Objective To investigate the cellular immune response to the recombinant DNA vaccine CRT120/HPV6bE7 in mice. Methods The recombinant encoding HPV6bE7, linked with CRT120, was constructed in pcDNA3.1 eukaryotic vector with the report gene GFP. The pcDNA3.1-GFP -HPV6bE7 was transfected into B16 cells by a lipofectamine kit. The C57BL/6 mice were vaccinated with recombinant DNA plamids. The T-cell phenotype in the peripheral blood lymphocytes of the immunized mice was measured by flow cytometry. The CTL activity of the lymphocytes from the spleens and lymph nodes, and the levels of IL-2 and IFN-? were analyzed. Results The constructed recombinant plasmid CRT120/HPV6bE7 was analyzed. Positive transfected cell clones were established and could stably express the target gene HPV6bE7. Compared with HPV6bE7, CRT120/HPV6bE7 plasmid enhanced to a greater extent CD8+ T-lymphocyte differentiation, the number of TCR?? T-lymphocytes and the levels of IL-2 and IFN-? (all P

OBJECTIVE: To evaluate the advantage of Gukangling Decoction (GKLD), a compound traditional Chinese herbal medicine, combined with technetium [(99)Tc] methylene diphosphonate injection ((99)Tc-MDP) in treating osteoporosis in rabbits. METHODS: A rabbit model of osteoporosis was established by intramuscular injection of dexamethasone (DX). Fifty-six rabbits were divided into 8 groups: Group A, B, C, D, E, F, G and H. Rabbits in groups A and B were intramuscularly injected normal saline as normal control, groups C and D were untreated groups, rabbits in group E were treated by (99)-MDP, rabbits in group F were treated by aminodiphosphate, rabbits in group G were treated by GKLD, and rabbits in group H were treated by (99)-MDP and GKLD. Rabbits in groups A and C were executed to demonstrate the establishment of the rabbit model of osteoporosis at the 8th week of experiment. Rabbits in the other six groups were executed after 16-week experiment (8-week treatment), and then bone structure and cell shape were observed by electron microscope, X-ray, CT and emission computed tomography (ECT). Bone density, biomechanical parameters, the levels of bone specific alkaline phosphatase (BALP) and bone Gla protein (BGP) were measured too. RESULTS: After 8-week of intramuscular injection of DX, the bone trabecula in group A were regular and showed normal configuration, while the bone trabecula in group C were sparse, ruptured and showed damaged form. The bone density and biomechanical parameters in group A were higher than those in group C, indicating that the rabbit model of osteoporosis was established successfully. At the 9th week of experiment, the results of cell pathology in group D showed that the bone trabeculas were sparse, ruptured, defected or had hollow section, but the bone trabeculas in group B were regular and dense. The bone trabeculas in groups H and E were restored, and were thicker than those in group D. The bone quality in groups H and E was better than group D significantly, the bone quality in group F was better than group G, and the bone quality in group G was better than group D slightly. CONCLUSION: GKLD combined with (99)-MDP had superiority in treating osteoporosis of rabbits as compared with the respective single therapy.

