h, the serum TNF-αlevel in observation group was significantly lower than that in control group (P<0.05).After treatment of 24 h, 48 h and 30 d, there were no significant differences of serum TNF-αlevels between two groups.After treatment of 5 d, 14 d and 28 d, the neurological deficit scores in observation group were significantly higher than those in control group (P<0.05).Conclusion Dicoumarol could effectively reduce TNF-αlevels in serum and extent of cerebral cell damage in patients.

Objective To observe the effect of pretreatment of aprotinin on nitric oxide (NO) and nitric oxide synthase (NOS) contents after ischemia-reperfusion injury of spinal cord in rabbits.Methods 45 rabbits were randomly divided into aprotinin treatment group (group A), normal saline control group (group B) and pseudo-surgical operation group (group C) with 15 rabbits in each group. The infrarenal segment in abdominal aorta was clamped for 60 min to construct the model of lumbosacral spinal cord ischemia in rabbits. Reperfusion was followed and kept on for 24 h until the blood flow regained normal. Aprotinin was given 3×107 IU/kg as a short time intravenous injection for 10 min before ischemia, and then was drilled with micro pump by 1×107 IU/kg/h. Normal saline was used in group B, the ischemia-reperfusion duration between group A and group B remained same. The group C was only exposured abdominal aorta and not clamped. The rabbits were killed before ischemia and at 8 h, 24 h after ischemia-reperfusion, lumbar segment spinal cords were harvested to detect contents of NO and NOS of spinal cord.Results After 8 h of ischemia-reperfusion,the contents of NO, total NOS (TNOS), and induced NOS (iNOS) in group A and group B were more than that before ischemia (P＜0.05). After 8 h of ischemia-reperfusion, there was a significant difference in the contents of NO, TNOS, iNOS between group A and group B (P＜0.05～0.01). After 24 h of ischemia-reperfusion, there was a significant difference too between group A and group B (P＜0.01). After 8 h and 24 h ischemia-reperfusion, the contents of NO, TNOS, iNOS in group A and group B were more than that in group C (P＜0.01).Conclusion During the ischemia-reperfusion, more NO produced is an important factor of spinal cord injury. Aprotinin can decrease the contents of NO and ischemia-reperfusion injury to spinal cord of rabbits.

Objective To investigate the enteral nutrition(EN)complications in ICU critically ill patients and their risk factors.Methods The relevant information of 376 EN patients from three tertiary hospitals in Qingdao from September 2015 to April 2016 were retrospectively analyzed.The incidences of EN complications and their risk factors were analyzed.Results In these 376 patients,the complications included diarrhea(n=117,31.12％),constipation(n=89,23.67％),and reflux(n=102,27.13％).The incidence of diarrhea showed significant difference in terms of the application of antibiotics(OR=3.55),age(OR=1.03),fasting(OR=4.07),oral potassium preparations(OR=2.91),nasal feeding rate(OR=1.03),and nasal feeding volume(OR=1.002).The incidence of constipation showed significant difference in terms of nasal fluid type(OR=7.79),analgesics(OR=10.34),age(OR=1.06),nasal feeding rate(OR=0.95),and nasal feeding volume(OR=0.97).The incidence of reflux showed significant difference in terms of the use of analgesics(OR=2.143),mechanical ventilation(OR=2.071),nasal feeding pathway(OR=1.838),age(OR=1.025),nasal feeding rate(OR=1.042),and nasal feeding volume(OR=1.001).ConclusionApplication of antibiotics,increased age,fasting,oral potassium preparations,increased nasal feeding rate,and increased daily feeding volume are risk factors of EN-related diarrhea;short peptide EN solution,application of analgesics,and increased age are risk factors of EN-related constipation for enteral nutrition patients;and mechanical ventilation,application of nasogastric tube,increased age,increased nasal feeding rate,and increased nasal feeding are risk factors of EN-related relux in enteral nutrition patients.

