Objective To observe the manifestations and judgement methods of acute rejection reaction of orthotopic liver transplantation in Wistar-SD rats.Methods The survival conditions after operation in rats was observed,and liver function tests and histopathology examination were used to study the manifestations of acute rejection reaction of orthotopic liver transplantation in Wistar-SD rats.Results Wistar-SD experiment group had moderate to severe acute rejection reaction after OLT.Alanine aminotransferase,aspartate aminotransferase,total bilirubin io clood serum after orthotopic liver transplantation in rats was obviously elevated on 1-3 d and 7 d after OLT,which are significantly different from every corresponding time point of control group(P

Objective This study was to detect the expression and level of vascular endothelial growth factor(VEGF)in the acute rejection reaction of orthotopic liver transplantation in rats,which attempted to prove whether VEGF is the key molecule mediating the inter-permeate and inter-enhancement mediated by cells between Angiogenesis and inflammation reaction Methods Expressions of VEGF in plasm in liver and spleen were detected using immunohistochemical staining.The levels of VEGF were measured with enzyme linked immunosorbent assay(ELISA).Results The expression of VEGF in liver,spleen and the level of VEGF in plasma in experiment group were higher than that in the control group(P＜0.05),which were the highest in two days after operation.Conclusion VEGF may play a significant role in the acute rejection reaction of orthotopic liver transplantation in rats.There was a close relation between the expression and level of VEGF in liver,spleen and survival time of graft.VEGF was a kind factor which is expressed in early stage.

Objective To study the relationship between the expression of vascular endothelial growth factor (VEGF) and invasion, metastasis of primary hepatocellular carcinoma (PHCC). Methods Paraffin-embedded specimens from 30 patients with PHCC undergoing radical resection were studied. Agiogenesis was assessed by the expression of VEGF and microvascular density (MVD) using LSAB immunohistochemical staining. Results (1) The level of VEGF and MVD in PHCC was higher than that in the paratumorous tissue (P5*!cm) and small ones (diameter ≤5*!cm). (4) The expression of VEGF was positively correlated with MVD (P

Objective　To study the relationship between the expression of vascular endothelial growth factor (VEGF) and invasion, metastasis of primary hepatocellular carcinoma (PHCC). Methods　Paraffin-embedded specimens from 30 patients with PHCC undergoing radical resection were studied. Agiogenesis was assessed by the expression of VEGF and microvascular density (MVD) using LSAB immunohistochemical staining. Results　(1) The level of VEGF and MVD in　 PHCC was higher than that in the paratumorous tissue (P<0.05). (2) The level of VEGF expression and MVD in PHCC with tumor metastasis or in nonencapsulated tumors were higher than that in PHCC without metastasis or in well encapsulated tumors (P<0.05). 　(3) There was no significant differance in the expression of VEGF and MVD between large PHCC (diameter >5cm) and small ones (diameter ≤5cm). (4) The expression of VEGF was positively correlated with MVD (P<0.01).　　Conclusions　The expression of VEGF and MVD are highly correlated with invasion and metastasis in PHCC, may serve as a parameter for determining tumor metastasis and prognosis which may suggest a new idea to supervise and treat PHCC.

