Objective: To investigate the anti-tumor effect of lymph oc yte-mediated immunity induced by Tca8113 cell lysate-pulsed dendritic cells(DC s). Methods: Human peripheral blood monocytes were cultured in R PMI 1640 medium countaning ?=10% FBS with GM-CSF + IL-4 + either TNF-? or n ot. Control cells were cultured without any cytokine. Nonadherent cells were har vested, and continuously incubated with Tca8113 cell lysate made by freez-thaw method, and then the cells were cultured together with autologous lymphocytes. T he activated lymphocytes were co-cultured with Tca8113 cells at E∶T ratio of 1 00∶1. The cells were observed morphologically and assessed by immunocytochemis try before and after the pulse, while the survival rate of Tca8113 cells was stu died by MTT method. All results were analyzed statistically. Results: (1) Typical DCs were generated with higher expression of CD83 in the group s with TNF-? treatment, or with tumor antigen stimulation. (2) The effective l ymphocytes induced by DCs inhibited the proliferation of Tca8113 cells. Conclusion: DCs generated from monocytes in the presence of GM-CSF an d IL-4 can develop to be mature after tumor antigen pulse. DCs induce the lymph ocyte-mediated suppression effect on Tca8113 cells; TNF-? can accelerate DC m ature.

ObjectiveTo evaluate the curative effect of reconstruction of the hand and foot defects with bones and tendons exposure using free medial sural artery perforator flap(MSAP). MethodsRadiographs of 2 cadavers injected with a modified lead oxide-gelatin mixture were digitally analyzed. Between April 2007 and December 2010, thirty-four patients with soft tissue defects in the distal limb were treated with the free MSAP flap transplantation. The sizes of the defect ranged 6 cm × 4 cm-13 cm × 8 cm, and the flaps ranged 7 cm× 5 cm-14 cm × 9 cm. These clinical cases included 25 hands and 9 feet, all of them with bones and tendons exposure.In these defects,twenty-two were clean,twelve got infections.In our cases, twenty-three flaps were nourished with single perforator vessel and else 11 with two;perforator vessel fifteen flaps were dissected one superficial vein to anastomose with that of the recipient sites in addition to accompanying vein anastomosis;The sensation of 9 flaps recovered the hands were reconstructed with cutaneous nerve anastomosis. ResultsA partition of the calf skin blood vessels,and three-dimensional reconstruction image of the sural artery were obtained.All flaps survived,five of them appeared partially violet and bubbles. Followed up 6-21 months, the cosmetic results were satisfactory and without apparent bulkiness.The flap colors were similar to recipient sites. The flap senses reconstructed with neural anastomosis recover to S2-S3. ConclusionThe new flap is very suitable to repair the soft tissue defect in the distal limbs,because the fairly constant perforator vessel,the reliable blood supply and the cosmetic shape of the MSAP flap are all advantages of it in addition to no damage to low leg chief artery and gastrocnemius.

Objective To provide anatomical landmarks with which to facilitate flap dissection,we studied the perforator artery of the dorsal forearm including its source,quantity,origination,caliber,variation and pedicle length. Methods Ten fresh cadavers were injected with a modified lead oxide-gelatin mixture,and three-dimensional graphics of the perforator vessels of the dorsal forearm were reconstructed with a computed tomography. In addition, twenty upper extremity specimens were injected with red latex via the axillary artery.The integument of the forearm was dissected,and perforators were identified,including type,course,size and location were documented.Surface areas were measured with Scion Image. Results The average number of the posterior interosseous artery cutaneous perforators in the dorsal forearm was (5±2),the average outer diameter of the perforator artories was (0.5 ± 0.1) mm,and the pedicle length was (2.5 ±0.2) cm.The average cutaneous vascular territory was (22.0 ± 15.0) cm2.The dorsal branch of the anterior interosseous artery dispersed on the wrist dorsum or the distal third of the dorsal forearm. It's average diameter was 0.8 mum. Conclusion The free transplantation of the posterior interosseous perforator artery flaps or rotary flap pedicled by the dorsal branch of the anterior interosseous artery for defect reconstruction are feasible.