Purpose To assess the feasibility of measuring maximal density of a region of interest (ROI) with contrast-enhanced CT in quantifying degrees of intestinal ischemia in patients with intestinal obstruction.Materials and Methods Abdominal CT images and reports of 160 patients with intestinal obstruction were retrospectively studied.All the data were reviewed by CT visual evaluation method and measuring maximal density of ROI respectively.The CT visual evaluation took the way of accumulated points,and divided the degrees of intestinal ischemia into five categories.The measuring maximal density of ROI quantified the degrees of bowel enhancement with a bar histogram on CT workstation.The results were compared with the pathological examination.The sensitivity,specificity,positive and negative predictive values and accuracy of the two methods were calculated respectively,and compared.Results The sensitivity,specificity,positive and negative predictive values and accuracy of CT visual evaluation method were 96.7％,72.9％,82.1％,94.4％ and 86.2％,respectively.The sensitivity,specificity,positive and negative predictive values and accuracy of the measuring maximal density of ROI were 68.8％,100.0％,100.0％,71.4％,82.5％,respectively.By measuring the area under the ROC curve,the ROI method (0.995) was more accurate than CT visual evaluation method (0.908) in the diagnosis of bowel ischemia.Conclusion Measuring the maximal density of ROI can quantize bowel wall enhancement.It is a reliable and useful method in the diagnosis of intestinal ischemia,and in accordance with pathological results.

Objective To investigate the effects of erigeron breviscapus (Vant.) Hand-Mazz (erigeron breviscapus) pretreatment on pathology and oxyradical level in the spinal cord after ischemia-reperfusion (I/R) injury in rabbits. Methods A total of 40 New Zealand white rabbits were randomly divided into three groups: sham-operation group with 10 rabbits treated with only abdominal aorta exposure without occlusion, control group with 15 rabbits that underwent ischemia for 50 minutes and treated with matched saline, and experimental group with 15 rabbits that underwent ischemia for 50 minutes and treated with erigeron breviscapus (9mg/kg) injection before ischemia. Malondialdehyde (MDA) level and superoxide dismutase (SOD) activity in the spinal cord were examined at 6 and 24 hours after I/R, respectively. The morphological changes and the number of the spinal cord anterior horn motor neurons were observed and counted under the light microscope and electron microscope, respectively. Results The level of MDA was markedly decreased and SOD activity was increased in the experimental group compared with those in the control group (P<0.01). Compared with that in the control group, the number of motor neurons in the experimental group significantly increased at 24h after I/R (P<0.01) and the morphous of the motor neurons improved. Conclusion Erigeron breviscapus can reduce oxyradical production and the apoptosis of nerve cells, and protect nerve tissue structure and function after spinal cord I/R.

AIM:To observe the effect of captopril on the genesis and development of gastric cancer , and to explore its clinical treatment feasibility for gastric cancer .METHODS:The human gastric cancer cell line AGS was used to establish a tumor model in nude mice , and the model mice were randomly divided into 3 groups: positive control ( 5-fluorouracil) group, normal control (saline) group and experimental (captopril) group.After intraperitoneal injection or intragastric administration of the drugs , the tumor growth curve was determined , and the tumor tissues were also sampled to detect the expression of Ki-67, STAT3, Bax and Bcl-2 by real-time quantitative PCR and immunohistochemistry .The apop-tosis was detected by TUNEL +DAPI staining .RESULTS: The tumor growth curve showed that the tumor model in the nude mice was successfully established .The tumor volumes among groups showed significantly different after 14 d growth. The increase in the tumor volume in normal control group was significantly faster than that in the other two groups , and that in positive control group was the slowest .The expression of Bax in captopril group increased , and the expression of STAT3, Ki-67 and Bcl-2 was reduced as compared with normal control group and positive control group .Compared with normal con-trol group, the apoptotic rate increased significantly , and the protein expression of p-STAT3 and STAT3 decreased obviously in positive control group and captopril group .CONCLUSION:With better feasibility , angiotensin-converting enzyme in-hibitor captopril has a significant effect on treating gastric cancer in the AGS nude mouse model by regulating the expression of STAT3, Bax, Bcl-2 and Ki-67 to accelerate the apoptosis of cancer cells , thus inhibiting tumor growth .

