Objective:To develop a recombinant protein vaccine based on KPC-2, a drug resistance target in Klebsiella pneumoniae, and evaluate its immunogenicity, protective efficacy and mechanism in a mouse model of pneumonia. Methods:KPC-2 was expressed in Escherichia coli and purified using GST affinity chromatography. A recombinant protein vaccine was prepared with KPC-2 and used to immunize New Zealand rabbits through subcutaneous injection. Serum samples were isolated from cardiac blood and Protein G chromatography was used to purify polyclonal antibodies against KPC-2. Opsonophagocytic killing assay was used to assess the bactericidal activity of the polyclonal antibodies in vitro. Female BALB/c mice were immunized three times with the recombinant protein vaccine, and the titers of specific IgG antibodies in serum were measured by indirect ELISA. One week after the last vaccination, the mice were infected with Klebsiella pneumoniae strain SRT through tracheal intubation, and received a single intravenous dose of meropenem (0.1 mg) 1 h later. The protective efficacy of the KPC-2 recombinant protein vaccine was evaluated by comparing the survival rates, bacterial colonization and histopathological changes between vaccine group and adjuvant group as well as the survival rates between meropenem group and normal saline group. Moreover, the protective efficacy of polyclonal antibodies against KPC-2 was evaluated through passive immunization. Results:The level of specific IgG antibodies in serum was significantly higher in the vaccine group than in the adjuvant group ( t=4.325, P<0.05). The survival rate in the vaccine group was also higher than that of the adjuvant group [70% (7/10) vs 10% (1/10), P<0.05]. Furthermore, lung inflammation was less severe and bacterial burden was reduced in the vaccine group as compared with those of the control group ( t=3.127, P<0.05). Both active and passive vaccination strategies demonstrated strong protective efficacy against Klebsiella pneumoniae infection, and had a synergistic effect when used in combination with antibiotic therapy. The polyclonal antibodies against KPC-2 had bactericidal activity in vitro ( t=5.427, P<0.05). Conclusions:The prepared KPC-2 vaccine has better immunogenicity and protective efficacy. It can induce strong humoral immune responses. This study suggest that drug resistance target may be used as a candidate antigen for future vaccine development.

Objective To investigate the effect of altered oxidative stress system on liver function after partial splenic embolization(PSE).Methods Twenty-nine patients with liver cirrhosis and hypersplenism who received PSE were selected.Peripheral venous blood was drawn from the patients before and at 1 week,1 month,and 3 months after PSE,and the indexes of oxidative stress system factors including malondialdehyde(MDA),superoxide dismutase(SOD),advanced oxidiation protein products(AOPPs),and gluta-thione peroxidase(GSH-Px)were detected,as well as liver function indexes.Results There were positive correlation between SOD activity and total bilirubin(TBil)and model for end-stage liver disease(MELD)scores at 1 week postoperatively(TBil:r=0.725,P＜0.05;MELD:r=0.764,P＜0.05).There was positive correlation between GSH-Px activity and alanine aminotransferase(ALT)at 1 month postoperatively(r=0.777,P＜0.05),however,the AOPPs was negatively correlated with ALT and aspartate aminotransferase(AST)at 3 months postoperatively(ALT:r--0.900,P＜0.05;AST:r=-0.957,P＜0.05).Conclusion PSE can improve the body oxidative stress system and enhance the body's antioxidant response,and then improve the liver function.

In vitro studies have established the prevalent theory that the mitochondrial kinase PINK1 protects neurodegeneration by removing damaged mitochondria in Parkinson's disease (PD). However, difficulty in detecting endogenous PINK1 protein in rodent brains and cell lines has prevented the rigorous investigation of the in vivo role of PINK1. Here we report that PINK1 kinase form is selectively expressed in the human and monkey brains. CRISPR/Cas9-mediated deficiency of PINK1 causes similar neurodegeneration in the brains of fetal and adult monkeys as well as cultured monkey neurons without affecting mitochondrial protein expression and morphology. Importantly, PINK1 mutations in the primate brain and human cells reduce protein phosphorylation that is important for neuronal function and survival. Our findings suggest that PINK1 kinase activity rather than its mitochondrial function is essential for the neuronal survival in the primate brains and that its kinase dysfunction could be involved in the pathogenesis of PD.

