Objectives To investigate the methods of protecting external branch of the superior laryngeal nerve (EBSLN)in carotid endarterectomy and to observe the effect of using these methods in clinical surgery. Methods EBSLN (20 sides)of 10 heads of corpse were studied by using microanatomy from January 2013 to December 2013. The occurrence probability of EBSLN on the lower edge of posterior belly of digastric muscle,medial edge of external carotid artery and upper edge of superior thyroid artery in anatomy triangle was analyzed. The distances from the midpoint of the EBSLN to carotid bifurcation, mandibular angle and mastoid tip were measured. Sixty-five patients with carotid endarterectomy in Tianjin Huanhu Hospital from December 2013 to November 2014 were treated with the protective methods of the relevant EBSLN by using anatomy triangle as a mark. Whether the patients had injury symptoms of EBSLN were followed up after procedure. Results (1)The occurrence probability of 20-side EBSLN in anatomy triangle was 95%(19 sides). The midpoint of EBSLN in the anatomy triangle at the posterior mandibular angle was median 0. 34 (-1. 62 to 2. 43)cm,at the inferior mandibular angle was 1. 28 (-1. 33 to 3. 42) cm,at anterior mastoid tip was 2. 84 (0. 51 to 5. 14)cm,at inferior was 4. 51 (2. 82 to 6. 39)cm,and at anterior superior of the carotid bifurcation was 1. 64 (0. 57 to 3. 78)cm. (2)65 patients who underwent carotid endarterectomy used the protective methods of intraoperative EBSLN. There was no manifestation of EBSLN injury at 3 weeks to 9 months after procedure. Conclusion In carotid endarterectomy,taking an anatomic triangle as a symbol,it is no more than 2 cm of the anterior superior of carotid bifurcation during the separation process. As for the patients with higher or lower position of carotid bifurcation,in the range of crossing rear mandibular angle 0. 50 cm or below the mastoid tip 4. 50 cm for arterial separation should be avoided,and this can effectively protect EBSLN.

Objective To explore the effect of application of hospital-school combined three-dimensional teaching mode in the operation nursing practice teaching.Methods 129 students were collected by cluster sampling,then based on class unit,they were randomized into the experimental group (69 students) and the control group (60 students).The experimental group used hospital-school combined threedimensional teaching mode and the control group used traditional teaching mode.Then,both groups had finished an operation nursing comprehensive theoretical examination and a self-report questionnaire for mastery of basic knowledge of operation nursing.Besides,the experimental group had a questionnaire survey of cognitive evaluation for the new teaching mode.Results The average score of operation nursing comprehensive theoretical examination of the experimental group was (77.81 ±9.87),higher than the control group (70.35±12.37); The self-report score for mastery of basic knowledge of operation nursing of the experimental group was (3.94±0.45),higher than the control group (3.68±0.46).The students of the experimental group also considered that the new teaching mode improved their comprehensive knowledge and innovative abilities,critical thinking ability,humanistic care spirit,communication ability,team cooperation ability and innovation ability,etc.Conclusions The effect of application of hospital-school combined three-dimensional teaching mode in the operation nursing practice teaching was good,but it still had some limitations which needed us to pay attention to and to perfect.

Objective T o observe the changes of cystathionine β-synthase (C B S ) expression in the cere-bral cortex after brain contusion at different tim es. Methods A n experim ental m odel of traum atic brain injury (T B I) in m ice w as established by an im proved w eight-drop device. T hen W estern blotting and im m unohistochem ical exam ination w ere used to detect the C B S expression in cerebral cortex around in-jury at different tim e points (1 h, 6 h, 12 h, 1 d, 2 d, 3 d, 7 d). Results T he results of W estern blotting revealed that the expression level of C B S w as dow n-regulated and reached its low est level at the 3rd days after injury, and then restored to norm al level after 7 days. T he results of im m unohistochem istry show ed that C B S w as present in the norm al brain cortex. C B S expression gradually decreased at the 3rd days after injury, and then restored to norm al level after 7 days. Conclusion C B S has the potential to be a reference index for tim e estim ation after brain contusion in forensic practice.