Objective To investigate the protective effects of Bcl-2 associated with athanogene-1 (BAG-1) gene on human neuroblastoma cells (SH-SY5Y) injury induced by hypoxia/reoxygenation,and its influence on heat shock protein 70 (HSP70) expression.Methods The SH-SYSY cells at logarithmic growth phase were collected.Lenti virus mediated RNA interference (RNAi) technology was used to suppress the BAG-1 expression.The cells screened out can be divided into four groups:the cell control group with no lentivirus infection,lentivirus control group (containing only fluorescein protein lentivirus infection),BAG-1 siRNA group (BAG-1 siRNA silencing group),including BAG-1 siRNA-α group and BAG-1 siRNA-β group with lentivirus containing fluorescein protein (GFP) but at different BAG-1 siRNA target sites of silencing.Western Blot was used to detect the protein expression of BAG-1 and HSP70 in target cells after infectious recombination lentivirus for 72 hours;the Cell Counting Kit-8 (CCK-8) was used to detect the activity of four different group cells after hypoxia;the flow cytometry was used to detect the cell apoptosis;the HSP70 mRNA transcription level were detected by real-time fluorescent quantitative reverse transcription-polymerase chain reaction (RT-qPCR) respectively in each group.Results After lentiviral infection for 72 hours,the Western Blot results showed that in the two BAG-1 siRNA silencing groups,the interference effect of BAG-1 siRNA-β group was superior to that of BAG-1 siRNA-α group.The cell viability of each group showed an increase followed by a decrease with the prolongation of hypoxia time,and reaching the peak at 8 hours.After hypoxia for 8 hours being given,the cell viability in BAG-1 siRNA-β group was significantly lower than that of the cell control group,lentivirus control group and BAG-1 siRNA-α group (A value:0.59 ±0.09 vs.0.94±0.12,0.90± 0.11,0.91± 0.14,P ＜ 0.01);the cell apoptosis rate was obviously higher in BAG-1 siRNA-β than that in the above three groups [(34.63 ± 3.46)％ vs.(14.83 ± 3.75)％,(19.93 ± 6.49)％,(16.40± 1.18)％,all P ＜ 0.01].There were no statistically significant differences in the HSP70 protein level and mRNA transcription level between BAG-1 siRNA-3 grroup,and the cell control group,lentivirus control group and BAG-1 siRNA-α group respectively (all P ＞ 0.05).Conclusion BAG-1 gene can play a role in protection of hypoxia nerve cells,reduce the apoptosis,and its protective effect can be independent of HSP70 gene.

Objective: To investigate the mechanisms of mitochondrial apoptosis in spinal cord ischemia-reperfusion injury and the effects of Herba Erigerontis Breviscapi Injection preconditioning intervention. Methods: Sixty Japanese rabbits were divided into sham-operated group, ischemia group, ischemia-reperfusion group (1, 6, 24 and 48 h), and Herba Erigerontis Breviscapi Injection group (1, 6, 24 and 48 h). Clamping the abdominal aorta was used to construct the rabbit model of spinal cord ischemia-reperfusion injury. The rabbits in the ischemia-reperfusion group and the Herba Erigerontis Breviscapi Injection group underwent reperfusion for 1, 6, 24, 48 h respectively after fifty-minute ischemia. The rabbits in the Herba Erigerontis Breviscapi Injection group were administered with Herba Erigerontis Breviscapi Injection at 9 mg/kg 30 minutes before ischemia. Rate of apoptotic cells was measured by flow cytometry; contents of caspase-9 and apoptosis-inducing factor (AIF) in cytoplasm and serum were measured by enzyme-linked immunosorbent assay. Results: Compared with the sham-operated group and the ischemia group, the rates of apoptotic cells, the contents of caspase-9 and AIF in cytoplasm were increased at all time points after reperfusion, and the contents of caspase-9 and AIF in serum were decreased after 1 h and 6 h reperfusion, and increased after 24 h and 48 h reperfusion in the ischemia-reperfusion group. Herba Erigerontis Breviscapi Injection intervention could decrease the rate of apoptotic cells and the contents of caspase-9 and AIF in cytoplasm and serum as compared with those in the ischemia-reperfusion group, and the effects appeared after 1 h reperfusion. Conclusion: The apoptosis of nerve cells after spinal cord ischemia-reperfusion is related to the mitochondrial pathways. Herba Erigerontis Breviscapi Injection can inhibit nerve cell apoptosis by decreasing the contents of caspase-9 and AIF in cytoplasm and serum.

In order to investigate in greater detail the two methods based on Hertz model for analyzing force-distance curve obtained by atomic force microscopy, we acquired the force-distance curves of Hela and MCF-7 cells by atomic force microscopy (AFM) indentation in this study. After the determination of contact point, Young's modulus in different indentation depth were calculated with two analysis methods of "two point" and "slope fitting". The results showed that the Young's modulus of Hela cell was higher than that of MCF-7 cell,which is in accordance with the F-actin distribution of the two types of cell. We found that the Young's modulus of the cells was decreased with increasing indentation depth and the curve trends by "slope fitting". This indicated that the "slope fitting" method could reduce the error caused by the miscalculation of contact point. The purpose of this study was to provide a guidance for researcher to choose an appropriate method for analyzing AFM indentation force-distance curve.
Actins ; Elastic Modulus ; HeLa Cells ; cytology ; Humans ; MCF-7 Cells ; cytology ; Microscopy, Atomic Force