Objective To evaluate the erectile function following transperineal template-guided prostate saturation biopsy (TFPSB).Methods From June 2013 to October 2015,patients underwent prostate biopsy.All patients were indicated for biopsy according to the criteria of "Guidance on diagnosis and treatment of urology in China".Exclusion criteria include medical history of PCa,severe urinary tract infection,severe cardiovascular and cerebrovascular diseases or abnormal blood coagulation.All patients were divided into observation group (TT'PSB) or control group (traditional trans-perineal template-guided prostate biopsy,TTPB) according to patients' condition,pubic anatomy,PSA abnormality,rectal examination,imaging examination and pain tolerance,etc.Patients were evaluated for pre-biopsy erectile function with the international index of erectile function (IIEF-5).All pathology confirmed prostate cancer patients were excluded.Concomitant systemic diseases and medications that would interfere with erectile function were recorded.Patients who withdrew from the trial or used the drugs such 5-phosphodiesterase inhibitors for sexual activity improvement were excluded.Ninety-seven patients in observation group and 84 patients in control group underwent further evaluation with the IIEF-5 questionnaire at 1,3 and 6 months post-biopsy.Results The average age of the observation group and the control group were (64.1 ± 7.9) years and (61.8 ±8.9) years,PSA were (7.2 ± 3.7) ng/ml and (6.7 ± 3.4) ng/ml,prostate volume were (47.8 ±21.5)ml and (49.2 ±22.2) ml,and the BMI were (21.4 ±3.1) kg/m2 and (20.6 ±3.4) kg/m2,respectively.There was no significant difference between the two groups (P ＞ 0.05) in term of above patients' characteristics.The pre-biopsy IIEF-5 score of the observation group and the control group were 19.1 ±4.5 and 19.7 ±4.3,which had no significant difference (t =-0.890,P=0.375).One month after biopsy,the IIEF-5 of two groups were 17.4 ±4.8 and 18.2 ±4.5 respectively and both group had statistically significant difference when compared with pre-biopsy (both P ＜ 0.05),however,there was no statistical significance at 3 and 6 months after biopsy.Besides,no significant difference of the IIEF-5 score was detected between two groups in 1,3 and 6 months.Conclusions Although TTPSB may resulted in temporary (1 month) post-biopsy erection dysfunction,the erectile function recovered to pre-biopsy level at 3-6 months post-biopsy.TTPSB did not increase the risk of ED compared with traditional TTPB.

Objective: To evaluate the effect of multimodal analgesia using periprostatic nerve block anesthesia (PNB) combined with flurbiprofen in patients undergoing transperineal template-guided prostate biopsy (TTPB). Methods: Totally 166 patients (aged (68.2±9.1) years, range: 47 to 81 years) who received TTPB from October 2017 to June 2018 at Department of Urology, Northern Jiangsu People′s Hospital Affiliated to Yangzhou University were enrolled prospectively. All the patients were randomly divided into 2 groups. The observation group (n=79) was given flurbiprofen axetil 1 mg/kg intravenously for half an hour before operation and lidocaine was used for PNB before the biopsy. The control group (n=87) was given normal saline combined with PNB. A visual analog scale (VAS) and visual numeric scale (VNS) were used to assess the patients′ pain and quantify their satisfaction at two time points: VAS-1 and VNS-1: during biopsy procedure, VAS-2 and VNS-2: 30 min after the procedure. The date were compared by t test, χ² test, Fisher exact test and two-way repeated measures anova analysis between the 2 groups. Results: The age, total prostate volume, serum prostate-specific antigen and the number of cores were comparable among the 2 groups (P>0.05). The VAS-1 scores of the control group and the observation group were 2.8±1.7, 1.9±1.2, respectively, and the VNS-1 were 3.1±0.7, 3.4±0.3, respectively. The VAS-1 were significantly lower in observation group than in control group (F=3.904, P=0.000). Conversely, the VNS-1 were higher in observation group (F=3.526, P=0.000). At 30-minute postoperative, the VAS-2 and VNS-2 were 0.7±0.4 and 3.7±0.2 in the control group, respectively. The VAS-2 and VNS-2 were 0.6±0.5 and 3.8±0.1 in the observation group, respectively. There were no significant differences in the pain scores or the satisfaction scores between the 2 groups (F=1.429, 2.825; P=0.136, 0.083). The incidence of overall complications was 26.4% (23/87) in the control group and 25.3% (20/79) in the observation group, with no statistical difference between the 2 groups (χ²=0.027, P=0.869). And the complications had no statistically significant difference among the 2 groups including hematuria, urinary retention, infection, hematospermia, vascular and neurological reactions, nausea, vomiting, dizziness, headache, and respiratory depression (P>0.05). Conclusion The multimodal analgesia induced by PNB and flurbiprofen could effectively relieve the pain for patients who received TTPB.