10.3969/j.issn.2095-4344.2013.25.016

OBJECTIVETo evaluate the efficacy of sacral nerve stimulation (SNS) therapy for fecal incontinence.

METHODSClinical researches which evaluated the efficacy of SNS and were published between 1946 and 2016 were systematically searched from electronic databases, including PubMed, Ovid Medline, Web of Science, Wanfang database and Chinese Journal Full-text Database. Grey area literatures were also searched. Influence of SNS therapy on fecal incontinence episodes (FIE) or Wexner incontinence score (WIS) was systematically evaluated. The statistical analysis was performed by RevMan5.2.

RESULTSA total of 6 studies including 270 patients (147 patients in SNS group and 123 patients in control group) with fecal incontinence were enrolled in this systematic review. SNS therapy was associated with a significant reduction in FIE (SMD=-0.69, 95%CI: -0.97 to -0.41, P<0.001) and a significant reduction in WIS (SMD=-5.05, 95%CI: -8.73 to -1.36, P=0.007). Sensitivity analysis showed that the results of this study were stable and the direction and significance of results were not changed (P=0.000 for both). Publication bias was not found by funnel picture in this study.

CONCLUSIONSNS significantly improves the outcome of patients with fecal incontinence.

Objective: To investigate the level of human blood basic fibroblast growth factor (FGF2) among children with autism spectrum disorder (ASD) and its correlation with behavioral phenotypes, to provide a reference for etiological research of ASD. Methods: ASD Children were selected to get rehabitation training in reseach center of children development behavior in Harbin Medical University and the rehabitation constitution for ASD disabilities in Heilongjiang, 40 children were induded as ASD group, 41 healthy children in Harbin kindergarten was classified as control group. The Autism Behavior Checklist (ABC), Childhood Autism Rating Scale (CARS) and Peabody Picture Vocabulary Test (PPVT) were used to assess the severity and intelligence of ASD children, respectively. Results: No difference was found in FGF2 level between ASD children (4.95 pg/mL) and normal children(8.51 pg/mL)（P>0.05). However, difference in FGF2 level between the two groups were found in 4-year-old group(P<0.05). The level of FGF2 differed across different severity and intelligence of ASD children(P<0.05). Conclusion Abnormal levels of FGF2 in ASD children may correlate with severity of autistic traits and intelligence of children.

Objective:To observe the changes of macular visual function after myopic foveoschisis (MF) and pars plana vitrectomy (PPV) combined with internal limiting membrane peeling and air filling.Methods:A single-center, retrospective study. From October 2018 to October 2019, 29 MF inpatients (32 eyes) in Tianjin Eye Hospital were included in this study. There were 3 males (4 eyes) and 26 females (28 eyes). The age was 63.00±3.45 years old. Equivalent spherical lens degree was -14.16±2.54 D, and axial length was 29.14±1.04 mm. Among them, 3 patients (3 eyes) had lamellar macular holes. All eyes underwent standard pars plana three-channel 25G PPV combined with internal limiting membrane peeling and air filling. Before surgery and at 1, 3, and 6 months after surgery, macular microperimetry was performed with a macular integrity assessment instrument, and the mean retinal sensitivitie (MS) within 10° of the macula, fovea 2° and 4° fixation rates (P1, P2), 63% and 95% bivariate contour ellipse area (BCEA) were recorded. The comparison of MS, P1, P2, 63%BCEA and 95%BCEA at different times before and after surgery was performed by paired t test; the comparison of fixation stability rate was performed by χ 2 test. Results:Compared with before surgery, there were significant differences in the improvement of MS in affected eyes at 1, 3 and 6 months after surgery ( t=-2.208, -3.435,-4.919; P=0.038, 0.002, 0.000). In the pairwise comparison at different times after surgery, only 6 months after surgery and 1 month after surgery were significantly different ( P=0.036). Compared with the preoperative P1, P2, 63%BCEA and 95%BCEA, the P1 and P2 of the eyes gradually increased after surgery, while the 63%BCEA and 95%BCEA gradually decreased, however, the difference was not statistically significant (P1: t=-1.595,-1.698,-1.966; P=0.125, 0.104, 0.062. P2: t=-1.622,-1.654,-1.707; P=0.119, 0.112, 0.102. 63%BCEA: t=1.410, 1.409, 1.553; P=0.172, 0.173, 0.135. 95%BCEA: t=1.412, 1.408, 1.564; P=0.172, 0.173, 0.132). Six months after surgery, all the eyes underwent anatomical repositioning of the macular area, and no serious complications such as full-thickness macular hole and macular hole retinal detachment were found. Conclusions:PPV with internal limiting membrane peeling and air filling is an effective and safe method for MF, and the macular function improved significantly within 6 months postoperatively.

