A strain of canine parvovirus (CPV) was isolated from feces of an ill puppy in an animal hospital in Wuhan, China. It was designated as CPV/WH02/06. This isolate was identified as serotype CPV-2a by the hemagglutination test, CPV Ag detection strip, electron microscopy, and PCR. The vp2 gene was cloned and sequenced and assigned GenBank accession number EU377537. A 1242 bp segment of the 5' region of the vp2 gene was cloned and inserted into the binary vector pBI121 and used for Agrobacterium-mediated tobacco transformation. Transgenic tobacco plants were selected on MS medium supplemented with 100 μg/mL kanamycin and 100 μg/mL timentin. Integration of the vp2 gene into the tobacco genome was confirmed by PCR using T1 progeny plants, and the expression of the VP2 protein was confirmed by Western blotting.

Objective To summarize our experiences in the diagnosis and treatment of giant colonic lipoma,and enhance awareness of the disease and avoid misdiagnosis.Methods We retrospectively analyzed the clinical data of 5 cases of giant colonic lipoma that were mistaken for colon cancer,and evaluate the possible reasons of misdiagnosis.Results Colonic lipoma is an uncommon benign tumor.Most cases with larger tumor size were misdiagnosed as colon cancer when they suffered from the symptoms of abdominal pain,bloody stool,abdominal mass,even bowel obstruction and intussusception.Barium enema and colonoscopy are important diagnostic means.Contrast enhanced CT scan can provide definitive topographical information identifying a colonic lipoma.Surgical resection was performed in all the 5 cases.Conclusion Giant colonic lipoma with intussusception can mimic a colonic carcinoma leading to misdiagnosis.

Objective To investigate the expression of monocyte chemoattractant protein-1(MCP-1)and CCR2 in peripheral blood and pleural fluid of tuberculous pleurisy patients.Methods Flow cytometry was used to detect the expression of CCR2 in the peripheral blood mononuclear cell and pleural fluid cell of tuberculous pleurisy patients;ELISA was used to detect the content of MCP-1 in serum and pleural fluid.Results MCP-1 in surem and pleural fluid and CCR2 in the peripheral blood mononuclear cell and pleural fluid cell of tuberculous pleurisy patients was significantly higher than that of the healthy normal controls[MCP-1:(340.8±220.8)and(9.0±3.8)ng/L,P＜0.01;CCR2(18.2±10.1)%and(6.9±3.5)%,P＜0.05];Both MCP-1 and CCR2 were detected in pleural fluid and both of them were positivley correlated(r=0.227,P＜0.05).Conclusion The expression of MCP-1 and CCR2 in peripheral blood of tuberculous pleurisy patient are significantly elevated which are significance molecule participating in the pathogenesis of tuberculous pleurisy.

BACKGROUND:The metastatic potential of hepatocelular carcinoma cels is key factor influencing patient’s prognosis. To observe the effect of human bone marrow mesenchymal stem cels on metastasis of hepatocelular carcinoma is of great significance for improving the lifetime of hepatocelular carcinoma patients. OBJECTIVE:To explore the biological effect of human bone marrow mesenchymal stem cels on hepatocelular carcinoma cels with different metastatic potentials. METHODS:Human bone marrow mesenchymal stem cels and hepatocelular carcinoma cel suspension with high and low metastatic potentials were respectively injected into the Transwel chamber, and after 36 hours of co-culture, ELISA method was used to detect the absorbance value as wel as cel counting method was used to observe the changes in the invasion ability of hepatocelular carcinoma cels. The effects of human bone marrow mesenchymal stem cels on the proliferation of hepatocelular carcinoma cel suspension with high and low metastatic potentials were determined using cel counting kit-8. PCR method was adopted to measure the expression of osteopontin, bone specific sialoproteins, integration (alpha V), transforming growth factor beta 1 and programmed cel death protein 5. RESULTS AND CONCLUSION:(1) The number of migrated hepatocelular carcinoma cels was significantly lower in the co-culture group than the single culture group, and based on the semi-quantitative detection of invasion ability, the absorbance value of the co-culture group was significantly lower than that in the single culture group (P < 0.05). (2) The expression of osteopontin and bone specific sialoproteins was significantly decreased in the co-culture group with high metastatic potential (P < 0.05), but there was no change in the expression of integration (alpha V) (P> 0.05). In the co-culture group with low metastatic potential, the expression of osteopontin, bone specific sialoproteins, and integration (alpha V) were declined remarkably (P < 0.05). (3) Results from the semi-quantitative detection of proliferation ability showed that the absorbance value of the co-culture group was significantly higher than that of the single culture group (P < 0.05). (4) In the co-culture group with high metastatic potential, the expression of transforming growth factor beta 1 was up-regulated significantly (P< 0.05), but the expression of programmed cel death protein 5 showed no changes (P > 0.05). However, in the co-culture group with low metastatic potential, the expression of transforming growth factor beta 1 and programmed cel death protein 5 was both increased dramaticaly (P < 0.05). These findings suggest that the human bone marrow mesenchymal stem cels reduce the invasion ability of hepatocelular carcinoma cels, and enhance their ability of proliferation.

