Collagen (Coll), as the basic material of matrix scaffolds for cell growth, has been widely used in the field of tissue engineering and regenerative medicine. In this study, collagen protein was modified by L-lysine (Lys), and cross-linked by genipin (GN) to prepare the L-lysine-modified collagen (Lys-Coll-GN) scaffolds. Microstructure, pore size, porosity, stability and biocompatibility of Lys-Coll-GN scaffolds were observed. The results showed that the bond between L-lysine and collagen protein molecule was formed by generating amide linkage, and mouse embryo fibroblasts proliferation was not inhibited in the Lys-Coll-GN scaffolds. In the multiple comparisons of Coll-scaf- folds, Coll-GN scaffolds and Lys-Coll-GN scaffolds, Coll-scaffolds was the worst in mechanical characteristics while the highest in biodegradation rate. Compared to Coll-GN scaffolds, Lys-Coll-GN scaffolds had more fiber structure, higher interval porosity (P<0. 01). Although the tensile stress of Lys-Coll-GN scaffolds reduced significantly, its e- longation length extended when the scaffolds was fractured (P<0. 01). The percentage of Lys-Coll-GN scaffolds residual weight was lower than that of Coll-GN scaffolds after all the scaffolds were treated by collagenase for 5 days (P<0. 01). This study suggested that Lys-Coll-GN scaffold had good biocompatibility, and it improved the mechanical property and degradation velocity for collagen-based scaffold. This study gave a new predominant type of tissue engineering scaffold for the regenerative medicine.
Animals ; Biocompatible Materials ; chemistry ; Cell Proliferation ; Collagen ; chemistry ; Cross-Linking Reagents ; Fibroblasts ; cytology ; Iridoids ; chemistry ; Lysine ; chemistry ; Mice ; Porosity ; Tissue Engineering ; Tissue Scaffolds

Objective:To investigate the prognosis of patients with liver metastasis or local recurrence after radical resection of rectal cancer,in order to provide reference for the further screening of high-risk patients for the precise therapeutic methods.Methods:The clinicopathological factor and follow-up data of 485 patients who underwent surgical treatment for rectal cancer from March 2005 to January 2016 were retrospectively analyzed.Including 75 liver metastasis and 32 local recurrence.The prognosis were compared between the patients with liver metastasis and with local recurrence.Results:The difference was statistically significant in CEA level,primary tumor position,surgical methods,tumor cell differentiation,tumor infiltration depth between liver metastasis and local recurrence after radical resection of rectal cancer (P＜0.05).The 1-,3-and 5-year overall survival rates were 76.6％,53.1％ and 18.8％ respectively of patients with liver metastasis,The 1-,3-and 5-year overall survival rates were 81.3％,62.5％ and 37.5％ respectively of patients with local recurrence,the difference was statistically significant(P＜0.05).Conclusion:There were different clinicopathological characteristics of patients between liver metastasis and local recurrence.The prognosis of patients with local recurrence was better than patients with liver metastasis.

Objective To research the effects of progressive time phase selective light (PTPSL)therapy on shift work disorder patients.Methods 24 shift work disorder patients were randomlv divided into PTPSL group and control group.Both groups received treatment for 12 days including 6 days during the night shift time and 6 days during the day shift time continuously.The clinical effects were evaluated by Karolinska Sleepiness Scale (KSS),Leeds sleep evaluation questionnaire (LSEQ) and core body temperature rhythm.Results (1) During the night shift time,PTPSL group patients were better in score LSEQ and KSS(discerned for 39.80 ± 2.89,6.59 ±0.48) than control group' s(discerned for 36.86 ± 3.88,6.98 ± 2.03) ; temperature curve peak phase in PTPSL group moved backward greater than that of control group((8.35 ± 0.82) h vs (7.45 ± 1.05) h) ; and the difference was statistical significance(P ＜ 0.05).During the day shift time,PTPSL group patients were better in score LSEQ and KSS(discerned for 45.57 ± 2.77,6.33 ± 0.48) than control group' s (discerned for 40.35 ± 4.19,6.82 ± 1.03),temperature curve peak phase in PTPSL group moved forward greater than that of control group((7.37 ± 0.94)h vs (6.31 ±1.37)h),and the difference was statistical significance(P＜0.05).Conclusion PTPSL can adjust shift sleep disorder patients' mussily sleep rhythm system in order,and make the inner rhythm svstem and external schedules tend to be consistent,and provides a new treatment choice for shift sleep disorders.

