Objective To prepare virus-like particles (VLPs) for yellow fever virus (YFV) using the pACYCDuet-1 single-plasmid dual expression system. Methods Gene fragments including NS1, NS5, and 5' untranslated region (UTR) and 3' UTR fragment of YFV were cloned into the expression plasmid pACYCDuet-1-MS2-TMpac (carrying MS2 coat protein gene) and transformed into E. coli BL21(DE3) for expression. VLPs were observed by TEM. After DNase I/RNase A digestion, RNA was extracted and transcribed into cDNA, which was subjected to fluorescence quantitative PCR detection. Results According to SN/T 3560-2013, double digestion and colony PCR of pACYCDuet-1-MS2-TMpac-YFV both produced expected bands. SDS-PAGE showed a target band at 25-35 kDa. RT-PCR amplified VLPs after enzyme digestion, and TEM revealed VLPs with clear edges and translucent interiors. Thus, the plasmid was successfully constructed, and the phage coat protein packaged YFV genes with nuclease resistance. Conclusion VLPs containing specific gene fragments of yellow fever virus have been successfully prepared. These nuclease-resistant VLPs can serve as safe and stable quality control materials to ensure the reliability of laboratory detection for YFV.

Acute pancreatitis(AP)is a common and severe digestive disease in dogs,characterized by a high recurrence rate and complications that significantly impact canine health.This study in-vestigates the mechanism by which adipose-derived stem cells(ADSCs)regulate the CHOP/Bcl-2 pathway to mitigate acute pancreatic injury in dogs.Twenty Beagle dogs were randomly divided in-to four groups:a Sham group,an AP model group,an ADSCs treatment group,and a conditioned medium(CM)treatment group.The AP model was established by injecting a mixed solution of trypsin and sodium taurocholate into the AP model group.The ADSCs treatment group received intravenous injections of ADSCs immediately following surgery,while the CM treatment group re-ceived conditioned medium preparations.Pancreatic tissue was collected 24 hours post-surgery,and changes in the CHOP/Bcl-2 pathway were assessed using histopathology,transmission electron microscopy,western blotting,fluorescence quantification,immunofluorescence,and TUNEL stai-ning.The results indicated that AP induced significant interstitial edema,hemorrhage,inflammato-ry cell infiltration,chromatin contraction,and endoplasmic reticulum swelling,the expression lev-els of GRP78 and CHOP were found to be elevated,whereas the expression of Bcl-2 was downreg-ulated.Additionally,the expression levels of BAX,JNK,Caspase-3,and Caspase-12 increased,leading to an increased rate of cell apoptosis.Such changes result in an elevated apoptosis rate and a further decrease in Bcl-2 expression.Both ADSCs and CM therapy were found to alleviate the path-ological damage in pancreatic tissue,resulting in downregulation of CHOP and apoptosis-related markers,upregulation of Bcl-2,and a reduction in the apoptosis rate.The results of this study sug-gest that ADSCs may mitigate acute pancreatic injury induced by trypsin and sodium taurocholate in AP dogs by modulating the CHOP/Bcl-2 pathway.Targeted modulation of the transcriptional activity of CHOP may offer novel therapeutic approaches for AP.