Objective:To automatically synthesize Al 18F-1, 4, 7-triazacyclononane-1, 4, 7-triacetic acid (NOTA)-fibroblast activation protein inhibitor (FAPI)-04, perform PET/CT imaging in vivo, and evaluate its diagnostic efficacy on tumors. Methods:Al 18F-NOTA-FAPI-04 was produced in All-in-one automatic synthesis module and its quality control was conducted by high performance liquid chromatography (HPLC) equipped with a radioactive detector. Al 18F-NOTA-FAPI-04 PET/CT imaging was performed in normal BALB/c mice ( n=3) and 4T1 breast cancer models ( n=3) to determine its biodistribution. Then Al 18F-NOTA-FAPI-04 and 18F-FDG PET/CT imaging were performed in a hepatocellular carcinoma patient (male, 51 years old). Results:The synthesis time of Al 18F-NOTA-FAPI-04 was about 35 min, and the radiochemical yield was (25.2±1.9)% (attenuation correction, n=3). The product was colorless transparent solution with pH value of 7.0-7.5, and the specific activity was (46.11±3.07) GBq/μmol (attenuation correction, n=3). The radiochemical purity was above 99.0% and was still above 98.0% at room temperature after 6 h. PET/CT imaging in mice showed that physiological uptake of Al 18F-NOTA-FAPI-04 was mainly in biliary system and bladder, and Al 18F-NOTA-FAPI-04 highly concentrated in tumor xenografts. PET/CT imaging in the patient showed that Al 18F-NOTA-FAPI-04 obtained high tumor background ratio (TBR) values of 4.1, 8.9, 5.4, 4.8, 2.2 in parasternal lymph nodes, anterior diaphragmatic lymph nodes, hilar lymph nodes, pancreaticoduodenal ligament lymph nodes, abdominal aortic lymph nodes, respectively, while TBR values were 1.0, 2.8, 5.0, 2.1, 1.1 by 18F-FDG. Conclusions:Al 18F-NOTA-FAPI-04 can be synthesized with short time, high radiochemical yield and good stability using All-in-one module. Al 18F-NOTA-FAPI-04 PET/CT imaging has high contrast and excellent diagnostic efficacy on tumors.

Objective:To investigate the evaluation value of one-stop whole-brain dynamic volume CT angiography and CT perfusion imaging (CTA-CTP) in the cynomolgus monkeys models of middle cerebral artery occlusion (MCAO).Methods:Ten adult cynomolgus monkeys were selected and examined by head and neck CTA-CTP and craniocerebral MRI to rule out craniocerebral space-occupying lesions or cerebrovascular malformation. Under guidance of digital substraction angiography (DSA), the right femoral artery was dissected and monkey autologous thrombosis was injected into the right middle cerebral artery (MCA) through microcatheter to prepare MCAO models. Whole brain DSA was performed intraoperatively to observe whether the model was successfully prepared, and head and neck CTA-CTP was performed 24 h and 7 d after modeling to determine the locations and brain blood flow changes of ischemic lesions. The monkeys were sacrificed 8 d after modeling, and the brain tissues were stained with 2,3,5-triphenyltetrazolium chloride (TTC).Results:Among the 10 cynomolgus monkeys, one was excluded because of preoperative cerebrovascular malformation, and one died of cerebral hernia caused by cerebral hemorrhage during the experiment. The remaining 8 MCAO models were successfully prepared. Intraoperative DSA orthography showed unclear M1 segment and distal branch of MCA. Brain CT scan 24 h and 7 d after modeling showed obvious cerebral ischemic lesions in the right MCA blood supply area, and the infarct extent 7 d after surgery was more obvious than that 24 h after surgery. CTA examination showed obvious blood flow interruption imaging in the in M1 segment of MCA on the right side, the distal vessels were not clearly displayed and the distal branches of the infarct side 7 d after surgery were obvious decreased as compared with those 24 h after surgery. CTP scan showed that the cerebral blood volume of the right cerebrum was obviously reduced as compared with that of the left cerebrum, which was consistent with the blood supply area of MCA; and the infarct cores and penumbra areas 7 d after surgery were obvious increased as compared with those 24 h after surgery. TTC staining showed that the ischemic lesions of the brain tissue on the slices were gray and involved multiple layers, and the range was roughly consistent with the infarction sites shown by DSA and CT imaging.Conclusion:One-stop whole brain dynamic volume CTA-CTP has good evaluation value in imaging findings in MCAO animal models.

Humans ; Male ; Middle Aged ; Paranasal Sinuses ; diagnostic imaging ; Sublingual Gland Neoplasms ; diagnosis

Objective:To establish the quality standard for Aidi B capsules.Methods:Astragalus membranaceus,Fallopia multi-flora and Gastrodiae elata were identified by TLC qualitatively.The content of gastrodin was determined by HPLC.The chromatographic separation was carried out on an Eclipse Plus C18column (250 mm ×4.6 mm,5 μm). The mobile phase consisted of acetonitrile-0.05% phosphoric acid solution (2:98) at a flow rate of 1.0 ml·min-1.The detection wavelength and the column temperature was 220 nm and 25℃,respectively.Results:The spots in TLC were clear without any interference.The linear range of gastrodin was 8.532-208.8 μg·ml-1(r=1.000 0),and the average recovery was 100.30%(RSD=0.37%,n=6).Conclusion:The method is simple with good repeatability,which can be used for the quality control of Aidi B capsules.