Objective To investigate the demethylation and changes in gene expression of programmed death receptor-1 ( PD-1) caused by methylation inhibitor 5- azacytidine (5-Zac) in lymphocyte series Molt-4 cells and its mechanism. Methods Molt-4 cells were cultured in different concentrations of 5-Zac(0, 5, 10 Umol/L)for 72 h, ratio of cell expressing PD-1 and apoptosis rate were detected by FCM, transcription of PD-1 gene mRNA was detected by RT-PCR. Molt-4 cell DNA of all groups were disposed by sodium bisulfite, PD-1 gene promoter fragment binded with transcription factor Brn-2 was amplified by PCR,these amplification fragments were transformed into E. coli. Positive clones were selected by sequencing,methylation status of the fragments binded with transcription factor Brn-2 was examined. Results S-Zac could increase the PD-1 expression of Molt-4 cells. PD-1 expression rate in 0 μmol/L 5-Zac( 1. 13%±0.01% ) treated cells was found more lower than that in both 5 μmol/L and 10 μmol/L 5-Zac treated cells (18. 96% ±1. 87% , 63. 09% ± 6. 25% , P < 0. 05 ) , and they showed concentration-dependent (P <0.01). Cells apoptosis rate and PD-1 mRNA expression were also observed increased significantly with 5-Zac treating. Demethylation probability of CG points showed significant difference between transcription factor Brn-2 binding site and other four locations (P < 0.05 ). Conclusion 5 -Zac inhibits cell grouth in human lymphoid cell series Molt-4 by inducing PD-1 gene expression and promoter demethylation. PD-1 gene promoter binding transcription factor Brn-2 fragment CG point demethylation may be one of the important mechanisms in 5-Zac treated Molt-4 cells.

Bacterial Typing Techniques ; methods ; Brucella ; classification

Objective:To investigate the purified methods of human anti-D antibody from IgG contained anti-D. Methods:The IgG was separated by the column ion-exchange chromatography(CIEC) from the plasma in which the content of anti-D was 0.814 ?g/ml. Then the IgG preparation contained anti-D was purified by the affinity chromatography(AC) with the O group, RhD positive red blood cell (genotype CCDee). Results:The content of non anti-D IgG were reduced about 90% by the method of AC and the proportion of anti-D could be significantly increased in the final preperation. The quality of final preparation attained reqirements of national standard of biologics. Conclusion:This method is able to purify anti-D from IgG contained anti-D and offer a reference for plasma products.

Objective To study the clinical efficacy of percutaneous transluminal angioplasty (PTA) and vacuum sealing drainage technique on patients with diabetic foot and arteriosclerosis obliteration of the lower extremity.Methods Sixty-four patients were divided into two groups.Group A (32 patients) were treated by traditional methods,while 32 patients in Group B treated by vacuum sealing drainage technique in addition to percutaneous transluminal angioplasty.Results One week after surgery,ABI in group A was 0.53 ±0.17 vs 0.85 ±0.27 in group B,P =0.00.Ulcer healing rate increased from 31.25％ in group A to 81.25％ in group B,P ＜0.01.The amputation rate decreased from 56.25％ in group A to 18.75％ in group B,P =0.00.Claudication distance extended and ulcer recurrence rate in group A was 40％,while in group B it was 0,P =0.00.Conclusion The combination of PTA and vacuum sealing drainage significantly improves the prognosis of patients with diabetic foot and arteriosclerosis obliteration of the lower extremity.

AIM: To investigate the effect of phospholamban antisense RNA (asPLB) on the activity of sarco-endoplasmic reticulum (SR) Ca 2+-ATPase, and the change of intracellular free Ca 2+ concentration ([Ca 2+]i) in rat cardiomyocytes by adeno-associated virus(AAV) vector. METHODS: rAAV-asPLB and rAAV-LacZ were constructed by AAV Helper-Free System. RT-PCR and Western blotting were used to determine the mRNA and protein expression of PLB. The activity of SR Ca 2+-ATPase and the [Ca 2+]i were measured. RESULTS: Compared to controls, the PLB mRNA and protein expression reduced in rat cardiomyocytes transfected with rAAV-asPLB. The activity of Ca 2+-ATPase was increased. In rest state, the level of [Ca 2+]i in rAAV-asPLB transfected group was decreased. The level of [Ca 2+]i was increased when induced by isoproterenol. CONCLUSION: rAAV-asPLB vector disrupts the expression of PLB, enhances the activity of Ca 2+-ATPase, reduces the resting [Ca 2+]i and enhances the isoproterenol-induced [Ca 2+]i.

