Objective To assess the clinical effect and safety evaluation of Keluoxin capsule in the treatment of the patients with early diabetic nephropathy. Methods One hundred and forty-nine patients with early diabetic nephropathy were randomly divided into the treatment group and control group, 74 in each group. The control group was treated with Keluoxin placebo capsules (2 g every time, 2 times a day, oral) and Irbesartan (150 mg every time, once a day, oral) simultaneously; and thetreatment group was treated with Keluoxin capsules (2 g every time, 2 times a day, oral) and Irbesartan (150 mg every time, once a day, oral) simultaneously. Both groups were treated for 24 weeks. The clinical curative effect, the changes of blood sugar levels, TG, TC, LDL-C, HDL-C, BUN, Scr, 24 hUTP and 24 hUAE were assessed and compared, and the treatment safety of Keluoxin capsule was assessed. Results Compared with the control group, the markedly effective rate and total effective rate of treatment group increased significantly (χ2=9.208, P<0.05). The levels of blood sugar (7.4 ± 0.6 mmol/L vs. 6.2 ± 0.5 mmol/L, P<0.05), TC (7.1 ± 2.3 mmol/L vs. 5.4 ± 1.6 mmol/L, P<0.05), TG (2.7 ± 0.8 mmol/L vs. 2.3 ± 0.5 mmol/L, P<0.05), LDL-C (4.2 ± 1.6 mmol/L vs. 3.5 ± 1.2 mmol/L, P<0.05), BUN (11.4 ± 3.3 mmol/L vs. 6.4 ± 2.5 mmol/L, P<0.05), SCr (158.7 ± 29.5μmol/L vs. 116.2 ± 23.1μmol/L, P<0.05), 24 hUTP (237.1 ± 46.3 mg vs. 201.4 ± 38.2 mg, P<0.05) and 24 hUAE (157.3 ± 40.8 mg vs. 109.4 ± 28.5 mg , P<0.05) in the treatment group were significantly lower than those in the control group;but the level of HDL-C showed no significantly difference between two groups (1.1 ± 0.4 mmol/L vs. 1.3 ± 0.6 mmol/L, P>0.05). There was no adverse event between two groups. Conclusions The clinical effect of Keluoxin capsule can benefit the patients with early diabetic nephropathy.

Objective To establish a gas chromatography ( GC) method for determination of aniline,2-aminophenol and 4-aminophenol in mesalamine sustained release granules. Methods An HP-5 (10 m×0.53 mm,2.65 μm) capillary GC column was adopted. The carrier gas was high purity nitrogen at a flow rate of 15 mL.min-1 . The inlet temperature was controlled at 280 ℃ and the FID detector temperature was 300℃. The oven temperature was initially held at 70℃ for 2 min and was then programmed to 150 ℃ at 10℃.min-1 and held for 6 min. Results A good resolution was obtained between the peaks of the 3 impurities at chromatographic conditions above. The recoveries of aniline, 2-aminophenol and 4-aminophenol were among 98.04% to 103.00%. The precision and linearity equations were good. Conclusion The method can be adopted for the quality control of mesalazine sustained release granules.

Objective To explore the effect of berberine hydrochloride on serum HIF-1α,Caspase -3 levels in patients with acute cerebral infarction with a multicenter study.Methods 126 patients with acute cerebral infarction from January 2012 to January 2014 were selected in the Affiliated Hospital of Hebei Engineering University,the First Hospital of Handan City,Handan Central Hospital were divided into experimental groups and control group using random number table.Control group had 63 cases,were given 80 mg ozagrel sodium injection intravenously,twice a day,and 800 mg calf blood protein injection intravenously,once per day on the basis of conventional treatment;at the same time,100 mg of oral aspirin tablets were taken,once per night,7 days for a course of treatment.Experimental group were given berberine hydrochloride on the basis of conventional treatment,0.7 g orally,three times a day,7 days for a course of treatment.After a course of treatment,serum HIF-1α,Caspase-3 levels in patients with acute cerebral infarction were observed before and after treatment.Results Serum HIF-1αlevels in two groups were reduced after treatment,and its level in experimental group[(733.49 ±225.01)ng/mL]was lower than that in control group [(975.32 ±311.35)ng/mL],the difference was statistically significant (P<0.05 ).Serum Caspase-3 levels in two groups after treatment were decreased,and its level in experimental group [(1 1.88 ±2.66 )ng/mL]was lower than the control group [(12.51 ±3.77)ng/mL],the difference was statistically significant (P <0.05).Conclusion Berberine hydrochloride could improve the serum HIF-1α,Caspase-3 levels in patients with acute cerebral infarction,reduce the fatality rate,and could be actively used in the recovery of patients with acute cerebral infarction treatment.

