Objective To develop quality control methods for the key quality properties of recombinant monoclonal antibodies against proprotein convertase subtilisin/Kexin 9(PCSK9). Methods According to the corresponding requirements of Chinese Pharmacopoeia(Volume Ⅲ,2020 edition)and International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use(ICH),the PCSK9 monoclonal antibody was identified for the identity by reducing trypsin peptide fingerprint mapping analysis,identified for the specificity by ELISA,determined for the charge heterogeneity by imaging capillary isoelectric focusing electrophoresis(icIEF),detected for the variant purity by reducing/non-reducing capillary electrophoresis-sodium dodecyl sulfonate(CE-SDS),measured for the content of monomers,high molecular weight substances and low molecular weight substances by size-exclusion chromatography high performance liquid chromatography(SEC-HPLC),analyzed for the glycotype with N-glycan labeled with 2AB after carbohydrate cutting by Waters ACQUITY UPLC system with fluorescence detector,determined for the biological potency with HepG2 cell line and low-density lipoprotein(LDL),and detected for the content of polysorbate 20 by the liquid phase detection system(CAD detector). Results PCSK9 monoclonal antibody sample had characteristic peptide segments,and the peptide fingerprint was consistent with that of reference substance. The sample contained specific antibodies;The relative area percentages of main peak,acid peak and alkaline peak of the sample were(49. 27 ± 0. 38)%,(46. 44 ± 0. 35)% and(4. 28 ± 0. 04)%,respectively. The relative area percentages of“heavy chain + light chain”peak,non-glycosyl main peak and low molecular weight substance peak were(94. 16 ± 0. 82)%,(4. 11 ± 0. 76)% and(0. 85 ± 0. 20)%,respectively. The relative area percentages of main peak,nonglycosyl main peak and low molecular weight substance peak were(94. 27 ± 0. 22)%,(2. 85 ± 0. 08)% and(2. 88 ± 0. 15)%,respectively. The relative area percentages of monomer,high molecular weight substance and low molecular weight substance were(98. 30 ± 0. 03)%,(0. 75 ± 0. 01)% and(0. 96 ± 0. 02)%,respectively. The relative percentages of G0F,G1F,G2F and non-fucosylated(G0 + G1 + G2 + Man5)were(39. 31 ± 0. 54)%,(34. 69 ± 0. 41)%,(9. 09 ± 0. 14)% and(12. 61 ± 0. 50)%,respectively. The relative biological potency was(101. 64 ± 3. 61)% of the reference. The content of polysorbate 20 was(0. 012 ± 0. 000 3)%. Conclusion According to the key quality attribute of monoclonal antibody against PCSK9,the corresponding key quality control methods have been established,which can ensure the safety,effectiveness and quality controllability of the product.

Objective To explore the relationship between mean platelet volume(MPV),blood fibrinogen(Fg)level and carotid atherosclerosis in patients with cerebral infarction.Methods 50 patients with cerebral infarction were selected as the research objects,and divided into the carotid atherosclerosis group and without carotid atherosclerosis group depending on whether carotid atherosclerosis,each group 25 cases.The MPV and plasma Fg of the two groups were detected and compared.Results The levels of MPV and blood fibrinogen in the carotid atherosclerosis group[(12.99±1.45)fl,(4.59±1.05)g/L]were higher than those without carotid atherosclerosis group[(11.01±1.21)fl,(3.05±0.51)g/L](t=4.212,-0.387,P=0.000,0.001).Conclusion The relationship between MPV,Fg levels and carotid atherosclerosis in patients with cerebral infarction is close.