Objective: To investigate the clinicopathological features, diagnosis and differential diagnosis of gastrointestinal glomus tumors (GIGT). Methods: Totally 15 cases of GIGT were collected at the First Affiliated Hospital, Zhengzhou University, from January 2011 to June 2018. The clinicopathological features, immunophenotype, BRAF V600E mutation and prognosis were retrospectively analyzed. Results: The 15 patients′ age ranged from 37 to 59 years(median 49 years, mean 50 years). Eleven patients presented with intermittent abdominal pain and distention, three showed antral space-occupying lesions at physical examination, and one had abdominal pain accompanied by fecal blood. Fourteen tumors were located in the stomach, and one was in the ileum. Imaging showed the gastric glomus tumors were located in the submucosal layer with obvious enhancement in the arterial phase, and the ileum glomus tumor involved the whole layer of intestinal wall causing luminal obstruction. The maximum diameters of the tumors ranged from 1.5 to 3.0 cm (mean 2.3 cm). Grossly, the gastric glomus tumors were solid. Microscopically, the gastric glomus tumors were mostly located in the muscularispropria layer and were vascular. The tumor boundary was distinct but without capsule formation. The tumor cells were round or oval, and showed perivascular hemangiopericytoma-like or solid nest-like structures. The tumor cells were mildly pleomorphic, with rare mitosis and no necrosis. Two tumors had focal calcification, two showed mucosal invasion, two showed vascular invasion and five showed perineural invasion. The ileum glomus tumor was cellular, with prominent cellular atypia, and the mitotic count in hot spots was about 5-6/HPF. Immunohistochemistry showed that SMA and collage Ⅳ were strongly expressed in all the tumor cells; caldesmon and calponin were moderately expressed in some regions, and syn was weakly expressed in 12 cases. The Ki-67 proliferation index in the gastric glomus tumors ranged from 1% to 30% (mean 6%); and that in the ileum glomus tumor was about 70%. BRAF V600E mutations were not detected in any of 15 GIGTs. All patients did not receive radiotherapy or chemotherapy post operatively. Thirteen patients were followed up by telephone for 18-90 months (mean 42 months). Twelve patients with gastric glomus tumors survived without recurrence and metastasis, and the patient with ileum glomus tumor had liver metastasis 15 months after operation. Conclusions Glomus tumors is a rare mesenchymal tumor of the gastrointestinal tract. It should be differentiated from gastrointestinal stromal tumors, neuroendocrine tumor, leiomyoma, solitary fibrous tumor and paraganglioma. Most GIGTs are benign and have good prognosis. More experience is needed to understand the biologic behavior and prognostication of GIGTs.

Objective To investigate the effect of BIS-guided closed-loop target-controlled infusion on perioperative Th1/Th2 balance in elderly patients undergoing laparoscopic radical gastrectomy under different anesthesia and sedation depths. Methods We applied random number table method to divide 73 elderly patients undergoing elective laparoscopic radical gastrectomy into BIS closed-loop target-controlled infusion group with BIS value of 55(group H, n=36) and BIS value of 45(group L, n=37).Intravenous blood samples were collected immediately before surgery (T1), 2h after surgery (T2), 24h after surgery (T3) and 72h after surgery (T4).IL-2, IL-4, IL-6, IL-10, TNF-α and IFN-γ were determined by flow cytometry microsphere array.We compared operation duration, postoperative PACU stay time and postoperative hospitalization time between two groups. Results Compared with T1, IL-6 and IL-10 concentration in group H at T2, T3 and T4 significantly increased (P < 0.05), IL-4 and TNF-α in group H at T4 were significantly increased (P < 0.05), IL-2 and IFN-γ in group H at T2 and T4 were significantly increased (P < 0.05), the concentration of IL-6 and IL-10 in group L at T2, T3 and T4 were increased (P < 0.05), IL-2 in group L at T4 was decreased (P < 0.05), and IFN-γ/IL-6 in two groups were decreased at T2, T3 and T4(P < 0.05).Compared with group L, IL-6 and IL-10 in group H were significantly increased at T2(P < 0.05), IFN-γ/IL-6 in group H was significantly decreased (P < 0.05), IL-2 and IL-10 in group H were significantly increased at T4(P < 0.05).Operation duration, postoperative PACU stay time and postoperative hospitalization time had no statistical significance between two groups (P > 0.05). Conclusion The anesthesia and sedation depth of BIS-guided closed-loop target-controlled infusion set at 45 is better than 55 in maintaining Th1/Th2 balance in elderly patients undergoing laparoscopic radical gastrectomy, but it has no obvious effect on long-term prognosis.