Objective To investigate the optimal digestion method for the preparation of single-cell suspensions from mouse small intestinal lamina propria.Methods Ten mouse small intestines of uniform length were collected and randomly divided into two groups.Each group was used to prepare lamina propria single-cell suspensions by enzymatic digestion with collagenase A or collagenase Ⅷ.We compared the effects of these two enzymatic digestion method on the cell yield,cell activity,and cell surface antigens of the single-cell suspensions.Single-cell suspensions prepared by the superior enzymatic digestion method were then subjected to flow cytometry assay.Results Compared to collagenase-Ⅷ-based enzymatic digestion,collagenase-A-based digestion result ed in a higher cell yield(9.48±1.10)× 109vs(4.18±1.02)×109and higher proportions of live cells(86.36±3.32)％vs(61.62±10.93)％,active CD45+cells(57.19±5.11)％vs(26.01±11.44)％,active CD3+cells(8.73±2.89)％vs(4.52±2.49)％,active CD4+cells(6.19±2.09)％vs(3.22±1.91)％,and active B220+cells(15.06±4.27)％vs(5.07±2.20)％,providing high-quality cells for subsequent flow assays.Conclusions The collagenase A-based enzymatic digestion method is more suitable for the preparation of ingle-cell suspensions from the lamina propria of ouse small intestines.

Objective To explore the application effect of healthcare failure mode and effect analysis (HFMEA) in nursing risk management of emergency surgery for patients with traumatic brain injury (TBI). Methods Taking the HFMEA model as the theoretical framework, the emergency surgical process of TBI was analyzed, a flow chart was drawn, and the potential risk factors were analyzed. Corresponding intervention programs were adopted for high-risk failure modes, and 80 patients from November 2022 to April 2023 were conducted with routine management before implementation of HFMEA (control group), the other 80 patients from May to October 2023 were conducted with management after implementation of HFMEA (observation group). The surgical efficiency (preoperative waiting time, surgical preparation time, anesthesia induction time, postoperative handover time), the incidence rates of adverse events in the operation room (incomplete preparation of supplies, contamination of surgical drapes, insufficient or defective intraoperative instruments, errors in writing nursing records, incorrect or missing items in handover information), the incidence rates of perioperative complications (low intraoperative body temperature, pressure injury, abnormal fluctuations in vital signs during surgery, wound infection, lung infection) and satisfaction degree of surgical doctors to nurses for current operations (itinerant nurses, instrument nurses) were compared between two groups. Results The observation group had significantly shorter preoperative waiting time, surgical preparation time, anesthesia induction time, and postoperative handover time compared to the control group (P < 0.01). The incidence rate of adverse events in the operation room for the observation group was 16.25%, which was significantly lower than 46.25% of the control group (P < 0.01). The incidence rate of complications was 8.75% in the observation group, which was significantly lower than 26.25% in the control group (P < 0.01). Satisfaction degree of the doctors to nurses for the current operation in the observation group was significantly higher than that in the control group (P < 0.05). Conclusion Application of the HFMEA model in the perioperative nursing management of patients with emergency surgery for TBI can effectively improve surgical efficiency, reduce the incidence rates of nursing-related adverse events and complications, and enhance satisfaction degree of doctors to nurses in operation room.

Liver is the central hub regulating energy metabolism during feeding-fasting transition. Evidence suggests that fasting and refeeding induce dynamic changes in liver size, but the underlying mechanisms remain unclear. Yes-associated protein (YAP) is a key regulator of organ size. This study aims to explore the role of YAP in fasting- and refeeding-induced changes in liver size. Here, fasting significantly reduced liver size, which was recovered to the normal level after refeeding. Moreover, hepatocyte size was decreased and hepatocyte proliferation was inhibited after fasting. Conversely, refeeding promoted hepatocyte enlargement and proliferation compared to fasted state. Mechanistically, fasting or refeeding regulated the expression of YAP and its downstream targets, as well as the proliferation-related protein cyclin D1 (CCND1). Furthermore, fasting significantly reduced the liver size in AAV-control mice, which was mitigated in AAV Yap (5SA) mice. Yap overexpression also prevented the effect of fasting on hepatocyte size and proliferation. Besides, the recovery of liver size after refeeding was delayed in AAV Yap shRNA mice. Yap knockdown attenuated refeeding-induced hepatocyte enlargement and proliferation. In summary, this study demonstrated that YAP plays an important role in dynamic changes of liver size during fasting-refeeding transition, which provides new evidence for YAP in regulating liver size under energy stress.