Acute pancreatitis(AP)is a common and severe digestive disease in dogs,characterized by a high recurrence rate and complications that significantly impact canine health.This study in-vestigates the mechanism by which adipose-derived stem cells(ADSCs)regulate the CHOP/Bcl-2 pathway to mitigate acute pancreatic injury in dogs.Twenty Beagle dogs were randomly divided in-to four groups:a Sham group,an AP model group,an ADSCs treatment group,and a conditioned medium(CM)treatment group.The AP model was established by injecting a mixed solution of trypsin and sodium taurocholate into the AP model group.The ADSCs treatment group received intravenous injections of ADSCs immediately following surgery,while the CM treatment group re-ceived conditioned medium preparations.Pancreatic tissue was collected 24 hours post-surgery,and changes in the CHOP/Bcl-2 pathway were assessed using histopathology,transmission electron microscopy,western blotting,fluorescence quantification,immunofluorescence,and TUNEL stai-ning.The results indicated that AP induced significant interstitial edema,hemorrhage,inflammato-ry cell infiltration,chromatin contraction,and endoplasmic reticulum swelling,the expression lev-els of GRP78 and CHOP were found to be elevated,whereas the expression of Bcl-2 was downreg-ulated.Additionally,the expression levels of BAX,JNK,Caspase-3,and Caspase-12 increased,leading to an increased rate of cell apoptosis.Such changes result in an elevated apoptosis rate and a further decrease in Bcl-2 expression.Both ADSCs and CM therapy were found to alleviate the path-ological damage in pancreatic tissue,resulting in downregulation of CHOP and apoptosis-related markers,upregulation of Bcl-2,and a reduction in the apoptosis rate.The results of this study sug-gest that ADSCs may mitigate acute pancreatic injury induced by trypsin and sodium taurocholate in AP dogs by modulating the CHOP/Bcl-2 pathway.Targeted modulation of the transcriptional activity of CHOP may offer novel therapeutic approaches for AP.

Pulmonary hypertension has a high mortality rate,and although targeted therapy is available,it is still incurable,and the long-term prognosis for patients is poor.As a tyrosine kinase inhibitor,imatinib was approved for marketing in China in 2002 for the treatment of chronic myelogenous leukemia and other tumor diseases.In addition to the antitumor effects,imatinib was found to improve hemodynamics and exercise tolerance in patients with severe pulmonary arterial hypertension,but the safety was suboptimal.With the emergence of new formulations of imatinib targeted at the lungs,it is expected to become a new targeted drug for pulmonary arterial hypertension.

Microneedle percutaneous immunization is achieved by puncturing the stratum corneum of the skin with microneedles so that the vaccine is efficiently recognized by antigen-presenting cells to induce a specific immune response. Due to the advantages of efficient induction of immune response, low pain and easy storage, transdermal immunization by microneedles has been widely used for immunization of various vaccines in recent years. This review summarizes the materials of microneedles, application for transcutaneous immunization, as well as the challenges that need to be addressed.
Administration, Cutaneous ; Drug Delivery Systems ; Needles ; Vaccination ; Vaccines

Objective:To investigate the effect of free anterolateral thigh perforator flap in repair of forefoot injuries combined with multiple tissue defect.Methods:A retrospective case series study was conducted on 26 patients who suffered from forefoot injuries combined with multiple tissue defect admitted to 920th Hospital of Joint Logistic Support Force of PLA from January 2015 to December 2019. There were 21 males and 5 females, aged 15-61 years [(31.6±12.5)years]. The combined injuries were fracture in 10 patients, arsometatarsal joint dislocation in 3, bone defect in 9, tendon injury in 5, and ligament injury in 3. Management of multiple tissue defect of the forefeet: soft-tissue defect of the forefeet was resurfaced with free anterolateral thigh perforator flaps with the dimension of 6.0 cm×3.5 cm to 26.5 cm×10.0 cm; fracture was fixed by Kirschner wires; joint dislocation was treated by open reduction and Kirschner wires fixations; bone defect was reconstructed either by one-stage bone graft or by use of membrane-induced technique and secondary bone graft, according to the wound conditions; tendon injury of extensor digitorum longus was repaired by direct tendon suture or by tendon transfer; tarsometatarsal ligament injury was primarily sutured. The flap survival rate was observed within 2 weeks after operation. The fracture healing, bone-defect repair, foot appearance, and donor-site healing were detected at 1 month, 3 months, 6 months, 1 year post-operatively and at the last follow-up. The postoperative complications were recorded. The foot function was assessed using American Orthopedic Foot and Ankle Society (AOFAS) ankle-hindfoot score before operation and at the final follow-up.Results:All patients were followed up for 6-36 months [(20.5±4.6)months]. All flaps survived uneventfully. The fracture healing and bone defect repair were acquired. The flap showed good texture, including primary flap thinning in 11 patients and secondarily thinning in 15 patients at 3-6 months postoperatively. The donor sites showed good healing, leaving only a linear scar. The flap venous crisis developed in 1 patient and survived after emergency vascular exploring. Local infection of flap occurred in 3 patients and was cured after further debridement and the use of sensitive antibiotics. The AOFAS ankle-hindfoot score was 54-94 points [(76.6±10.4)points] at the last follow-up, compared to preoperative 11-51 points [(27.2±11.3)points] ( P<0.01). The results were excellent in 5 patients, good in 11, and fair in 10, with the excellent and good rate of 62%. Conclusions:For forefoot injuries combined with multiple tissue defect, anterolateral thigh perforator flap transplantation with additional techniques to treat fractures, bone defect, tendon and ligament injuries can achieve satisfactory results in aesthetic appearance of the flap and donor site and foot function recovery.