Objective To evaluate the effect of the difficult embryo transfer on the clinical pregnancy outcome of in vitro fertili-zation-embryo transfer(IVF-ET) .Methods There were 209 fresh cycles of difficultly transferring and 2 489 fresh cycles of easily embryo transferring between January 2011 and December 2012 .The clinical outcome was compared .Results There were statistical-ly significant differences in the catheter blood staining rates (51 .20% vs 27 .68% ,P< 0 .05) ,implantation rate(31 .14% vs 35 . 54% ,P<0 .05) ,and clinical pregnancy rate (46 .41% vs 55 .56% ,P<0 .05)between the two groups .There was no significant difference in the rates of ectopic pregnancy and miscarriage between the two groups (P>0 .05) .Conclusion Difficulty ET will in-fluence the clinical pregnancy .Therefore ,all efforts should be made to avoid the difficult transfer in order to increase the pregnant rate .

Objective To summarize the clinical characteristics,diagnosis and treatment of autoimmune pancreatitis(AIP).Methods The retrospective descriptive study was adopted.The clinical data of 43 patients with AIP who were admitted to the First Affiliated Hospital of China Medical University between January 2010 and December 2014 were collected.All patients underwent laboratory tests and imaging examinations.For the patients who had the definite diagnosis of AIP,corticosteroid therapy was adopted,and for the patients who had the suspicion of pancreatic cancer,surgical operation was carried out.Observed indexes included clinical manifestations,laboratory findings,imaging findings,treatment,pathological findings and follow-up status.The follow-up of imaging examinations and laboratory tests via outpatient and inpatient examinations was performed on the patients until January 2016.Measurement data with skewed distribution were presented as average (range).Results (1) Clinical manifestations:of the 43 patients,33 demonstrated obstructive jaundice,19 had loss of weight,18 had upper abdominal pain and 5 had acute mild pancreatitis (the same patient with a variety of symptoms).Extrapancreatic organ involvement was detected in 23 of the 43 patients,including focal nephritis in 15 patients,sicca syndrome in 7 patients,pulmonary involvement in 5 patients,autoimmune thyroiditis in 5patients,hilar IgG4-associated cholangitis in 3 patients,retroperitoneal fibrosis in 3 patients and ulcerative colitis in 1 patient (the same patient with a variety of diseases).(2) Results of laboratory tests:17 patients underwent serum IgG4 detection and increased expression of IgG4 was found in 15 patients.(3) Imaging findings:all the 43patients underwent enhanced computed tomography (CT) scan of pancreas.Diffused enlargement of pancreas with appearance of sausage was observed in 34 patients and localized pancreatic head enlargement was observed in 9patients.Twenty patients had peripancreatic vascular involvement including stenosis or occlusion of splenic vein,stricture at the confluence of superior mesenteric vein,portal vein and splenic vein resulting in regional portal hypertension which led to gastric venous varices and splenomegaly.Three patients showed that superior mesenteric artery was surrounded by tissue masses with massive ascites.Five patients had pancreatic pseudocyst with splenic vein involvement and peripancreatic effusion.The abdominal CT examination of 15 patients with renal involvement showed renal cortical wedge or nodular lesions.Pulmonary inflammatory pseudotumor and interstitial pneumonia appeared in 5 patients,CT of whom showed solid pulmonary nodules or diffuse ground-glass opacities and patchy shadows in pulmonary interstitium.Thirty-two patients underwent magnetic resonance cholangiopancreatography and 28 patients underwent endoscopic retrograde cholangiopancreatography.The results showed the focal and diffuse or segmental stenosis of the main pancreatic duct were detected in 28 patients (diffuse stenosis in 23patients,local stenosis in 5 patients),stenosis of the intrapancreatic common bile duct in 32 patients,stenosis of hilar bile duct in 3 patients.Nine patients underwent endoscopic ultrasonography examination.The results showed 3 patients had hypoechoic mass in the head of pancreas and 6 patients had diffuse or heterogeneous hypoechoic of the pancreatic parenchyma.(4) Treatment status:of 43 patients,30 patients received initial steroid treatment.After steroid treatment,the clinical symptoms of 27 patients gradually improved and laboratory and imaging findings were both improved.Eleven patients didn 't receive initial steroid treatment,of whom 2 patients with obstructive jaundice underwent endoscopic biliary stent implantation,7 patients underwent surgical biliary drainage and 2 received neither endoscopic treatment nor surgical treatment.Two patients underwent pancreatoduodenectomy due to suspicion of pancreatic adenocarcinoma and were diagnosed as AIP based on postoperative pathological results and treated with steroid after operation.(5) Pathological examination:of the 43 patients,22 underwent pathological examination.Results of pathological examination of superficial lymph nodes showed lymph follicles hyperplasia,and immunohistochemistry staining showed massive IgG4 positive plasmocytes in 5 patients,supporting the diagnosis.Results of fine needle aspiration cytology in 15 patients showed no cytological evidence were detected in 14 patients and epithelial cells of malignant tumors in 1 patient.Results of pathological examination of surgical specimens showed dense infiltration of lymphocyte and immunohistochemistry staining showed massive IgG4 positive plasmocytes.(6) Follow-up status:of 43 patients,41 were followed up for an average time of 28 months (range,12-71 months).Of the 30 patients who received steroid treatment,27 had complete response and 3 had partial response.Of the 27 with complete response,4 patients relapsed during followup and were cured by continuous steroid treatment and extending treatment course,3 patients did not relapse,1 patient relapsed again,and long-term low dose steroid maintenance therapy was needed.The 3 patients with partial response were treated with mycophenolate mofetil for 6,4,4 months and imaging abmormalities disappeaved absolutely.Eleven patients received no steroid for the initial treatment,in whom 2 patients were lost to follow-up,1 patient with common bile duct jejunum anastomosis died in 1 year after surgery.In the other 8 patients,the imaging abnormalities were spontaneous remission or clinical and imaging manifestations were improved after steroid treatment and no recurrence was found.Two patients who underwent pancreatoduodenectomy due to suspicion of pancreatic adenocarcinoma received steroid treatment after operation and no recurrence was observed.Conclusion Clinicians should combine clinical manifestations,serological examination,imaging findings and pathological examination to make definite diagnosis of AIP and make differential diagnosis of pancreatic cancer to avoid unnecessary surgical treatment by misdiagnosis.