OBJECTIVETo study the anatomy basis for the clinical application of the adjacent horn shaped perforator fasciocutaneous flap for the reconstruction of small and medium-sized defects in the trunk area.

METHODS(1) Ten adult antiseptic cadavers (20 sides) were perfused with red latex. The skin blood supply, line of the blood vessels, branches in accordance with the distribution and crossing were observed. (2) Fifteen cases with defects in the trunk were treated with the adjacent horn shaped perforator fasciocutaneous flaps. The defects size ranged from 5 cm x 5 cm to 13 cm x 13 cm with the size of the flaps ranging from 10 cm x 6 cm to 35 cm x 15 cm.

RESULTSThe trunk skin is supplied by mainly 17 groups arteries such as thyrocervical trunk, internal thoracic artery, posterior intercostal arteries, superior epigastric artery, arteria epigastrica inferior, lumbar arteries, and so on. The perforators (diameter > 0.5 mm) numbers are about 20, 40, 24, 6, on the chest, abdomen and perineum, upper back, waist, respectively. All the flaps survived completely with primary healing both on donor and recipient sites. The flaps color, texture, function and appearance were satisfactory during the follow-up period of 1-24 months.

CONCLUSIONSThe adjacent horn shaped perforator fasciocutaneous flap should be designed flexibly. The defects in the donor sites could be closed directly without skin graft. It is an effective, easy and ideal method for the reconstruction of large defects in the trunk.

Adolescent ; Adult ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Perforator Flap ; Skin Transplantation ; Torso ; surgery ; Young Adult

OBJECTIVETo explore the anatomic basis and clinical application of the horn shaped perforator flap pedicled with the angular artery for the reconstruction of midface defect.

METHODS(1) 10 fresh cadavers were perfused with a modified guiding oxide gelatin mixture for three-dimensional visualization reconstruction using a 16-slice spiral computed tomography scanner and specialized software (Materiaise' s interactive medical image control system, MIMICS). The origin and distribution of the angular artery perforator were observed. (2) Between July 2012 and July 2014, twenty-one patients underwent operations for the reconstruction of midface defect. Ten patients had squamous cell carcinoma, nine patients had basal cell carcinoma and two patients had nevus. The flaps' size ranged from 1.5 cm x 3.5 cm to 2.5 cm x 5.0 cm.

RESULTSThe facial artery branches the lateral nasal artery 1 cm from the outside corner of the mouth, subsequently strenches to inner canthus continuing as the angular artery. The angular artery anastomoses extensively with the dorsal nasal artery and the infraorbital artery. All the flaps survived. The patients were satisfied with the final aesthetic and functional results.

CONCLUSIONSThe flap can be designed flexibly and simply with reliable blood supply. The donor sites could be closed directly without skin graft, it is a simple and fast method for the reconstruction of midface defect.

Anastomosis, Surgical ; methods ; Arteries ; anatomy & histology ; Cadaver ; Carcinoma, Basal Cell ; surgery ; Carcinoma, Squamous Cell ; surgery ; Face ; blood supply ; Facial Neoplasms ; surgery ; Humans ; Nevus ; surgery ; Nose ; blood supply ; Perforator Flap ; blood supply ; transplantation ; Skin Neoplasms ; surgery ; Skin Transplantation ; Software ; Tomography, Spiral Computed