Objective: To investigate the metabolic patterns of ¹¹C-2-beta-carbomethoxy-3-beta-(4-fluorophenyl)tropane (CFT) and ¹⁸F-fluorodeoxyglucose (FDG) PET/CT imaging in patients with tremor and non-tremor Parkinson′s disease (PD). Methods: From March 2018 to March 2019, 28 PD patients (19 tremor patients: 14 males and 5 females, age: (59.9±11.4) years; 9 non-tremor patients: 3 males and 6 females, age: (62.6±9.0) years) were enrolled. For the two groups, the ¹¹C-CFT uptake values in caudate nucleus, anterior putamen and posterior putamen as well as ¹⁸F-FDG uptake values in all brain regions were calculated by regions of interest (ROI) method. Two-sample t test or Mann-Whitney u test were used to analyze the data. Results: Caudate nucleus ¹¹C-CFT uptake in PD patients with tremor was higher than that without tremor (3.03±0.51 vs 2.60±0.62; t=2.687, P<0.05). Yet thalamus glucose uptake of non-tremor PD patients showed more active than tremor patients (1.14±0.05 vs 1.10±0.03; t=3.449, P<0.01). Sorted by modified Hoehn-Yahr stage, tremor patients in advanced stage (stage≥2.5) were characterized by glucose uptake increase in the putamen and cerebellum compared with non-tremor advanced group (1.27±0.04 vs 1.21±0.05, 0.94±0.04 vs 0.86±0.08; t values: 2.695 and 2.492, both P<0.05). Metabolic decrease in the premotor areas was observed in tremor patients in advanced stage compared with early stage (1.07±0.02 vs 1.10±0.03; t=2.053, P<0.05). Conclusion ¹¹C-CFT and ¹⁸F-FDG PET/CT imaging can indicate different metabolic patterns between tremor and non-tremor PD, thus providing information for clinical practice and differential diagnosis.

Objective To explore the clinical efficacy of Ilizarov technique for infected-bone defect in lower extremity.Methods From January 2013 to December 2016,data of 31 consecutive patients with lower limb infected bone defect were retrospectively analyzed.There were 23 males and 8 females,with an average age of 35.8 years (range,5-57 years).16 cases of infected bone defect were on femur while the other 15 were on tibia.The infected bone defects were developed from blood-borne osteomyelitis in 3 cases,from open fracture (according to Gustlio classification,Gustilo Ⅱ 8,Ⅲ a 5,Ⅲ b 4) surgery in 17 cases (12 cases after external fixation and 5 cases after internal fixation),and from closed fracture surgery of plate or intramedullary nail fixation in 11 cases.Soft-tissue defects were observed in 5 cases,including 1 femur and 4 tibia.The previous operations were performed with an average operation time of 3.2 (range,2-5 times) and treatment duration of 9.6 months (range,3-21 months).The operation process was one stage debridement,external fixation,combined with bone osteotomy and transportation.Bone transportation started 7 to 10 days after osteotomy with a speed of 1 mm/d,and 0.25 mm/time.Healing rate and complications were recorded during the follow-up period and were assessed by Paley fracture healing score.Results The length of bone defect after debridement was 6.9 cm (range,3-13.5 cm).There were 27 cases positive in bacterial culture,who were treated by sensitive antibiotics.The other 4 cases were negative and treated by broad-spectrum antibiotics.The bone lengthening speed was 0.76 mm/d (range,0.56-0.86 mm/d).All bone and soft-tissue defects healed,with average bone healing time of 11.5 months (range,6-18.5 months).All patients were followed-up for 12 to 48 months (average,25.5 months).Pin infection appeared in 11 cases (8 cases on femur and 3 cases on tibia)who were cured by oral antibiotics,serial dressing and debridement.Axial displacement was presented in 4 cases on femur and were corrected by external fixation adjustment.No recurrent infection was observed and the limb length restored normal.Postoperative complications in femur were 106.3％ (17/16) comparing 46.7％ (7/15) in tibia.According to Paley fracture healing score,12 cases were excellent,14 were good and 5 were fair,yielding an excellent to good rate of 83.9％ (26/31).Conclusion Ilizarov technique is an effective way in the treatment of infected-bone defects.The complication rates on femur observed in this study were slightly higher than that on tibia.