Objective To summarize clinical experience of one-station therapy for infected seriouslyischemic diabetic foot.Methods The clinical data of 15 patients (15 diseased limbs in total) with infected seriously-ischemic diabetic foot,who were admitted to authors' hospital during the period from June 2015 to April 2016 to receive treatment,were retrospectively analyzed.For all patients,one-station sequential therapy was carried out,which included endovascular revascularization (EVR) to open occluded vessel,surgical debridement and closed negative pressure wound drainage and antiseptic moisturizing wound dressing.The healing rate of infected wound and the limb salvage rate were evaluated.Results The 15 patients included 10 males and 5 females,with a median age of 77 years old.Lower extremity angiography showed that multiple segmental lesions of lower limb were detected in 13 patients and simple leg lesions in 2 patients.According to TASC Ⅱ update classification,leg artery disease of grade D was observed in 13 patients and artery disease of grade C in 2 patients.After EVR therapy,at least one branch of leg arteries was reopened in 14 limbs.Intact arterial arch of pedal-plantar loop (PPL) was seen in 6 patients,semi-arterial arch in 7 patients,and absent of arterial arch in 2 patients.After surgical debridement,the wound was washed by using negative pressure wound therapy (NPWT) device as well as serf-made washing equipment.The time to control wound infection was (7.85±2.84) days.After discharge,the patients were followed up every 3-4 days,at the same time wound dressing exchange with antibacterial moisturizing sulfadiazine silver lipid hydrogel was conducted.Wound healing was achieved in 12 patients,and the mean healing time was (3.70±0.87) months.The wound failed to heal in 3 patients,among them below knee amputation had to be performed in 2 patients (13.3％,both patients showed absent of arterial arch of PPL),and the remaining one patient died of cardiovascular event.Statistically significant difference in PPL pathological changes existed between wound healing group and wound un-healing group (P=0.006 7).Conclusion The treatment of infected seriouslyischemic diabetic foot is rather complicated.Being one-station therapy,the sequential managements,which include EVR,NPWT device together with washing equipment and use of antibacterial moisturizing wound dressing,can effectively increase the blood supply to the affected limb,shorten the time to control infection and lower amputation rate.Therefore,one-station therapy should be regarded as the preferred method for infected seriously-ischemic diabetic foot.

Objective To explore the value of wound blush in predicting patients' ulcer healing whom with critical limb ischemia after revascularization.Methods Retrospectively analyze the clinical data of 173 cases of critical limb ischemia with ischemic ulcers under thetreatment of endovascular therapy followed the concept of angiosome.According to the condition of wound blush after endovascular therapy,by compared the difference of limb salvage rate and ulcer healing time,and try to analyze the value of wound blush in predicting ulcer healing in patients.Results Included in the study with a total of 173 cases (173 limbs),group wound blush(+) 109 patients,group wound blush (-) 64 cases,the age,proportion of male patients,smoking history,diabetes,coronary heart disease,chronic renal insufficiency,pre and post operative ankle brachial index,were no statistical difference between the two groups.The ulcer healing time of group wound blush (+) was significantly shorter than that of group wound blush(-) (P ＜ 0.05).The rate of ulcer healing in group wound blush(+) was significantly higher than that in group wound blush(-) (P ＜ 0.05).In group wound blush(+),the cumulative rate of limb salvage was statisticallyhigher than group wound blush (-) (P ＜ 0.05).By logistic regression analysis,wound blush(-) (OR =4.5,P ＜ 0.05),IRc revascularization (OR =2.6,P ＜ 0.05) were independent risk factors of ulcer healing.Conclusions The resoult of wound blush(+) shows a good distal perfusion of foot.It can be used as a predictive factor for critical limb ischemia ischemic ulcer healing,and wound blush (-) was an independent risk factor for ulcer nonhealing.

Objective To study the relationship between single nucleotide polymorphisms of MMP1 gene (-1607) and cyclosporine-induced gingival overgrowth ( CsA-GO ) . Methods Polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP) were used to detect 42 cases of cyclosporine-induced gingival hyperplasia patients and 118 normal controls. MMP1 (-1607) 1G/2G gene polymorphism type were detected and analysised;Logistic regres-sion model was used to calculate the different genotypes and the risk of cyclosporine-induced gingival hyperplasia re-lationship. Results Cyclosporine-induced MMP1 (-1607) 3 genotype frequency distribution of gingival hyperplasiacase group had no significant difference (P=0.37) with the control group. Patients who carry MMP1 (-1607) 2G genotype occurred cyclosporine-induced gingival hyperplasia have 1.37 times risk than 1G genotype (95%CI=0.81-2.36, P=0.24), and 1G genotype and cyclosporine-inducedgingival hyperplasiaun related torisk. Conclusion MMP1 gene promoter region (-1607) 1G/2G single nucleotide polymorphism of cyclosporine -induced gingival hyperplasia may not a genetic susceptibility factor, patients who carry 2G genotype has an increased risk of CsA-GO.