Objective To evaluate the expression of CD47 and calreticulin and their relationships with clini-copathologic features,prognosis in infiltrating ductal breast carcinomas.Methods The expression of CD47 and calret-iculin was evaluated in 128 cases with infiltrating ductal breast carcinomas by means of immunohistochemistry,and the relationship between CD47 and other clinicopathologic factors as well as prognosis were analyzed.Results The expression level of CD47 in cancer nest was 81/128,which was lower than 106/128 in its adjacent normal epithelium (χ2 =12.400,P<0.05),but calreticulin expression was 50/128 and was higher in cancer nest(χ2 =21.510,P<0.05 ).In addition,the expression level of calreticulin had certain difference in different molecular subtype (χ2 =21.510,P<0.05),and an increased expression in Basal-like and Her-(2 +)/ER(-)subtypes.The expression level of CD47 was associated with tumor sizes,histological tumor grade and lymph node involvement(χ2 =11.400, 4.732,5.432,all P<0.05),and the expression level of calreticulin was associated with histological tumor grade and distant metastasis(χ2 =10.810,6.770,all P<0.05).The 5 -year survival rate of patients performed following-up was 68.37%.Univariate analysis indicated the survival rate in 5 years of high level CD47,calreticulin expression was higher than that of low level CD47,calreticulin expression(χ2 =5.231,13.069,all P<0.05).Conclusion The expression of CD47,calreticulin is associated with index of dysprognosis and survival rate in 5 years,so combined CD47 and calreticulin detect may be of prognostic value in patients with breast caner.

Objective To evaluate the effects of sevoflurane on the development of posttraumatic stress disorder (PTSD) in rats and the role of hippocampal glycogen synthase kinase-3beta (GSK-3β).Methods Forty male Sprague-Dawley rats,weighing 250-300 g,aged 2 months,were randomly divided into 4 groups (n =10 each):control group (group C) ; PTSD group (group P) ; sevoflurane group (group S) ; lithium chloride (LiC1) + sevoflurane group (group LS).Establishment of PTSD model:the rats received 15 footshocks (1 mA,1 s shocks with a variable intershock interval of 240-480 s) in room A on the first day.On the second day,the rats received 1 footshock after an adaptation of 192 s in room B where its context was completely different from room A,and then they were removed from room B 32 s later.The rats were placed in room B and observed for 512 s on the third day.Group C did not receive any treatment on the first day in room A,while the other three groups received 15 footshocks in room A.In addition,0.8％ sevoflurane was administrated while training in group S,and group LS received intraperitoneal LiCl 100 mg/kg at 30 min before entering room A,and then the other treatments were similar to those previously described in group S.The ratio of freezing was recorded in room B on the second and third days.Four rats in each group were randomly sacrificed at 2 h after training in room A on the first day and their hippocampal tissues were taken to detect the expression of GSK-3β and phosphorylated GSK-3β (p-GSK-3β).Results Compared with group C,the ratio of freezing was significantly increased on the third day and the p-GSK-3β expression was up-regulated in P and LS groups,and no significant change was found in the parameters mentioned above in group S.Compared with group P,the ratio of freezing was significantly decreased on the third day and the p-GSK-3β expression was down-regulated in group S,and no significant change was found in the parameters mentioned above in group LS.Compared with group S,the ratio of freezing was significantly increased on the third day and the p-GSK-3β expression was up-regulated in group LS.There was no significant difference in the ratio of freezing on the second day and expression of p-GSK-3β between the four groups.Conclusion Sevoflurane can inhibit the development of PTSD in rats,and enhancement of hippocampal GSK-3β activity may be involved in the mechanism.

Objective To investigate the influence of in vitro artificial CO 2 cavity on matrix metallopro-teinase 2(MMP-2), adhesion molecule vascular cell adhesion molecule 1(VCAM-1), and intercellular adhesion molecule 1(ICAM-1)expression in MDA-MB-231 cell.Methods An in vitro artificial CO2 cavity model was es-tablished.MDA-MB-231 cells were exposed to CO2 under the pressure of 7 mmHg for 1, 2 and 4 hours, respective-ly.MMP-2 concentration was measured by enzyme linked immunosorbent assay (ELISA).VCAM-1 and ICAM-1 ex-pression were measured by flow cytometry 0, 24, 48 and 72 hours after CO2-insufflation.Hypoxia group was ex-posed to 0 mmHg helium for 1 h, and the control group was exposed to 37℃incubator only .Results Compared with that in the control group , MMP-2 expression in the 1, 2 and 4 hours treatment group was significantly elevat-ed at 0 hour(F=15.045, P<0.05), and the MMP-2 expression in the 2 hours CO2 treatment group was signifi-cantly elevated after 24 hours compared with that in the control group and 1, and 4 hours CO2 treatment group (F=5.976, P<0.05).The VCAM-1 expression was significantly elevated at 0 hour and after 24 hours in the 1, 2 and 4 hours CO2 treatment group compared with that in the control group ( F1 =18.321, F2 =20.443, P<0.05), and significantly declined after 72 hours in the 4 hours CO2 treatment group compared with that in 1, and 2 hours CO2 treatment group(F=15.045,P<0.05).ICAM-1 expression was significantly elevated at 0 hour in hypoxia group and 1, 2, 4 hours CO2 treatment group compared with that in the control group , Meanwhile it was higher in 2 hours CO2 treatment group than in 1 hours and 4 hours CO2 treatment group(F=73.765, P<0.05). ICAM-1 expression was significantly elevated after 24 hours in 2 and 4 hours CO2 treatment group compared with that in the control group and 1 hour CO2 treatment group(F=46.322, P<0.05), and it was significantly elevat-ed after 48 hours in 2 hours CO2 treatment group compared with that in the control group and 1, and 4 hours CO2 treatment group(F=22.315, P<0.05).Conclusion The expression of MMP-2, adhesion molecule VCAM-1, and ICAM-1 in MDA-MB-231 cells is elevated after exposure to artificial 7 mmHg CO2 cavity, and CO2 cavity of mastoscopy may modulate the metastasis capacity of breast tumor cells .