ObjectiveTo investigate whether the quantitative magnetic resonance imaging （MRI） measurement of pancreatic fat content may indicate impaired glucose tolerance （IGT） or type 2 diabetes mellitus （T2DM）. MethodsA total of 108 subjects （43 men， 65 women； aged 47.9±12.1 years） were enrolled in this study， including 27 with T2DM， 29 with IGT and 53 with normal glucose tolerant （NGT）. NGT individuals were subdivided into younger group （<40 years old） and elder group （≥40 years old）. All the participants underwent standard laboratory tests for insulin resistance and β-cell dysfunction. MRI scanning with the iterative decomposition of water and fat with echo asymmetry and least square estimation image quantification （IDEAL-IQ） technique was used to determine fat distribution in the pancreas. The correlation between pancreatic fat volume fractions （PFVF） and laboratory parameters was analyzed. The feasibility of using PFVF to predict IGT and T2DM was evaluated. ResultsPFVF was significantly higher in T2DM patients than that in both IGT and NGT patients， with lowest PFVF in NGT patients. Logistic regression indicated that PFVF was an independent risk factor of glucose abnormality （IGT and T2DM）. ROC curve analysis showed that the cut-off value for PFVF to predict glucose abnormality （IGT and T2DM） was 5.68% （P< 0.001； AUC： 0.871； sensitivity： 92.7%； specificity： 71.7%）. ConclusionMRI fat quantitative assay can quantitatively determine fat content in the pancreas， providing a non-invasive biomarker for the prediction of glucose abnormality （IGT and T2DM）.

Objective:To explore the effective method for treatment of small and short penis after hypospadias surgery.Method:s From November 2017 to November 2018, 57 children aged 4 to 14[mean age(7.91±2.89)years] with hypospadias who met the diagnostic criteria of small penis were reexamined at the First Affiliated Hospital of Xin-xiang Medical University.They were randomly divided into the physical treatment group and the drug treatment group according to the order of visits, and the untreated patients were included in the control group.Among them, 21 patients in the physical treatment group were treated with penile rehabilitation therapy apparatus, supplemented by Salvia mil-tiorrhiza bath (30 minutes/time, once/day, 10 days), and 20 patients in the drug treatment group were treated with Testosterone cream topically (3 times/day, 10 days). Penile relaxation length, stretch length, transverse and longitudinal diameters of glans in 2 groups before and after the treatment were measured.The relevant indexes of 16 patients in the control group measured before and after 10 days and compared with those in the treatment group.Result:s (1)The penile relaxation length in the physical treatment group increased from (25.48±6.13) mm to (30.72±6.49) mm, the length of stretch increased from (34.90±7.71) mm to (41.08±8.43) mm, the transverse diameter of glans increased from (14.81±3.40) mm to (16.57±3.42) mm, and the longitudinal diameter increased from (13.94±3.15) mm to (15.82±3.52) mm, and the differences were statistically significant (all P<0.05). (2)The penile relaxation length in the drug treatment group increased from (21.07±4.26) mm to (31.32±4.72) mm, the length of stretch increased from (31.94±7.96) mm to (45.39±7.24) mm, the transverse diameter of glans increased from (13.38±1.77) mm to (16.64±2.10) mm, and the longitudinal diameter increased from (13.09±1.77) mm to (16.62±1.86) mm, and the differences were statistically significant (all P<0.05). (3)There was no significant difference in penile relaxation length, the length of stretch, transverse diameter and longitudinal diameters of glans before and after 10 days in the control group (all P>0.05). (4) Compared with the control group, the penile relaxation length, the length of stretch, transverse diameter and longitudinal diameters of glans in the physical treatment group increased significantly, and the differences of growth values between the two groups were statistically significant (all P<0.05). (5) Compared with the control group, the penile relaxation length, the length of stretch, transverse diameter and longitudinal diameters of glans in the drug treatment group also increased significantly, and the difference of growth values between the two groups were statistically significant (all P<0.05). (6) The growth of penile relaxation length, the length of stretch and transverse and longitudinal diameters in the drug treatment group were higher than those in the physical treatment group, and the difference of growth values were statistically significant (all P<0.05). Conclusions:Both the negative pressure suction method and topical application of Testosterone cream are effective in the treatment of small and short penis after hypospadias surgery.However, Testosterone cream is difficult to obtain, and the treatment of negative pressure suction is simple, noninvasive, painless and free of adverse reactions.

OBJECTIVE:To evaluate the effectiveness and safety of repaglinide combined with metformin versus glimepiride combined with metformin in the treatment of type 2 diabetes mellitus (T2DM),and to provide evidence-based reference for the clinic. METHODS:Retrieved from CJFD, VIP, Wanfang database, PubMed, Embase, Medline and Cochrane Library, randomized controlled trials (RCTs) about therapeutic efficacy (HbA1c, FPG, 2 hPG) and safety (the incidence of ADR,hypoglycemia and gastrointestinal reaction)of repaglinide combined with metformin(trial group)versus glimepiride combined with metformin(control group)in the treatment of T2DM were collected. Meta-analysis was performed by using Rev Man 5.2 statistical software after data extraction and quality evaluation with Cochrane systematic evaluation manual. RESULTS:A total of 12 RCTs were included,involving 957 patients. Results of Meta-analysis showed that the decrease of 2 hPG in trial group was significantly better than control group,with statistical significance [MD=-0.70,95％CI(-1.02,-0.38),P<0.001]. There was no statistical significance in the decrease of HbA1c [MD=0.00,95％CI(-0.24,0.25),P=0.98] or FPG [MD=0.10,95％CI(-0.17,0.36),P=0.47],the incidence of ADR [OR=0.54,95％CI(0.28,1.06),P=0.07],hypoglycemia [OR=0.52,95％CI(0.13,2.06),P=0.35] or gastrointestinal reactions [OR=0.60,95％CI(0.15,2.41),P=0.47] between 2 groups. CONCLUSIONS:Repaglinide combined with metformin is better than glimepiride combined with metformin in reducing 2 hPG,but both of them have similar safety.