Objective: To investigate the clinical value of modified transperineal template-guided prostate biopsy (mTTPB) in the detection of prostate cancer. Methods: A total of 217 patients were enrolled in this study. All the patients were randomly divided into 2 groups. The control group (n=112) underwent transperineal template-guided prostate biopsy (TTPB) which was traditional transperineal template-guided 11-region biopsy. On the basis of the control group, the apex of prostate was divided into four areas for biopsy in the observation group (mTTPB). The positive rate of apex and the incidence of complications were analyzed. The prostatic specimens from the radical prostatectomy underwent whole mount sections examination. The prostate biopsy results were compared with the postoperative pathological results. Results: The average age of the control group and the observation group were (68.5±7.9) years and (67.3±8.5) years, PSA were (31.2±18.9) ng/ml and (29.7±19.5) ng/ml, prostate volume were (44.6±15.2) ml and (41.3±17.3) ml, respectively. In the control group, the positive rates of prostate cancer in 1-10 region were 24.1% (27/112), 27.7%(31/112), 23.2% (26/112), 28.6% (32/112), 26.8% (30/112), 25.0% (28/112), 26.8% (30/112), 19.6% (22/112), 25.9% (29/112), 25.0% (28/112), respectively, with an average of 25.3%. In the observation group, the positive rates in 1-10 region were 27.6% (29/105), 28.6% (30/105), 22.9% (24/105), 26.7% (28/105), 25.7% (27/105), 24.8% (26/105), 27.6% (29/105), 21.9% (23/105), 27.6% (29/105), 26.7% (28/105), respectively, with an average of 26.0%. There was no statistical difference between the two groups (P=0.904). The positive rate of apical prostate cancer in the control group and observation group was 37.5% (42/112) and 44.8% (47/105), respectively, and there was no statistical difference between the two groups (P=0.277). Patients were grouped according to PSA>20 ng/ml and PSA≤20 ng/ml. When PSA>20 ng/ml, the positive rate of apex was 58.6% (34/58) and 56.6% (30/53) respectively in the control group and the observation group, and there was no statistical difference between the two groups (P=0.830). When PSA≤20 ng/ml, the positive rate of apex was 14.8% (8/54) in the control group and 32.7% (17/52) in the observation group, with statistically significant differences (P=0.030). Before radical prostatectomy, 12 cases (57.1%) in the control group and 19 cases (73.1%) in the observation group showed apical invasion by biopsy. Results of whole mount sections examination in the control group showed that there were 19 cases (90.5%) with apical invasion, which was statistically different from that before surgery (P=0.035). The results of whole mount sections examination in the observation group showed that there were 23 cases (88.5%) with apex invasion, which had no statistical difference compared with that before surgery (P=0.291). There were no significant differences in the incidence of hematuria, fever, urinary retention and perineal discomfort between the observation group and the control group (all P>0.05). Conclusions mTTPB can significantly improve the detection rate of apical prostate cancer without increasing the incidence of complications, especially for patients with PSA≤20 ng/ml. Hence is safe and efficacious.

Objective:To identify a novel bombesin bioactive peptide from the skin secretion of Hylarana Latouchii, and to explore its effect on insulin secretion in islet cells.Methods:The skin secretion from Hylarana Latouchii was extracted by electrical stimulation, and the single chain of bombesin peptide was cloned and sequenced. The peptide QUB2995 was synthesized via solid-phase synthesis, then purified using reversed-phase high performance liquid chromatography (HPLC). Matrix assisted laser desorption time-of-flight mass spectrometry (MALDI-TOF) was applied to validate. QPCR and ELISA were used to probe the effect of QUB2995 on insulin secretion in MIN6 and INS-1 cells.Results:A novel bombesin peptide named QUB2995 (GAFGDFLKGAAKA GALKILSIAQCKLSGTC) was found in the skin secretion of Hylarana Latouchii through molecular cloning. The bioactive peptide could significantly promote the proliferation and insulin secretion from mouse islet MIN6 cells and rat islet INS-1 cells. The effect reached a climax at the concentration of 10 -5 mol/L. Conclusion:A novel bombesin bioactive peptide named QUB2995 was found from Hylarana Latouchii. It could significantly promote insulin secretion in MIN6 cells of mouse islets and INS-1 cells of rat islets, indicating its potential in the treatment of diabetes.