Objective To investigate the relationship between serum HBV -DNA quantitative detection level and liver function markers and hepatitis B markers in patients with chronic hepatitis B virus. Methods The clinical data of 80 patients with chronic hepatitis B virus infection were retrospectively analyzed,and the HBV-DNA load,the serum hepatitis B markers and liver function indicators of the patients were detected and recorded. Another 80 cases received normal physical examination were selected as the control group. The serum markers were detected in both two groups. Results Of 80 patients with chronic hepatitis B virus infection,the positive rate of HBV-DNA was 45. 0% (36/80) and the detection rate of HBeAg was 30. 0% (24/80). The positive rate of HBV -DNA was positively correlated with HBeAg positive rate(χ2 =12. 46,P=0. 00). The levels of alanine aminotransferase (ALT),α-L-fucosidase (AFU),aspartate aminotransferase (AST) and plasma prothrombin time (PT) in HBV-DNA positive patients were significantly higher than those in HBV-DNA negative group( F=42. 693,18. 752,32. 692, 48. 225,all P=0. 000);and with the increase of the amount of virus,the level of the patients' index continued to increase. Conclusion For patients with chronic hepatitis B,the HBV-DNA level and the viral replication should be detected,and the liver function should be evaluated to observe the liver damage timely,thus to provide reference for clinical effective intervention.

Objective:To prepare a recombinant hemagglutinin trimer (HA-Tri) vaccine against influenza viruses and to study its immunogenicity in a mouse model.Methods:A stable CHO cell line that could express HA-Tri was constructed. Western blot, single radial immunodiffusion, protein particle size detection and N-glycosylation site analysis were performed for qualitative and quantitative analysis of the recombinant protein. According to the different treatment conditions such as dosage and adjuvant, BALB/c mice were divided into 11 groups and subjected to consistent immunization procedures. Serum neutralizing antibody titers were measured on 56 d after the first immunization to evaluate the immunogenicity of HA-Tri.Results:The constructed CHO cells could secret and express HA-Tri proteins. The HA-Tri proteins were biologically active and capable of forming precipitation rings in the single radial immunodiffusion. The particle size of HA-Tri was approximately 18.79 nm and 10 N-glycosylation sites were detected, including high mannose, complex glycoforms and heterozygous glycoforms. After prime-boost immunization, there was no statistically significant difference in the titers of neutralizing antibodies induced in mice by 3.75 μg of HA-Tri in combination with RFH01 adjuvant and 15 μg of monovalent vaccine stock solution ( P=0.431 2, U=36). Serum antibody titers in the HA-Tri+ RFH01 groups were higher than those in the corresponding HA-Tri groups without RFH01 adjuvant, and the highest titer was induced in the 15 μg HA-Tri+ RFH01 group, which was 1 280. Conclusions:The recombinant HA-Tri protein was successfully prepared. HA-Tri in combination with RFH01 adjuvant could induce humoral immune responses against influenza viruses in BALB/c mice, which would provide reference for the development of influenza virus recombinant subunit vaccines.