Objective:To investigate the effect of CIRc_0000267 on proliferation, migration, invasion and apoptosis of gastric cancer cells in vitro.Methods:Gastric cancer cell lines with circ_0000267 knockdown and miR-661 overexpression were constructed in vitro, and the expressions of circ_0000267, miR-661 and NGAL mRNA in gastric cancer tissues and cells were detected by qRT-PCR. Cell proliferation, migration, invasion and apoptosis were detected by clonogenesis, Transwell chamber and flow cytometry, and related protein expression was detected by Western blot. Online prediction combined with double luciferase assay and RNA pull down assay was used to verify the targeting relationship between circ_0000267, miR-661 and NGAL. Tumogenesis assay in nude mice was used to observe the effect of circ_0000267 knockout on the growth of gastric cancer cells in vivo. Results:Circ_0000267 was highly expressed in gastric cancer tissues and cells. Knockdown circ_0000267 inhibited the proliferation, migration and invasion of gastric cancer cells and promote apoptosis. Circ_0000267 targeted miR-661, and inhibition of miR-661 partially reversed the effect of circ_0000267 on gastric cancer cells. NGAL was the target gene of miR-661, and overexpression of miR-661 regulated the proliferation, migration, invasion and apoptosis of gastric cancer cells by inhibiting NGAL. Knockdown circ_0000267 targeting miR-661 inhibited NGAL expression in gastric cancer cells; Knockdown circ_0000267 inhibited the growth of gastric cancer cells in vivo.Conclusions:Circ_0000267 may regulate the proliferation, migration, invasion and apoptosis of gastric cancer cells and inhibit the growth of tumors in vivo by regulating the expression of miR-661/NGAL.

