Objective An adeno associated virus (AAV)vector targeted to hepatoma cells was constructed in order to be used in the apoptosis inducing gene therapy of liver cancer. Methods Firstly, primers containing specific enzyme cutting sites was designed and used to amplify the alpha fetoprotein promoter(AFP promoter) from human genome; Secondly, the promoter was cloned into the eukyrocyte expressing plasmid pTR UF5,resulting in the recombinant AAV vector plasmid containing the reporter gene( rAAV AFP GFP ); thirdly, blunted ligation was applied to construct the recombinant AAV vector plasmid containing the p53 gene( rAAV AFP p53 ). Results Recombinant adeno associated virus plasmid was constructed successfully that carried human wild type p53 gene under the control of human AFP promoter. Conclusion Theoretically the recombinant adeno associated virus plasmid rAAV AFP p53 we constructed could be used in gene therapy for hepatocellular carcinoma.

Objective To explore the effect of Jisuikang on neural functional recovery, and expression of brain-derived neurotrophic factor (BDNF) protein and mRNA level after spinal cord injury (SCI). Methods 144 female Sprague-Dawley rats, weighted 180 to 220 g, were used for experiment. 24 rats were randomly extracted into sham group (Group A), which had their vertebral plates and spines bitten away only.The others were randomly divided into model group (Group B), prednison group (Group C), and high, middle and low doses of Jisuikang group (Groups D to F) after SCI, 24 rats in each group. Group C was given 0.06 g/(kg ⋅ d) prednison, and Groups D to F were given 50, 25 and 12.5 g/(kg ⋅d) Jisuikang respectively, which were given 20 ml/(kg ⋅d) volume by intragastric administration. Groups A and B were given the same volume of normal saline (NS). The Basso-Beattie-Bresnahan (BBB) scores and oblique board test were applied to test the postoperative results 24 hours, 3, 7 and 14 days after SCI. The rats were executed and the spinal cord tissues were extracted 3, 7 and 14 days after SCI. Immunohistochemistry, Western blotting and RQ-PCR were applied to test the expression of protein and mRNA of BDNF. Results BBB scores and angle of oblique board test were significantly lower in Groups B to F than in Group A 24 hours after SCI (P<0.01). BBB scores were higher in both Groups C and E than in Group B 3 to 14 days after SCI (P<0.05), and was significantly higher in Group E than in the other groups 14 days after SCI (P<0.01). The results of immunohistochemistry and Western blotting showed that the protein expression of BDNF were significantly higher in Groups C and E than in Group B at different time points in the injured area after SCI (P<0.01), while there was no significant difference between Groups C and E (P>0.05). The results of RQ-PCR showed that prednisone and Jisuikang promoted the expression of BDNF mRNA. Group C (prednisone) had a most obvious effect at the beginning while Group E was better than Group C 14 days after SCI. Conclusion Jisuikang can promote the neural functional recovery and the expression of BDNF on both protein and mRNA level in SCI rats.

PurposeTo evaluate multislice spiral CT (MSCT) in diagnosing abdominal anaphylactoid purpura (AAP), and to explore its role in treatment follow-up.Materials and Methods Clinical and MSCT data of 13 patients with confirmed AAP were retrospectively analyzed. All patients underwent abdominal CT scan and thin layer reconstruction. Upper endoscopy was performed in 8 patients prior to treatment. MSCT was performed in 10 patients and endoscopy in 4 patients posttreatment, then clinical and CT features were compared to pretreatment findings.Results In pretreatment CT scan, single segment bowel involvement was found in 2 cases, multisegmental bowel involvement in 10 cases, and no positive finding in 1 patient. The duodenum and jejunum were involved in 8 patients and stomach in 5 patients. The diseased bowel wall showed swelling and thickening with decreased attenuation and homogeneous luminal narrowing with equivocal lining and double loop sign. Infiltration was found in 10 patients, small ascites in 3 patients. Patients were misdiagnosed as acute pancreatitis in 2 cases, acute cholecystitis, small bowel obstruction and peritonitis in 1 case respectively. Of 13 patients, five patients were cured, and the other 8 patients were improved. The clinical symptoms including rash, abdominal pain, occult blood, leukocytosis, vomiting, melena, urine occult blood were improved (χ2=5.59-18.33,P<0.05 orP<0.01). Post treatment CT showed significant improvement of bowel edema and infiltration (P<0.01). There was no statistically significant difference in change of ascites (P>0.05).Conclusion MSCT findings of AAP are nonspecific. CT diagnosis is difficult before skin rash. Combining CT characteristics of multisegmental bowel edema and clinical manifestations is helpful. CT examination can effectively follow up treatment response.