Carcinogens ; chemistry ; toxicity ; Chromium ; chemistry ; toxicity ; Chromium Compounds ; chemistry ; toxicity ; Humans ; Occupational Exposure ; analysis

Objective: To investigate the changes of vasoconstrictor and vasodilator functions of isolated thoracic aorta in the rats with obstructive jaundice, so as to study the mechanism for the functional changes of the aorta in rats with obstructive jaundice. Methods: Sixty male SD rats were randomly divided into bile duct ligation (BDL) group and sham operation (SO) group. The thoracic aortas were isolated at 3, 7 and 14 d after surgery and the vasoconstrictor responses induced by high potassium, norepinephrine (NE) and phenylephrine (PE) gradient and the vasodilator response to SNP were tested. Another 40 male SD rats were randomly divided into BDL three-day group, BDL seven-day group, BDL 14-day group and sham operation (SO) group. The expression of α1D-AR protein in the thoracic aorta was assessed by Western blotting. Results: The vasoconstrictor responses of thoracic aorta rings in high potassium, NE, PE, and BDL rats were weaker than those in the SO group. The vasodilator response induced by SNP in the BDL group was greater than that in the SO group. The expression of α1D-AR protein in the BDL group was weaker than that in the SO group, and the expression decreased with the prolongation of obstructive time. Conclusion: The systolic and diastolic functions of isolated thoracic aorta rings are changed in cholestatic rats; the weakened systolic function might be a result of decreased expression of α 1D-AR albumen.

We report a case of a 47-year-old man who firstly complained of throat pain for half a month accompanied with fever. Specialized examination showed tonsils' hypertrophy and the laryngoscope found his right vocal cord was swelling and hyperemia. The routine blood test counted white blood cell as 31 x 10(9)/L, lymphocyte as 30. 84 x 10(9)/L while prolymphocyte could be seen with microscope. After that B-ultrasound scan gave spleen hypertrophy and multi-lymphatic-node enlargement. Peripheral blood flowcytometry and bone marrow biopsy finally diagnosed the man as actue myelocytic leukemia.
Humans ; Leukemia, Myeloid, Acute ; complications ; diagnosis ; Male ; Middle Aged ; Pain ; etiology ; Pharynx

Objective To explore the effect of aspirin resistance on cardiovascular events in non-insulin-dependent diabetes melli-tus patients .Methods 496 patients with non-insulin-dependent diabetes mellitus received daily aspirin therapy (100mg/d) 4 weeks were recruited from Jun .2006 to Sept .2010 .Platelet aggregation was measured by light transmission aggregometry .The patients were divided into three groups :aspirin resistance group ,aspirin semi-resistance group and aspirin-sensitive group .The clinical bio-chemistry indications and the adverse cardiovascular events were collected .Results 11 .69% of patients were found to be resistant to aspirin therapy ,and 37 .90% of patients to be aspirin semi-responders .There were 67 .48% female in aspirin resistance group and aspirin semi-responders .The preprandial glucose of group AR and ASR was higher than that of group AS (P<0 .05) .The HbA1C and TG of group AR were higher than those of group AS(P<0 .05) .There was statistically significant difference of the rate of car-diovascular events .Conclusion The measure of platelet aggregation should be taken in the diabetic mellitus patients and pay atten-tion on the dose of aspirin recommended .Particular attention is should also devote to the incidence of adverse cardiovascular events .

BACKGROUND:Although core decompression can contribute to the reconstruction of necrotic bone, its single use appears to have some deficiencies and cannot completely realize the reconstruction of the femoral head.
OBJECTIVE:To explore the effect of arthroscopic core decompression with autologous bone marrow induction material combined with titanium rod in the repair of stage II femoral head necrosis
METHODS:A retrospective analysis was performed on clinical data of 79 cases of stage II femoral head necrosis admitted at the South Branch of Tongchuan People’s Hospital from February 2011 to February 2013. According to the therapeutic methods, these cases were divided into control group (40 cases) and observation group (39 cases), and were given conventional core decompression and arthroscopic core decompression with autologous bone marrow induction material combined with titanium rod. Patients were fol owed up for 24 months, and the range of hip flexion and Harris scores on hip function and adverse events were observed and compared between two groups.
RESULTS AND CONCLUSION:Al the 79 patients successful y completed the 24-month fol ow-up. By the last fol ow-up, the range of hip flexion and Harris score were significantly higher in the observation group than the control group (P<0.05). There was no failure case due to surgical material rejection. To sum up, arthroscopic core decompression with autologous bone marrow induction material combined with titanium rod is safe and effective for repair of stage II femoral head necrosis.