Objective To evaluate the clinical outcome of the method of repairing donor site of foot after improved toe-to-thumb reconstruction utilizing superficial circumflex iliac artery perforator (SCIAP) chimeric flap.Methods Fourteen cases of thumb defect were recruited from April,2012 to January,2016.According to Gu Yudong's classification,5 cases met the criterion of type Ⅰ,4 cases met the criterion of type Ⅱ,and 5 cases met the criterion of type Ⅲ.For type Ⅰ,the thumb was reconstructed with the great toe wrap-around flap.For type Ⅱ and Ⅲ,the thumb was reconstructed by the combined tissue with mutual artery (great toe wrap-around flap,and the bonetendon tissue of the second toe).All the donor sites of foot were repaired utilizing SCIAP chimeric flap.Results All the reconstructed thumbs survived.Among 14 free flaps of donor site,1 case suffered venous crisis and survived after exploration and rescue surgery.Dorsal skin necrosis of the second toe was found in 1 case,which was healed by local skin flap transposition.All patients were followed-up ranged from 3 to 30 months (averaged at 16 months).In spite of slightly bloated,the color and texture of all the flaps' was satisfied,and the average healing time of the bone in the donor sites was 2.5 months.All patients did not feel painful and had no adverse effect when walking and running.Three months after the operation,5 slightly bloated flaps in the donor sites under went flap plastic and achieved better appearance.On the part of iliaca,there was only one inconspicuous linear scar without any discomfort.Conclusion Repairing donor site of foot after improved toe-to-thumb reconstruction utilizing SCIAP chimeric flap was an ideal method.Using this method,the reconstructed thumb can achieve good appearance and function,all the toes of donor site were reserved,and the disability of the donor site is minimized.

In this study the blood sample was collected from eighty-six athletes in Tibetan mountaining team and Tibetan mountaining sports school and ninety healthy Han nationality people in Guangdong province, and genomic DNA was extracted from peripheral leukocyte. The allele frequency distribution and the genotypes combination distribution of hypoxia-inducible factor-la gene (HIF-1alpha)exonl2 C1772T and G1790A were examined by restriction fragment length polymorphism PCR (PCR-RFLP) in order to evaluate the association of single nucleotide polymorphisms (SNPs) of HIF-1alpha C1772T and G1790A with hypoxic acclimation in high altitude in Tibetans. The results indicated that the genotype frequency of HIF-1alpha C1772T in Tibetan and in Han nationality was 13.95% versus 16.67% in genotype CC, 38.37% versus 41.11% in genotype CT and 47.68% versus 42.22% in genotype TT. No significant difference in CC, CT and TT genotype frequency of HIF-1alphaC1772T was shown between Tibetans and Han nationality respectively, while GA genotype frequency of HIF-la G1790A in Tibetans was higher than that in Han nationality. The GA genotype of HIF-1alpha G1790A may be involved in the hypoxic acclimation of high altitude , and it is worth of deep-going investigation.
Adaptation, Physiological ; genetics ; physiology ; Adolescent ; Adult ; Aged ; Altitude ; Base Sequence ; Female ; Humans ; Hypoxia ; genetics ; Hypoxia-Inducible Factor 1, alpha Subunit ; genetics ; Linkage Disequilibrium ; Male ; Middle Aged ; Molecular Sequence Data ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length ; Polymorphism, Single Nucleotide ; genetics ; Tibet ; Young Adult

This investigation was conducted to explore the relationship between the polymorphism of gene of glucose transport 1(GLUT1) and the human body adaptation to high altitude hypoxia environment. The data on glucose transport 1 gene polymorphism in Tibetan mountaineers (high altitude group) were analyzed and compared with the data from the level-land Guangdong Hans (control group). The genotype of 86 Tibetan mountaineers and 90 level-land Hans as controls were tested with polymerase chain reaction followed by restriction fragment length polymorphism analysis for GLUT1 gene. The results showed that, in the high altitude mountaineer group, the frequencies of +22999 locus genotypes GG, GT and TT were 44.2%, 46.5% and 9.3% respectively, and such frequencies in the control group were 66.7%, 31.1% and 2.2% respectively. The frequencies of + 22999 polymorphic genotypes and alleles showed statistically significant difference between the high altitude group and the control group (P<0.05). Genetic single nucleotide polymorphism in GLUT1 G+22999T may be associated with the adaptation to high altitude hypoxia.
Adaptation, Physiological ; genetics ; Adolescent ; Adult ; Alleles ; Altitude ; Base Sequence ; Female ; Genotype ; Glucose Transporter Type 1 ; genetics ; Humans ; Hypoxia ; genetics ; Male ; Middle Aged ; Molecular Sequence Data ; Polymerase Chain Reaction ; methods ; Polymorphism, Restriction Fragment Length ; Tibet ; Young Adult