BACKGROUND:Frog active peptides have rich activities,such as antibacterial and anti-tumor,and are expected to solve the problem of antibiotic resistance. OBJECTIVE:The active peptide QUB2984 was discovered in the skin secretions of Agalychnis callidryas.Its structure and properties were simulated by bioinformatics.The peptide was synthesized,purified,and identified and its biological functions were investigated. METHODS:Agalychnis callidryas skin secretions were collected by electrostimulation.The sequence of QUB2984 was obtained through constructing a cDNA library with isolated mRNA.BLAST was used for peptide sequence alignment.Besides that,Iterative Threading ASSEmbly Refinement(I-TASSER)and HeliQuest tools were used for protein secondary structure simulation.It was synthesized by solid-phase peptide synthesis,purified by reverse-phase high-performance liquid chromatography,and structurally confirmed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry.The purified peptide was used to evaluate its biological activity.Its antibacterial effect was evaluated by the minimum inhibitory concentration method.Its cytotoxic effect was detected by MTT assay.Its safety was investigated by a hemolysis test. RESULTS AND CONCLUSION:(1)Peptide QUB2984 had basically α-spiral structure,with a relatively intact hydrophobic surface,and a certain destructive ability to biofilm.The third amino acid position of QUB2984 was composed of W and had a G-X-G structure.(2)The minimum inhibitory concentration of QUB2984 against gram-positive Staphylococcus aureus was 2 μmol/L,the minimum inhibitory concentration against gram-negative Escherichia coli was 2 μmol/L,and the minimum inhibitory concentration against the fungus Candida albicans was 8 μmol/L.(3)The active peptide QUB2984 had obvious inhibitory effect on human non-small cell lung cancer cells NCI-H838 at 10-5 mol/L concentration,and the hemolytic effect on horse red cells at 64 μmol/L concentration was 50%.(4)The results showed that QUB2984 had anti-bacterial and anti-cancer activity,and it had a positive charge of +3,which was conducive to contact with bacteria or cells.

Stroke， including ischemic stroke and hemorrhagic stroke， has the characteristics of high morbidity， high disability rate， high mortality rate， high recurrence rate， and high economic burden. Ischemic stroke is the most common type of stroke， which is mainly caused by intracranial artery occlusion. The clinical manifestations of patients are hemiplegia， aphasia， sensory disturbance， and other neurological deficits， accompanied by gastrointestinal symptoms such as constipation and gastrointestinal bleeding. Intestinal flora plays an important role in the pathogenesis of ischemic stroke， and its potential biological effects have received extensive attention. Intestinal flora can not only affect intestinal barrier function but also regulate gastrointestinal immunity and affect host homeostasis. In recent years， traditional Chinese medicine （TCM） has shown remarkable effects and small adverse reactions in the prevention and treatment of cerebral ischemia. The research on the effect of TCM in improving cerebral ischemia injury by regulating the structure and metabolism of intestinal flora and maintaining the function of intestinal flora has gradually become a hot topic. Based on the interaction between TCM and intestinal flora in relevant literature in recent years， this review investigated the mechanism of anti-cerebral ischemic injury of TCM via regulating intestinal flora structure， affecting intestinal flora metabolism， and regulating body immunity and made a summary， proving a basis for further elucidating the role of intestinal flora in cerebral ischemia and the mechanism of TCM in prevention and treatment of ischemic stroke.