Objective To compare the one year efficacy of intravitreal injection with ranibizumb for macular edema (ME) secondary to ischemic and non-ischemic central retinal vein occlusion (CRVO).Methods A total of 88 patients (88 eyes) with ME secondary to CRVO were enrolled in this retrospective study.The best corrected visual acuity (BCVA) was detected by the Early Treatment Diabetic Retinopathy Study Chart.The optical coherence tomography was used to measure the foveal retinal thickness (CRT) and macular edema volume.The patients were divided into non-ischemic group and ischemic group,44 eyes of 44 patients in each group.There was no significant differences in age (t=0.650,P=0.517) and gender (x2=0.436,P=0.509) between the two groups.Compared with the ischemic group,the CRT was significantly decreased in the non-ischemic group (t=-2.291,P=0.024),and the edema volume in the macular area was significantly reduced (t=-2.342,P=0.022).All eyes were treated with continuous intravitreal injection of ranibizumab three times,and repeated injections were performed as needed.The patients without obvious ME regression after treatment were combined with triamcinolone acetonide injection.The patients with peripheral retinal non-perfusion area were combined with peripheral retinal laser photocoagulation.The follow-up was 1 year.The number of injections was counted.The changes of BCVA,CRT and edema volume in the macular area were compared between the two groups.Results During the 1-year follow-up period,88 eyes were injected 1 to 10 times,with the mean of 4.51 ±2.33.The number of injections in the ischemic group and non-ischemic group were 4.55± 1.59 and 4.48 ± 2.91,respectively.There was no significant difference in the average number of injections between the two groups (t=0.136,P=0.892).The number of acetonide injections and laser treatment in the ischemic group was significantly higher than that in the non-ischemic group (t=3.729,9.512;P＜0.001).At the last follow-up,compared with the ischemic group,the BCVA was increased (t=8.128),the CRT was decreased (t=-7.029) and the edema volume in the macular area was decreased (t=-7.213) in the non-ischemic group (P＜ 0.001).Conclusion Compared with ME secondary to ischemic CRVO,intravitreal injection of ranibizumab for ME secondary to non-ischemic CRVO has the better outcome of vision improvement and edema regression as well as less fiequent of acetonide injections and laser treatment.

Objective:To explore the oxidative stress of cerebral white matter lesion (WML) and normal-appearing white matter (NAWM) with in vivo proton exchange rate (k ex) MRI on relapse-remitting multiple sclerosis (RRMS) patients. Methods:Clinical and imaging data of 37 patients (case group) with RRMS patients of Tongji Hospital of Tongji Medical College of Huazhong University of Science and Technology were analyzed retrospectively from November 2018 to November 2021, including 11 males and 26 females aged 18-41 (29±7) years. Another 22 age-matched healthy volunteers (control group) were recruited for the same period, including 4 males and 18 females aged 23-44 years with a median age of 25 (24, 28.25) years. All subjects received conventional MR protocols and chemical exchange saturation transfer imaging. The manifestation of WML on the k ex map and T 1WI images were assessed while the k ex values of WML, NAWM and normal white matter (NWM) of control group were quantitatively evaluated. Student′s t test was used to compare the k ex difference of WML and NAWM in the case group, NAWM in the case group and NWM in the control group, low-signal and isosignal WML in T 1WI. Spearman rank correlation was used to analyze the correlation of the k ex values of WML with patients′ expanded disability status scale (EDSS) score. Results:A total of 272 WML were found in the 37 RRMS patients, and 25.4% (69/272) were T 1-hypointense. The k ex value of WML in the case group [(932±108) s -1] was higher than that of NAWM [(771±26) s -1], and the difference was statistically significant ( t=8.95, P<0.001); the k ex value of NAWM in the case group [(771±26) s -1 ] was higher than that of NWM [(745±26) s -1] in the control group, and the difference was statistically significant ( t=3.96, P<0.001). The k ex value [(1 039±110) s -1] of WML with low signal at T 1WI was higher than that of WML with equal signal [(895±79) s -1], with a statistically significant difference ( t=9.78, P<0.001). Correlation analysis showed that the k ex value of WML in the case group was positively correlated with the EDSS score ( r=0.54, P<0.001). Conclusions:The elevated k ex values of WML and NAWM reflect the cerebral oxidative stress of RRMS patients and are positively correlated to the severity of tissue damage, which suggests the role of oxidative stress in RRMS lesion formation and brain atrophy.