Objective:To investigate the mainstream view and statusof people on body donation in Yunnan province, and provide references. Methods:Random sampling questionnaire survey was used, the obtained data was statistically analyzed. Results:Only 48. 0% of the people understand the body donation, 20. 6% of people know body donation ways and procedures, 27. 8% of people have heard of body receiving station, 11. 4% of people know the body receiving station in yunnan province, 17. 2% of the people to understand body donation of relevant laws and regulations. 47. 4% of people that are willing to donate his body. Conclusion:Make perfect laws and reg-ulations, respect for the body, giving donors and their families allowance, body donation monument, to carry out education and publicity of the body donation form to the whole society support and participate in the good atmos-phere of body donation has great benefits.

OBJECTIVETo explore the impacts on the life quality and the effect mechanism in the patients of allergic rhinitis (AR) treated with warm acupuncture in winter and summer.

METHODSTwo hundred and forty patients of AR were randomized into a summer and winter acupuncture group, a non-summer and winter acupuncture group and a western medication group, 80 cases in each one. In the two acupuncture groups, Dazhui (GV 14), Fengmen (BL 12), Feishu (BL 13), Pishu (BL 20) and Shenshu (BL 23) were selected. In the summer and winter acupuncture group, the warm acupuncture started at the first day of the three periods of hot season and the first day of the third nine-day period after the winter solstice. The treatment was given once every two days, continuously for 15 times. Totally, 30 treatments were required a year. In the non-summer and winter acupuncture group, the warm acupuncture was applied out of the three periods of the hot season and the third nine-day period after the winter solstice. The treatment was given once every two days and 30 treatments for a year. In the western medication group, cetirizine was taken orally, continuously for 30 days as one session. In the three groups, the treatment for 1 year was taken as one session. The second session started in the next year. Totally, 2 sessions were required. The score of rhinoconjunctivitis quality of life questionnaire (RQLQ) and the level of serum immunoglobulin E (IgE) were compared in the patients' of each group before treatment and in 1 and 2 sessions of treatment.

RESULTSAfter treatment, the scores of 7 domains, named activities, common complaints, practical problems, sleep, ocular symptoms, nasal symptoms and emotions were all improved as compared with those before treatment, in the patients of the three groups (all P < 0.05). After 2 sessions treatment, the results in the summer and winter acupuncture group were better than those in the other two groups (all P < 0.05), and the results in the non-summer and winter acupuncture group were better than those in the western medication group (all P < 0.05). After treatment, serum IgE level was lower averagely than that before treatment in the three groups (all P < 0.05). After 2 sessions treatment, the result in the summer and winter acupuncture group was lower than that in the other two groups (both P < 0.05), and the result in the non-summer and winter acupuncture group was lower than that in the western medication group (P < 0.05). After treatment, the total effective rate was 92.3% (72/78) in the summer and winter acupuncture group, better than 80.5% (62/77) in the non-summer and winter acupuncture group and 69.7% (53/76) in the western medication group (both P < 0.05).

CONCLUSIONThe warm acupuncture in both summer and winter achieves the definite efficacy on AR and the effect mechanism is relevant with reducing serum IgE level.

Acupuncture Points ; Acupuncture Therapy ; instrumentation ; methods ; Adolescent ; Adult ; Female ; Humans ; Male ; Middle Aged ; Quality of Life ; Rhinitis, Allergic ; therapy ; Seasons ; Treatment Outcome ; Young Adult