BACKGROUND:Studies have shown that Clematis chinensis Osbeck can promote articular chondrocyte proliferation, maintain and promote the synthesis of cartilage proteoglycan and type II col agen in chondrocytes. Low-intensity pulsed ultrasound can effectively improve the carticular chondrocyte proliferation, increase the cellmembrane permeability, and promote chemicals or genes delivery, so as to improve the biological effects of chemicals.
OBJECTIVE:To observe the effect of low-intensity pulsed ultrasound mediated Clematis chinensis Osbeck on the proliferation of cartilage cells and the expression of type II col agen and transforming growth factor (TGF)-β1 of rabbit knee articular chondrocytes cultured in vitro, and explore the effect and mechanism of Clematis chinensis Osbeck mediated by low-intensity pulsed ultrasound on cartilage damage.
METHODThe rabbit knee articular chondrocytes were isolated and cultured in vitro. cells of the second generation in logarithmic growth phase were randomly divided into four groupcontrol group, low-intensity pulsed ultrasound (LIPUS) group, Clematis chinensis Osbeck (CCO) group, and CCO+LIPUS group. After cells were cultured under different conditions for 3 days, cellproliferation was measured with cellCount Kit-8 assay. The secretion of type II col agen was detected by cytochemical staining method, and the expression of TGF-β1 was assayed by western blot analysis.
RESULTS AND CONCLUSION:The number of cells in the other three groups were significantly higher than that in control group at 3 days after culture (P<0.05), while the CCO+LIPUS group had the most cells. Cytochemical staining showed that the area and absorbance value of type II col agen in cartilage cells were significantly increased in CCO+LIPUS group, compared with control group (P<0.05), and the difference with control group was larger than the amount of that of LIPUS group and CCO group with control group. Western blot analysis showed that TGF-β1 was significantly expressed in CCO+LIPUS group, and the relative gray value was significantly higher than the other groups (P<0.05). Both low-intensity pulsed ultrasound and Clematis chinensis Osbeck can promote the proliferation of cartilage cells and expression of type II col agen and t-β1 of rabbit knee articular chondrocytes cultured in vitro. The combined method could produce synergistic effects.