Objective To investigate the effect of chewing gum on intestinal function in patients with colorectal.Methods From October 2015 to April 2016, 69 patients with colorectal cancer radical operation treated in our emergency surgical ward and gastrointestinal surgical ward were selected as the research object. They were randomly divided into control group (n=32) and observation group (n=37). The patients in the control group received standard care while the patients in the observation group received standard care adding chewing gum and then observed all the patient's recovery time of anal (stoma) exhaust time, defecation time, hospitalization time, hospitalization expenses and complications.Results The data of anal(stoma) exhaust time (45.91±16.36)hour, hospital stay (11.13±2.65) day of observation group were significantly lower than those of control group time (t=2.52, 2.63,P<0.05). Comparing the complications after treatment of patients in two group, there were 19 patients suffering abdominal distension and 7 patients suffering nausea and vomit in the observation group, they were lower than the control group with significant differences (χ2=5.12, 4.38;P<0.05). But there were 4 patients with stomach tube in observation group could not tolerate chewing gum.Conclusions Chewing gum can effectively shorten the recovery time of gastrointestinal function of colorectal cancer patients after radical operation and hospital stay. Meanwhile, it can reduce the incidence of abdominal distension, nausea and vomiting.

Objective To study the effect of hepatitis E virus Open Reading Frame 3 protein on the proliferation and cell cycle of Chang liver cells (CCL13).Methods The pDsRed-Monomer-N1-ORF3 recombinant plasmid was transfected into CCL13 cells via lipofectamine 2 000 reagent,then assayed cell proliferation by MTT assay;Flow cytometric was used to detect cell cycle after dying with PI;and the cell cycle proteins expression were detected by western-blotting.Results pDsRed-Monomer-N1-ORF3 recombinant plasmid was successfully transfected into CCL13 cells,Expression of ORF3 can inhibit the proliferation activity of CCL13 cells from the 3rd day (P ＜ 0.01),and prevent the cell cycle in G0/G1 phase;Western Blot showed expression of cell cycle proteins cyclin D1 and cyclin E1 were decreased in ORF3 group compared with the control group;However,the expression of cyclin A2 and cyclin B1 had no significantly difference in three groups.Conclusions HEV-ORF3 can inhibits the proliferation activity of CCL13 cells by inhibition expression of cell cycle proteins cyclin D1 and cyclin E1.

Objective:To observe the efficacy of calcined bovine bone(CBB)in the repair of animal bone defects.Methods:Ca-nine alveolar bone defect model,rat and rabbit calvarial critical size defect models were established respectively.All animals were randomly divided into 2 groups(n=6).The defects in the experimental group were repaire with CBB,those in the control group were not treated.The effects were observed by HE staining,Micro CT and Masson trichrome staining.Results:Canin alveolar bone de-fects were well repaired 8 week after operation.The skull defects in rabbits and SD rats were replaced by new bone 1 2 and 8 weeks after operation respectively.In all the control groups bone defects were not healed or not completely repaired.Conclusion:CBB is ef-fective in the repair of bone defects.

Objective To explore the clinical effect and lung CT change of long-term used of low-dose roxithromycin in treatment for bronchial expansion patients in stable phase. Methods 94 cases collected in the Department of Respiration, The Second Hospital Affiliated to Suzhou University from February 2011 to December 2012 were diagnosed as bronchiectasis, 34 cases in control group were given oral treatment for ambroxol 30 mg, three times one day, 60 cases in treatment group were added roxithromycin 75 mg on basis of control group, two times one day. Patients in two groups were both treated for 6 months. The therapeutic effect and the score of life quality and dyspnea scores in two groups were observed, and the changes of CT data were compared before and after treatment. Results After treatment, the life quality score and dyspnea score of two groups were all improved, but the treatment group was signiifcantly higher than the control group (P<0.05). The effective rate in treatment group was 86.67%, which was signiifcantly higher than 70.59%in the control group (P<0.05). After treatment, chest CT imaging score of patients in treatment group were improved, signiifcantly better than that in the control group (P<0.05). Conclusion Long-term low dose administration of roxithromycin can control and stable bronchiectasis symptoms, and improve signs and symptoms .