Objective To evaluate the accuracy and feasibility of primary total knee arthroplasty (TKA) assisted by the self-made extramedullary osteotomy template on distal femur alignment.Methods We retrospectively analyzed the clinical data of 40 patients with osteoarthritis or rheumatoid undergoing primary TKA from September 2015 to March 2016.All patients were divided into two groups,extramedullary group and intramedullary group.Extramedullary group had 20 patients (4 males and 16 females) aged 65.1±6.9 years (range 51-75).Intramedullary group had 20 patients (3 males and 17 females) aged 65.7±7.9 years (range 48-75).The blood loss volume,drainage in operation,postoperative range of motion (ROM) at 1 week and 2 weeks,deviation between the ideal value and preoperative hip-knee-ankle (HKA) angle,coronal and sagittal angle of the femoral prosthesis were compared within the two groups.Results The age,gender,body mass index,deviation between the ideal value and preoperative HKA angle,Hospital for Special Surgery (HSS) scoring system,ROM of knee were not significantly different between the two groups.The operation time was 97.4± 14.0 min in extramedullary group and 111.7±9.9 min in intramedullary group (P ＜ 0.05).The blood loss was 71.8± 14.0 ml in extramedullary group and 111.7± 10.0 ml in intramedullary group (P ＜ 0.05),while the drainage volume was 167.0±74.5 ml in extramedullary group and 243.2±91.6 ml in intramedullary group (P＜ 0.05).The postoperative mean deviation of the HKA angle in extramedullary group (2.37°±0.73°) was smaller than that in intramedullary group (3.26°±1.42°).Furthermore,the coronal alignment of the femoral prosthesis in extramedullary group (89.70°±1.91°) was larger than that in intramedullary group (87.69°±2.39°).The sagittal alignment of the femoral prosthesis in extramedullary group (90.03°±2.62°) was also significantly larger than that in intramedullary group (88.07°±2.43°,P ＜ 0.05).The postoperative ROM of knee at 1 and 2 week in the extramedullary group were 101.75°±7.48° and 114.00°±7.88° respectively,while those in the intramedullary group were 101.25°±8.41° and 114.25°±8.78°.There was no statistically different between the two groups.There was no intraoperative and postoperative early complications reported in the two groups.Conclusion Although the primary TKA assisted by the selfmade extramedullary osteotomy template on distal femur alignment can shorten operation time,decrease the blood loss and increase the accuracy of coronal and sagittal alignment,the HSS score and ROM of knee are similar as intramedullary method.

Objective:To establish the quality standard for Shule capsules. Methods: The microscopic characteristic identifica-tion of Figwort and Radix bupleuri was performed under a microscope. The qualitative identification of fritillary, andrographispaniculata and polygonumbistorta was studied by TLC. The content of salvianolic acid B in Salvia miltiorrhiza was determined by HPLC. The chro-matographic separation was carried out by using a phenomenex synergi 4 hydro-RP 80A (250 mm × 4. 6 mm) column. The mobile phase consisted of acetonitrile-methanol-formic acid-water(10 :30 :59 :1)with gradient elution at a flow rate of 1. 0 ml·min-1, and the injection volume was 10 μl . The detection wavelength and the column temperature was 286 nm and 20 ℃, respectively. Results:The microscopic characteristics were obvious. The spots in TLC were clear without any interference. The linear range was 10. 001-100. 007 μg·ml-1(r=1. 0000). The average recovery was 99. 3% with RSD of 1. 8% (n=6). Conclusion:The method is simple with high specificity and good repeatability, which can be used as the quality control method for Shule capsules.

Objective:To establish the quality standard for Shenyan Ⅱ granules. Methods:The characteristics of Folium Pyrrosiae were identified by microscopy. The three herbs in the preparation,rehmannia,madder and thistle were identified by TLC qualitatively. The content of ginsenoside in American ginseng was determined by HPLC. Chromatographic separation was carried out by using a WONDER CRACT ODS-2(150 mm × 46 mm,5 μm)column. The mobile phase consisted of acetonitrile-0. 1% phosphoric acid(28 ∶72)with gradient elution at a flow rate of 1. 0 ml·min -1 ,and the injection volume was 10 μl. The detection wavelength and the column temperature was 203 nm and 40 ℃,respectively. Results:The spots in TLC were clear without any interference. The linear range of ginsenoside was 38. 9- 194. 5 μg·ml -1(r = 0. 999 3). The average recovery was 98. 3% and RSD was 1. 9%(n = 6). Conclusion:The method is simple and highly specific with good repeatability,which can be used as the quality control method for Shenyan Ⅱ granules.