Objective:To investigate the epidemiological and clinical features of patients with hemorrhagic fever with renal syndrome (HFRS) from 2017 to 2019.Methods:Seventy-five patients with HFRS from the Department of Infectious Diseases, The First Affiliated Hospital of Anhui Medical University during January 1, 2017 to December 31, 2019 were included. The data of epidemiology, clinical symptoms, blood routine, urine routine, serum creatinine, liver function and other laboratory examination indexes were retrospectively analyzed. The measurement data with skewness distribution were expressed by M( QR) and compared by nonparametric test. Multivariate logistic regression analysis was used to analyze disease-related risk factors. Results:The 75 patients were mainly located in the western and northern regions of Anhui Province. A total of 37 cases (49.3%) were infected during November, December and January next year. Fifty-four (72.0%) patients were farmers and 10(13.3%) patients had a clear history of rodent contact. Only 19(25.3%) patients had typical clinical manifestations of "three red and three pain" . Fifty-eight (77.3%) patients had elevated white blood cell count, 67(89.3%) patients had decreased platelet count, 55(73.3%) patients had urinary protein + + + , 65(86.7%) patients had abnormal urinary occult blood, and 67(89.3%) patients had elevated serum creatinine. The serum creatinine and potassium levels in 31 severe and critical patients were 495(301) μmol/L and 4.14(0.77) mmol/L, respectively, which were both higher than those in 44 mild and moderate patients (235(289) μmol/L and 3.65(1.02) mmol/L, respectively). The differences were both statistically significant ( Z=-3.187 and -2.796, respectively, both P<0.01). Multivariate logistic regression analysis showed that serum creatinine (odds ratio ( OR)=1.005, 95% confidence interval ( CI)1.002-1.008) and serum potassium ( OR=2.632, 95% CI 1.098-6.313) were independent risk factors for disease severity. All patients received comprehensive medical treatment, and 27 patients received renal replacement therapy. Sixty-eight patients had good prognosis and four patients died. Conclusions:HFRS is still common in the rural area in winter and spring. Patients with atypical clinical manifestations and severe and critical patients should be intensively monitored.

Objective To establish a simple and sensitive method for the determination of plasma drug concentration in rats after intragastrical and intravenous administration of paclitaxel loaded nanoparticles and to evaluate its pharmacokinetic characteristics .Methods AgilentTC-C18 column (250 mm × 4 .6 mm ,5 μm) was used for HPLC at the flow rate 1 ml/min with ketoconazole as internal standard and methanol acetonitrile water (45:20:35) as the mobile phase .The statistical mo-ment method was applied to calculate the pharmacokinetic parameters and to evaluate pharmacokinetic characteristics .Results There was a good liner relationship (r=0 .9997) when rat blood concentration of Paclitaxel ranged from 0 .5 to 20μg /ml .The accuracy and precision were in line with the requirements of biological sample analysis .The t1/2 were 3.86 ,3.76 ,3.35 and 2 .62 h after intravenous injection of three lab-made paclitaxel nanoparticles and paclitaxel solution via rat tail vein .The t1/2 were 5 .28 and 3 .72 h respectively after intragastrical administration of lab-made paclitaxel nanoparticles and paclitaxel suspen-sion .Conclusion This method can be used for the pharmacokinetic study of paclitaxel nanoparticles in rats .The paclitaxel loaded nanoparticles exhibited significantly longer in vivo retention time than paclitaxel injection and paclitaxel suspension .

ObjectiveTo explore the expression of CD3+ CD8+ human leukocyte antigen (HLA)-A2+T lymphocytes with specificity to the different hepatitis B virus (HBV) peptides in the peripheral blood mononuclear cells (PBMC)from the patients with hepatitis B associated hepatocellular carcinoma (HCC).MethodsThe HLA-A2+ PBMC from four patients with hepatitis B associated HCC were incubated with five HBV/HLA-A2 pentamers respectively,which were HBV sAg (FLLTRILTI),HBV sAg (GLSPTVWLSV),HBV sAg (WLSLLVPFV),HBV core (FLPSDFFPSV),and HBV pol (FLLSLGIHL),as well as anti-CD3-pacific blue and anti-CD8-fluorescein isothiocyanate (FITC).Then,HBV/HLA-A2-CD3-CD8 positive cells were detected by flow cytometry. The monoclonal HBV/HLA-A2-CD3-CD8+ cells were acquired by fluorescenceactivated cell sorter,and cultured and identified by flow cytometry.The anti-HBV specific T lymphocytes were then cultured with HepG2 (HLA-A2+ ) cells and the release of interferon γ (IFN-γ)were determined by enzyme-linked immunosorbent assay (ELISA),Res(a)ltsThe percentage of antiHBV T lymphoeytes with specificity to GLSPTVWLSV in total CD8+ T lymphoeytes from four patients with hepatitis B associated HCC was 1.44％±0.04％,which was higher than those to other four HBV antigen peptides (0.68％±0.08％ of FLLTRILTI,1.06％±0.09％ of FLPSDFFPSV,0.56％ ±0.04％ of FLLSLGIHL,and 0.46％ ±0.08％ of WLSLLVPFV) (t=0.001,P＜0.05).The two lines of monoclonal cell with specificity to GLSPTVWLSV both exhibited high level of IFN-γ expression after incubated with hepatic carcinoma cell line HepG2 (HLA-A2+)with HBV GLSPTVWLSV peptide.ConclusionsCD3+ CD8+ HLA-A2+ cells with specificity to the different HBV peptides exist in PBMC of patients with hepatitis B associated HCC.The expression level depends on HBV antigen peptide sequences and genomic sites.