Objective To study the effect of CO 2 pneumoperitoneum and enteric disturbance on serum ?-endorphin (?-EP) in SD rats, in order to investigate their influence on peri-operative stress responses in SD rats. Methods A total of 120 SPF-grade male SD rats were randomly divided into four groups with 30 rats in each group. The four groups received CO 2 pneumoperitoneum (Group A), a 5 cm abdominal incision (Group B), a 5 cm abdominal incision with gastroenteric disturbance (Group C), and intraperitoneal anesthesia (Group D or Control Group), respectively. Concentrations of serum ?-EP of these groups were measured 10 min, 20 min, and 40 min after the beginning of surgery, respectively. Results Concentrations of serum ?-EP in the Group A 10 min, 20 min, and 40 min after the beginning of surgery were 274 7?66 6 pg/dl, 157 3?63 8 pg/dl, and 163 9?74 5 pg/dl, respectively, which were all extremely significantly higher than those in the Control Group ( P

Objective To assess the efficacy and safety of stenting ureteral stent prior to extracorporeal shock wave lithotrip‐sy(ESWL) for non‐obstructive upper ureter and renal stone .Methods From January 2012 to October 2013 ,a total of 96 patients who met inclusion and exclusion criteria were randomly devided into two groups :a stented group (group B) with a double‐J ureter stent fixed pre‐ESWL and a non‐stented group (group A) treated with ESWL direactly .Outcomes were evaluated in 1 month and then the ureter stent was removed ,another evaluation was taken after 2 months .Results The clearance rate in group A was signifi‐cant higher than that of group B at first month(P=0 .02) ,but group B was significant higher than that of group A at the second month(P=0 .001) .The renal colic rate of group A was significant higher than that of group B(P=0 .001) ,the gross hematuria rate of group B was significant higher than that of group A(P=0 .005) .The comparison of fever ,steinstrasse ,lower urinary tract syn‐drome showed no statistical difference(P>0 .05) .Conclusion Stenting ureter stent may increase the clearance rate of ESWL by 1 session for non‐obstructive upper ureter and renal stone ,but the passage of stones mainly happen after the removing of stent .Stent‐ing can also prevent the morbidity of renal colic without increasing low urinary tract syndrome ,but can increase the morbidity of gross hematuira .

Objective:To investigate the effect of Matrine on inhibiting lipopolysaccharide( lipopolysaccharide ,LPS) -induced macrophages to secrete inflammatory cytokines,including tumor necrosis factor-α( TNF-α) ,leukocyte mediated element 1β( IL-1β) on Toll like receptor 4 ( TLR4 ) , c-jun expression. Methods: Cultured macrophage RAW264. 7 of mouse and divided into four groups, including blank control group,matrine group,LPS group,matrine intervention group. Incubated by the concentration was 100 μg/L LPS DMEM for 1 h,then the LPS was discard. Added free serum DMEM or 125. 25 mg/L matrine DMEM to culture. The cells and culture solution of 5,30,60,120 min after the treatment of the above four groups were collected respectively. Detected of mouse RAW264. 7 macrophage TLR4 and c-jun mRNA expression by PCR. Detected of mouse macrophage RAW264. 7 cells c-jun protein expression by immunocytochemical. Measured secretion in cultured solution of TNF-α and IL-1β by ELISA. Results:Expression TLR4 mRNA,c-jun mRNA and c-jun protein and TNF -α and IL-1β secretion quantity indexes of matrine induced and blank group had no statistical significant differences ( P>0. 05 ) . TLR4 mRNA, c-jun mRNA and c-jun protein expression secretion of and TNF-α and IL-1β LPS group at each time point were significantly higher than that of blank group,and the high level to maintain more than 2 h (P<0. 05);matrine intervention group could effectively inhibit LPS induced macrophage TLR4 mRNA,c-jun mRNA and c-jun protein expression and reduce the secretion of inflammatory cytokines TNF-α and IL-1β. Conclusion: Matrine may inhibit LPS induced macrophage mitogen activated protein kinase signal pathway of TLR4 and c-jun expression so that it can effectively reduce the end inflammatory cytokines TNF-α and IL-1β release,effectively inhibit the inflammatory reaction and reduce the degree of endotoxin inflammatory re-sponse.