Objective:To evaluate the immunogenicity of a quadrivalent subunit vaccine combined with RFH01 adjuvant in a mouse model.Methods:Identification tests were performed on four monovalent influenza virus subunit vaccine stock solutions according to the methods described in Part 3 of the Chinese Pharmacopoeia 2020 Edition. In the study of the quadrivalent subunit vaccine combined with RFH01 adjuvant, 460 female BALB/c mice (6-8 weeks old) were randomly divided into 46 groups including experimental groups, vaccine control group, negative control group and blank group with 10 mice in each group. In the study of the quadrivalent subunit vaccine in old and young mice, 80 female 10-month-old and 80 female 10-week-old BALB/c mice were randomly divided into 16 groups ( n=10) including monovalent influenza virus vaccine group, quadrivalent subunit vaccine group, quadrivalent subunit vaccine+ RFH01 adjuvant group, chicken embryo quadrivalent split vaccine control group and PBS group. All mice were immunized by intramuscular injection. At 21 d after the primary immunization, a booster immunization was conducted using the same strategy. Blood samples were collected at 21 d and 42 d after the primary immunization for serum separation. Haemagglutination inhibition (HI) test was performed to detect the antibody levels in mouse serum samples. Results:After the booster immunization, the positive conversion rates in all vaccine+ RFH01 adjuvant groups reached 100%, and the geometric mean titers (GMTs) of serum antibodies were significantly higher than those of the vaccine groups without RFH01 adjuvant. There were significant differences in serum antibody titers between the monovalent/quadrivalent subunit vaccine groups with and without RFH01 adjuvant. After the booster immunization, the titers of serum antibodies against H1N1, H3N2, B/Victoria and B/Yamagata in the 10-week-old mice were significantly higher than those in the 10-month-old mice.Conclusions:The monovalent and quadrivalent influenza virus vaccines in combination with RFH01 adjuvant could elicit higher antibody titers in young (6-10 weeks old) and old (10 months old) mice, showing good immunogenicity.

Prostate cancer is one of the malignant tumors with high incidence. Although most patients with prostate cancer respond well to standard treatment, they often have a poor prognosis once they develop hormone resistance. Radionuclide targeted therapy is an important method to treat malignant tumors after surgery and chemo-radiotherapy. New radioligand therapy (RLT), represented by 177Lu-prostate specific membrane antigen (PSMA)-RLT, effectively solves the problem of poor efficacy in advanced hormone-resistant prostate cancer, and has been widely recognized in the world. In this paper, the clinical practice of 177Lu-PSMA-RLT in the treatment of prostate cancer and its common adverse reactions are described in order to better understand and master its methods and lay the foundation for better clinical application and follow-up research.

Objective:To purify H5N1 influenza virus concentrate prepared by MDCK cells with a new mixed-mode chromatography medium Capto Core700 and the traditional medium Sepharose 4FF, and to compare the separation and purification efficacy of the two media.Methods:Capto Core700 and Sepharose 4FF were used to purify inactivated H5N1 influenza virus concentrate. The morphology of virus particles in different samples was then observed under a transmission electron microscope. Single radial immunodiffusion (SRID), Folin-Phenol (Lowry) method, double-antibody sandwich ELISA and qPCR were used to detect hemagglutinin, total protein, host cell protein (HCP) and host cell DNA (HCD) before and after purification. The recovery rate of virus antigen and the removal rate of impurities were calculated. The immunogenicity of the viruses purified with different media was analyzed using animal experiments. Difference in the purification efficacy of the two chromatography media was analyzed by t-test. Results:H5N1 influenza viruses purified by Capto Core700 or Sepharose 4FF showed the typical influenza virus morphology under transmission electron microscope. There was no significant difference in the recovery rate of hemagglutinin between the two chromatography media ( P>0.05), but compared with Sepharose 4FF, Capto Core700 had a higher removal rate of impurities (total protein, HCP, HCD) and the difference was statistically significant ( P<0.05). Animal experiments showed that the viruses purified by the two chromatography media had good immunogenicity. Conclusions:Compared with Sepharose 4FF chromatography medium, Capto Core700 could more effectively remove process-related impurities such as HCP, HCD and total protein without affecting the recovery rate of viral antigen. This study provided reference for the development of purification technology in the production of H5N1 influenza virus vaccine in MDCK cells.