Objective:To investigate the effects of serum exosomes of patients with gastric cancer on cisplatin resistance, clonal formation, migration, invasion and apoptosis of the AGS gastric cancer cells, and the corresponding molecular mechanisms.Methods:The experimental study was conducted. The exosomes of patients with gastric cancer was separated from their serum, and the expression of lncRNA HOTTIP was analyzed using the quantitative real time polymerase chain reaction (qRT-PCR). Normal gastric epithelial cell line GES1, gastric cancer cell line AGS and human embryonic kidney cell 293T were cultured in vitro. AGS cells were incubated with exosomes (Exo),with phos-phate buffered saline (PBS) treatment as control, and transfected with si-NC or si-HOTTIP-3, named as Exo group, PBS group, si-NC+Exo group, and si-HOTTIP-3+Exo group. The AGS cells were trans-fected with si-NC, si-HOTTIP-1, si-HOTTIP-2, si-HOTTIP-3, oe-HOTTIP, vector, oe-HOTTIP+miR-138-5p mimic, oe-HOTTIP+mimic NC, miR-138-5p inhibitor, inhibitor NC, miR-138-5p inhibitor+si-TJP1 and miR-138-5p inhibitor+si-NC. They were recorded as si-NC group, si-HOTTIP-1 group, si-HOTTIP-2 group, si-HOTTIP-3 group, oe-HOTTIP group, vector group, oe-HOTTIP+miR-138-5p mimic group, oe-HOTTIP+mimic NC group, miR-138-5p inhibitor group, inhibitor NC group, miR-138-5p inhibitor+si-TJP1 group and miR-138-5p inhibitor+si-NC group. The 293T cells transfected with mimic NC+HOTTIP wt, miR-138-5p mimic+HOTTIP wt, mimic NC+HOTTIP mut, miR-138-5p mimic+HOTTIP mut, mimic NC+TJP1 3'UTR wt, miR-138-5p mimic+TJP1 3'UTR wt, mimic NC+TJP1 3'UTR mut, miR-138-5p mimic+TJP1 3'UTR mut were recorded as the mimic NC+HOTTIP wt group, miR-138-5p mimic+HOTTIP wt group, mimic NC+HOTTIP mut group, miR-138-5p mimic+HOTTIP mut group, mimic NC+TJP1 3'UTR wt group, miR-138-5p mimic+TJP1 3'UTR wt group, mimic NC+TJP1 3'UTR mut group, miR-138-5p mimic+TJP1 3'UTR mut group. The cell counting kit-8 (CCK8) was used to analyze the cisplatin sensitivity of gastric cancer cells. The colony formation experiment was used to analyze the colony formation of gastric cancer cells. The Transwell experiment was used to analyzed cell migration and invasion of gastric cancer cells. The flow cytometry experiment was used to analyze cell apoptosis of gastric cancer cells. The Western bolt assay was used to analyze the expression of exosome marker proteins, including the CD63 and CD81, and the protein of TJP1, the drug-resistance related proteins, including the P-gp and MCL-1. The dual-luciferase assay was used to analyze the targeted relationships among lncRNA HOTTIP, miR-138-5p and TJP1. Observation indicators: (1) expression of lncRNA HOTTIP; (2) resistance of gastric cancer cells to cisplatin regulated by exosome-mediated lncRNA HOTTIP; (3) regulation of cisplatin resistance in gastric cancer cells mediated by miR-138-5p through lncRNA HOTTIP overexpression; (4) targeting of TJP1 gene 3′-untranslated region (UTR) by miR-138-5p; (5) regulation of cisplatin resistance in gastric cancer cells by TJP1 through miR-138-5p inhibition. Measurement data with normal distribution were represented as Mean± SD, and comparison between groups was conducted using the t test. The one-way ANOVA was used for comparison for multiple groups and the Tukey′s test was used for further pairwise compari-son. Count data were described as absolute numbers, and the chi-square test was used for comparison. Correlation analysis was conducted using the Pearson′s test. Results:(1) Expression of lncRNA HOTTIP. The expression of lncRNA HOTTIP in the serum exosome of patients with gastric cancer was higher than that in healthy volunteers, showing a significant difference ( P<0.05). Results of transmi-ssion electron microscopy examination showed that the serum exosomes were circular or oval in shape. Results of Western bolt assay showed the expression of marker proteins of CD63 and CD81 in serum exosomes. (2) Resistance of gastric cancer cells to cisplatin regulated by exosome-mediated lncRNA HOTTIP. Compared with the PBS group, the biochemical half maximal inhibitory concentra-tion (IC50), the number of clone formation, the number of invasive cell, the number of migratory cell, expression of P-gp protein, expression of MCL-1 protein in the Exo group increased, while the cell apoptosis rate decreased, showing significant differences between them ( P<0.05). Compared with the si-NC+Exo group, the IC50, the number of clone formation, the number of invasive cell, the number of migratory cell, expression of P-gp protein, expression of MCL-1 protein in the si-HOTTIP-3+Exo group decreased, while the cell apoptosis rate increased, showing significant differences between them ( P<0.05). (3) Regulation of cisplatin resistance in gastric cancer cells mediated by miR-138-5p through lncRNA HOTTIP overexpression. Compared with the vector group, the IC50, the number of clone formation, the number of invasive cell, the number of migratory cell, expression of P-gp protein, expression of MCL-1 protein in the oe-HOTTIP group increased, while the cell apoptosis rate decreased, showing significant differences between them ( P<0.05). Compared with the oe-HOTTIP+mimic NC group, the IC50, the number of clone formation, the number of invasive cell, the number of migratory cell, expression of P-gp protein, expression of MCL-1 protein in the oe-HOTTIP+miR-138-5p mimic group increased, while the cell apoptosis rate decreased, showing significant differences between them ( P<0.05). (4) Targeting of TJP1 gene 3′-UTR by miR-138-5p. Results of dual-luciferase assay showed that the luciferase activity in 293T cells treatment with mimics of control+vectors of wild type of TJP1 gene 3′-UTR and 293T cells treatment with mimics of miR-138-5p+vectors of wild type of TJP1 gene 3′-UTR was 1.00±0.09 and 0.21±0.03, respectively, showing a significant difference between them ( t=15.02, P<0.05). (5) Regulation of cisplatin resistance in gastric cancer cells by TJP1 through miR-138-5p inhibition. Compared with the inhibitor group, the IC50, the number of clone formation, the number of invasive cell, the number of migratory cell, expression of P-gp protein, expression of MCL-1 protein in the miR-138-5p inhibitor group increased, while the cell apoptosis rate decreased, showing significant differences between them ( P<0.05). Compared with the miR-138-5p inhibitor+si-NC group, the IC50, the number of clone formation, the number of invasive cell, the number of migratory cell, expression of P-gp protein, expression of MCL-1 protein in the miR-138-5p inhibitor+si-TJP1 group decreased, while the cell apoptosis rate increased, showing significant differences between them ( P<0.05). Conclusion:Serum exosomes-mediated lncRNA HOTTIP can promote cisplatin resistance, clonal formation, migration, invasion and apoptosis of gastric cancer cells and inhibit cell apoptosis of gastric cancer cells through regulating the expression of miR-138-5p/TJP1.