Background and Objectives: Sheep-induced pluripotent stem cells (siPSCs) have low reprogramming efficiency, thereby hampering their use in biotechnology and agriculture. Several studies have shown that some microRNAs play an important role in promoting somatic reprogramming in mouse and human. In this study, we investigated the effect of miR-200c-141 on somatic reprogramming in sheep and explored the mechanism of promoting the reprogramming. Methods: and Results: The lentivirus system driven by tetracycline (TET)-on carrying Oct4, Sox2, c-Myc, Klf4, Nanog, Lin28, hTERT, and SV40LT (OSKMNLST) could reprogram sheep kidney cells into pluripotent cells. Overexpression of miR-200c-141 in combination with OSKMNLST could significantly improve the efficiency of sheep iPSC generation (p＜0.01). Sheep iPSCs derived from miR-200c-141 showed embryonic stem cell (ESC)-like pluripotent properties, were positive for alkaline phosphatase and some pluripotent markers by quantitative real-time PCR (qRT-PCR) and immunofluorescence, and were able to differentiate into three germ layers in vitro. Oar-miR-200c was transfected into HEK293FT cells and was able to target the zinc finger E-box-binding homeobox 1 (ZEB1) 3’UTR using dual luciferase reporting analysis. Overexpression of oar-miR-200c in SKCs significantly reduced the expression of ZEB1, but increased the expression of E-cadherin by qRT-PCR and western blotting analysis. Conclusions These results suggest that miR-200c-141 can promote the reprogramming of sheep somatic cells to iPSCs, and oar-miR-200c targeted ZEB1 3’UTR, significantly decreased expression of ZEB1, and increased expression of E-cadherin. Oar-miR-200c may improve the MET process by affecting the TGF-β signaling pathway, thus improving the efficiency of somatic cell reprogramming in sheep.

OBJECTIVETo mimic oral squamous cell carcinoma (OSCC) cell hypoxia by using chemical agent CoCl2 and to investigate its biological behaviour.

METHODSOral squamous cell carcinoma cell lines HSC-3 and SCC-4 were exposed to different concentration of CoCl2. HSC-3 and SCC-4 cells were treated with 50, 100, 150, 200 µmol/L CoCl2. Expression of hypoxia inducible factor-1α (HIF-1α), vascular endothelial growth factor (VEGF) and B-cell lymphoma-2 (BCL-2) were measured by real time polymerase chain reaction (PCR) and Western blotting in both mRNA and protein level. Cell proliferation, cell apoptosis and cell cycle were detected to analyze its biological behaviour. Both wound healing and Transwell assay were applied to test the ability of cell igration.

RESULTSThe result showed that after treatment of 150 µmol/L CoCl2 for 24 h, mRNA level of HIF-1α, VEGF and Bcl-2 was increased by 6.00 ± 0.20, 5.40 ± 0.40, 5.40 ± 0.30 (SCC-4); 5.60 ± 0.30, 5.20 ± 0.60, 5.80 ± 0.40(HSC-3). OSCC cells treated with 150 µmol/L CoCl2 for 24 h were collected. Compared with control group, the growth rate of cells was significantly decreased, P value was less than 0.05 (when HSC-3, SCC-4 cultured for 2 and 3 days). The apoptosis of OSCC cells was increased when treated with 150 µmol/L CoCl2 for 24 h:HSC-3 2.25% (control group) and 5.82% (treatment group); SCC-4 2.58% (control group) and 10.27% (treatment group). The migration ablility of OSCC cells was decreased when using 150 µmol/L CoCl2 for 24 h. The migration area ratio was (31.5 ± 2.3) % (HSC-3), (29.1 ± 1.5) % (SCC-4) in control group and (18.3 ± 1.9) % (HSC-3), (13.2 ± 0.8)% (SCC-4) in treatment group (P < 0.05).

CONCLUSIONSThe hypoxic cell model of OSCC could be induced by CoCl2. The expression level of hypoxic markers was up regulated significantly and the cells biological behaviour changed including decreased cell proliferation, increased apoptosis and decreased migration.

Apoptosis ; Blotting, Western ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Cell Cycle ; Cell Hypoxia ; Cell Line, Tumor ; Cell Movement ; drug effects ; Cell Proliferation ; Cobalt ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; metabolism ; Mouth Neoplasms ; metabolism ; pathology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; RNA, Messenger ; metabolism ; Up-Regulation ; Vascular Endothelial Growth Factor A ; metabolism

Chylous fistula after abdominal operation is a rare complication which easily lead to malnutrition or immunosuppression and even cause death due to constant loss protein and lymphocytes.The therapeutics of the chylous fistula after abdominal operation includes conservative treatment,surgery and special management modalities at present.The most of the patient can be cured by non-surgical treatment,which should be considered as the first treatment regimen,surgery should be choosen if it failed.Only the obove methads have failed can we take the special treatments cound be taken into consideration.In this paper,new progress in diagnosis and therapeutics of the chylous fistula after abdominal operation will be reviewed.