Objective To investigate the relationship among Helicobacter pylori(H.pylori),CD4 positive cells and CD8 positive cells in gastric mucosa of the AIDS patients with gastritis.Methods Fiftyeight AIDS patients with upper abdominal pain were diagnosed with chronic gastritis through gastroscopy.The gastric biopsies from them were used for H.pylori detection with rapid urease test and Giemsa staining,pathology examination with HE staining,and immunohistochemistry analysis for CD4,CD8 positive cells in Gastric mucosa.And the application of flow cytometry was for the detection of peripheral blood CD4 and CD8 lymphocytes from the patients.Results H.pylori was positive in 26 cases,and negative was in 32 cases.CD8 cell expression in gastric mucosa of the AIDS patients with H.pylori positive was significantly higher than H.pylori negative patients(P＜0.05).There is no difference CD4 cell expression in gastric mucosa between the AIDS patients with H.pylori positive and H.pylori negative patients.Moreover,CD8 positive lymphocytes in gastric mucosa of those patients with H.pyloriinfection were significantly stronger than the CD4 positive lymphocytes.However,the peripheral blood CD4 lymphocytes from the patients with H.pylori infection were more than those from H.pylorinegative patients significantly(P＜0.05).Conclusion The expression level of CD8 cells in gastric mucosal tissues of AIDS patients with H.pylori infection were higher than those without H.pylori infection.The CD4 lymphocytes from the peripheral blood of the patients with H.pylori infection were more than those without H.pylori negative patients.

ObjectiveTo evaluate the accuracy of EUS,EUS-guided fine needle aspiration ( EUSFNA) and targeted biopsy in the diagnosis of wall-thickened gastric lesions with negative malignant results ofendoscopic biopsies.MethodsA retrospective study was carried out in 57 patients who were found with thickened gastric wall of negative malignant endoscopic biopsies and underwent EUS from January 2008 to December 2010 in our hospital.Compared the EUS findings with the surgical results and follow-up status.The diagnostic yield of EUS was characterized by the disappearance of the layers or the changes of thickness of gastric wall,the characteristics of echo imaging,and the results of EUS-FNA or EUS targeted biopsy were recorded to evaluate the value of EUS.ResultsOf 57 cases,gastric cancer was confirmed in 19,lymphoma in 10,dysplasia in 1,Menetrier's disease in 1 and inflammatory changes in 26.EUS could clearly demonstrate the changes of gastric wall including the thickness and the changes of layers,with the accuracy rate of 73.07％ ( 14/19 ) on gastric cancer.EUS diagnosed gastric cancer in 26 cases,in which 14 (53.8％) were confirmed by pathology.Gastric lymphoma was suspected by EUS in 20 cases,in which 10 (50.0％ ) were proved.EUS-FNA was conducted in 19 cases,with positive result in 9 (accuracy rate 50％ ).EUS-guided targeted deep biopsy or piece-meal biopsy were performed in 10 cases,with 8 malignant results.ConclusionEUS with/without EUS-FNA is not a golden standard for the diagnosis of gastric lesions in thickened gastric wall,yet it still has some significance.

ysis, which had important reference values for further studying biological functions of UP Ⅱ gene and targeted therapeutic strategy for TCC of bladder.