Objective To evaluate the effect of transcutaneous electric acupoint stimulation (TEAS)on prognosis in the patients at high risk of postoperative pulmonary complications. Methods Sixty American Society of Anesthesiologists physical statusⅡorⅢpatients of both sexes, aged 65-72 yr, with body mass index of 18-25 kg∕m2, scheduled for elective thoracoscopic and laparoscopic radical resection of e-sophageal cancer, were divided into placebo control group(group C, n=30)and TEAS group(n=30). In group TEAS, bilateral Zusanli(ST36), Hegu(LI4)and Feishu(BL13)acupoints were stimulated for 30 min(disperse-dense waves, frequency 2∕100 Hz, intensity of currents 8-12 mA)starting from 30 min be-fore anesthesia induction.In group C, electrodes were placed on the same acupoints before anesthesia induc-tion, but no current was given.General anesthesia was performed in two groups.Before one-lung ventilation, at 30 min and 2 h of one-lung ventilation and at 1 h after operation(T1-4), blood samples were drawn from the radial artery for blood gas analysis, and oxygenation index and alveolar-arterial oxygen partial pressure difference were calculated.Blood samples were collected from the internal jugular vein at T1-3and 24 h after surgery(T5)for determination of plasma tumor necrosis factor-alpha, interleukin-6(IL-6)and IL-10 con-centrations.The development of postoperative pulmonary complications, time for removal of drainage tube and length of hospital stay were recorded. Results Compared with group C, the oxygenation index at T2-4and plasma IL-10 concentrations at T3were significantly increased, the alveolar-arterial oxygen partial pressure difference at T2,3, plasma tumor necrosis factor-alpha concentrations at T2,3, plasma IL-10 concentrations at T3,5and incidence of postoperative pulmonary complications were decreased, and the time for removal of drainage tube and length of hospital stay were shortened in group TEAS(P<0.05). Conclusion TEAS can improve prognosis in the patients at high risk of postoperative pulmonary complications.