Objective To investigate the effects of the third-generation mechanical chest compressor on outcomes of cardiopulmonary resuscitation in patients with out-of-hospital cardiac arrest.Methods The patients with out-of-hospital cardiac arrest from October 2015 to September 2017 in the Yuyao peoples' hospital were included and divided randomly into 2 groups:manual chest compression group and mechanical chest compression group.The duration of resuscitation,the rate of restoration of spontaneous circulation (ROSC),4-h survival rate and the rate of survival getting to hospital discharge with favorable neurological status of two groups were analyzed.Results A total of 95 patients with out-of-hospital cardiac arrest were included.The rate of ROSC and 4-h survival was significantly increased in the mechanical chest compression group compared with the manual chest compression group (P＜0.05).There was no remarkable difference in the duration of resuscitation and the rate of survival getting to hospital discharge with favorable neurological status between 2 groups.Conclusions The thirdgeneration mechanical chest compressor significantly improves the rate of ROSC and the shout-term survival in patients with out-of-hospital cardiac arrest.

Yeast autolysis affects the flavor and quality of beer. The regulation of yeast autolysis is a need for industrial beer production. Previous studies on brewer's yeast autolysis showed that the citric acid cycle-related genes had a great influence on yeast autolysis. To explore the contribution of isocitrate dehydrogenase genes in autolysis, the IDP1 and IDP2 genes were destroyed or overexpressed in typical lager yeast Pilsner. The destruction of IDP1 gene improved the anti-autolytic ability of yeast, and the anti-autolytic index after 96 h autolysis was 8.40, 1.5 times higher than that of the original strain. The destruction of IDP1 gene increased the supply of nicotinamide adenine dinucleotide phosphate (NADPH) and the NADPH/NADP⁺ ratio was 1.94. After fermentation, intracellular ATP level was 1.8 times higher than that of the original strain, while reactive oxygen species (ROS) was reduced by 10%. The destruction of IDP2 gene resulted in rapid autolysis and a decrease in the supply of NADPH. Anti-autolytic index after 96 h autolysis was 4.03 and the NADPH/NADP⁺ ratio was 0.89. After fermentation, intracellular ATP level was reduced by 8% compared with original strain, ROS was 1.3 times higher than that of the original strain. The results may help understand the regulation mechanism of citric acid cycle-related genes on yeast autolysis and provide a basis for the selection of excellent yeast with controllable anti-autolytic performance.
Humans ; Isocitrate Dehydrogenase/genetics* ; NADP ; Reactive Oxygen Species ; Autolysis ; Adenosine Triphosphate

The prefrontal cortex and hippocampus may support sequential working memory beyond episodic memory and spatial navigation. This stereoelectroencephalography (SEEG) study investigated how the dorsolateral prefrontal cortex (DLPFC) interacts with the hippocampus in the online processing of sequential information. Twenty patients with epilepsy (eight women, age 27.6 ± 8.2 years) completed a line ordering task with SEEG recordings over the DLPFC and the hippocampus. Participants showed longer thinking times and more recall errors when asked to arrange random lines clockwise (random trials) than to maintain ordered lines (ordered trials) before recalling the orientation of a particular line. First, the ordering-related increase in thinking time and recall error was associated with a transient theta power increase in the hippocampus and a sustained theta power increase in the DLPFC (3-10 Hz). In particular, the hippocampal theta power increase correlated with the memory precision of line orientation. Second, theta phase coherences between the DLPFC and hippocampus were enhanced for ordering, especially for more precisely memorized lines. Third, the theta band DLPFC → hippocampus influence was selectively enhanced for ordering, especially for more precisely memorized lines. This study suggests that theta oscillations may support DLPFC-hippocampal interactions in the online processing of sequential information.
Adult ; Female ; Humans ; Young Adult ; Epilepsy ; Hippocampus ; Memory, Short-Term ; Mental Recall ; Prefrontal Cortex ; Theta Rhythm ; Male