Objective To explore the clinical efficacy of laparoscope-assisted transanal total mesorectal excision (La-TaTME) for middle-low rectal cancer.Methods The retrospective cross-sectional study was conducted.The clinical data of 16 patients with middle-low rectal cancer who underwent La-TaTME in the Peking University Third Hospital from August 2015 to August 2016 were collected.Sequential surgery of La-TaTME was applied to patients in the same team,with laparoscopic surgery first and then transanal surgery.Observation indicators:(1) operation and postoperative recovery situations:conversion to open surgery,anastomosis method,operation time,volume of intraoperative blood loss,intraoperative complications,time for out-of-bed activity,time for liquid diet intake,postoperative complications and duration of postoperative hospital stay.(2) postoperative pathological situations:length of surgical specimen,tumor diameter,distance from tumor to resected distant intestinal canal,complete degree of mesorectum,circumferential resection margin,pathological T stage,pathological N stage,number of lymph node detected and tumor cell differentiation.(3) follow-up.Patients in stage Ⅲ-ⅣV of TNM stage of RC underwent postoperative adjuvant chemotherapy.Follow-up using outpatient examination was performed once every 3 months postoperatively to detect the patients' survival and tumor recurrence up to December 2016.Measurement data were represented as M (range).Results (1) Operation and postoperative recovery situations:all the 16 patients underwent successful La-TaTME without conversion to open surgery,including 10 with colorectal anastomosis,3 with colon-canalis analis anastomosis and 3 with permanent colostomy.Operation time and volume of intraoperative blood loss were 290 minutes (range,215-420 minutes) and 50 mL (range,30-100 mL),respectively.One patient had intraoperative complication,showing broken ends ischemia of sigmoid colon after dragging out resected rectum from the anus,following free splenic flexure of colon,about 5 cm ischemic sigmoid colon were resected,and descending colon-rectum anastomosis was performed.Time for out-of-bed activity and time for liquid diet intake were 1 days (range,1-3 days) and 2 days (range,1-9 days),respectively.Among 3 patients with postoperative complications (Ⅱ stage of ClavienDindo),2 with incomplete intestinal obstruction were improved by gastrointestinal decompression and total parenteral nutrition,and 1 with presacral infection was improved by drainage and antibiotic therapy.Duration of postoperative hospital stay was 7 days (range,5-21 days).(2) Postoperative pathological situations:length of surgecal specimen,tumor diameter and distance from tumor to resected distant intestinal canal were respectively 18.0 cm (range,12.0-24.0 cm),3.5 cm (range,0.5-6.8 cm) and 2.5 cm (range,1.0-5.0 cm).Evaluation of mesorectum of surgical specimen:14 patients had complete mesorectum of surgical specimen and 2 had nearly complete mesorectum.There was no residual tumor at circumferential resection margin,proximal and distal ends.Pathological T stage of 16 patients:T0 (pathological complete response after neoadjuvant therapy),T1,T2 and T3 stages were found in 1,1,4 and 10 patients,respectively.Pathological N stage:12,2 and 2 patients were detected in N0,N1 and N2 stages,respectively.Number of lymph node detected was 16 (range,6-32).Tumor cell differentiation:no tumor cell (pathological complete response after neoadjuvant therapy),high-,moderateand low-differentiated tumors were respectively detected in 1,2,7 and 6 patients.(3) Follow-up.All the patients were followed up for 12 months (range,4-16 months).There were no local tumor recurrence or distant metastasis and death.Conclusion La-TaTME may be a new,safe and effective resection for middle-low rectal cancer.