Objective:To investigate the computed tomography (CT) features of gastric neuroendocrine neoplasm (GNEN).Methods:The retrospective and descriptive method was conducted. The clinicopathological data of 30 GNEN patients who were admitted to two domestic medical centers (13 cases in Wenzhou Hospital of Traditional Chinese Medicine and 17 cases in Wenzhou People′s Hospital) from January 2010 to December 2018 were collected. There were 23 males and 7 females, aged (62±4)years, with a range from 27 to 78 years. The patients underwent abdominal CT plain scan and dynamic enhancement scan. Two associate chief radiologists with more than 20 years of work experience observed and analyzed all the images respectively. Observation indicators: (1) CT examination; (2) treatment and postoperative pathological examination; (3) follow-up. Follow-up was conducted using outpatient examination and telephone interview to detect the survival of patients up to December 2018. Measurement data with normal distribution were represented as Mean± SD. Measurement data with skewed distribution were described as M (range). Results:(1) CT examination: of the 30 patients, 14 had the tumor located in the fundus of stomach, 10 had the the tumor located in the body of stomach, and 6 had the tumor located in the antrum. The tumor was elliptical in 18 cases and irregular in 12 cases. There were 15 cases of endogenous type, 13 cases of exogenous type, and 2 cases of intramural type. Patients with G1 neuroendocrine tumor had the maximum diameter of (6.8±1.6)cm, of which 4 cases had the maximum diameter less than 5.0 cm and 4 cases had the maximum diameter of 5.0 to 10.0 cm. Patients with G2 neuroendocrine tumor had the maximum diameter of (8.3±2.7)cm, of which 1 case had the maximum diameter less than 5.0 cm, 4 cases had the maximum diameter of 5.0 to 10.0 cm, and 2 cases had the maximum diameter greater than 10.0 cm. Patients with G3 neuroendocrine carcinoma had the maximum diameter of (17.8±2.2)cm, of which 6 cases had the maximum diameter of 5.0 to 10.0 cm and 9 cases had the maximum diameter more than 10.0 cm. The tumor showed swelling growth in 14 cases and invasive growth in 16 cases. The tumor boundary was clear in 14 cases and unclear in 16 cases. CT plain scan showed homogeneous tumor density in 10 cases and heterogeneous density in 20 cases. Nine patients had iso-density in the tumor parenchymal part, and the CT value was (34.0±3.5)HU. In the 18 cases of low density, the CT value was (16.6±1.4)HU. In the 3 cases of high density, the CT value was (45.3±3.6)HU. Of the 30 patients, 21 cases had small punctate or small round necrotic cyst lesions in the tumor, 10 cases had mesenteric lymph node, peritoneum, liver metastasis and adjacent omentum invasion; 17 cases had abdominal effusion. In the CT enhancement examination, 12 cases showed mild enhancement, and the CT value was (56.5±6.3)HU; 15 cases showed moderate enhancement, and the CT value was (66.0±5.4)HU; 3 cases showed significant enhancement, and the CT value was (76.6±5.8)HU. Seven cases showed homogeneous enhancement and 23 cases had heterogeneous enhancement. There were 8 cases with tortuous vessels. (2) Treatment and postoperative pathological examination: of the 30 patients, 10 cases with mesenteric lymph nodes, peritoneum, liver metastasis and adjacent omentum invasion underwent radical total gastrectomy; 14 cases without surrounding tissue invasion or metastasis underwent radical subtotal gastrectomy; 6 cases with tumor diameter less than 4.0 cm and without surrounding tissue invasion or metastasis underwent endoscopic resection. All the 30 patients were confirmed GNEN by postoperative pathological examination, including 8 cases of G1 neuroendocrine tumor, 7 cases of G2 neuroendocrine tumor, and 15 cases of G3 neuroendocrine carcinoma. Results of immunohistochemical staining showed that 30 patients were positive for synaptophysin, 23 were positive for chromogranin A, and 9 were positive for cytokeratin. (3) Follow-up: all the 30 patients were followed up for 10-80 months, with a median follow-up time of 39 months. The 5-year survival rate of 30 patients was 43.3% (13/30). The 5-year survival rates were 6/8, 3/7 and 4/15 of patients with G1 neuroendocrine tumor, G2 neuroendocrine carcinoma, and G3 neuroendocrine carcinoma.Conclusions:GNEN has the main manifestation as abdominal pain, with G3 as pathological classification, which is common in fundus and body of stomach. The CT findings of GNEN are characterized by swelling or infiltrating growth and round or irregular low-density masses. Tumors are prone to cystic transformation, and showed the mildly to moderately heterogeneous enhancement.