Objective：This study was designed to express chimeric anti-VEGF (vascular endothelial growth factor) antibody in dihydrofolate reductase-deficient Chinese hamster ovary (CHO-dhfr-)cells at high-level, and explore an optimum method to obtain high-level expression cells clone. Methods：The light chain and heavy chain genes of chimeric anti-VEGF antibody were induced into CHO-dhfr-cells using a novel eukaryotic high-level expression vectors system for genetic engineering antibodies. High-level expression was achieved after subcloning and several rounds of co-amplification of methotrexate (MTX). Biological features and productive amount of chimeric antibody was charactered by ELISA. Result：The cells strain that secret anti-VEGF chimeric antibody at the highest level of 28 μ g/ml was established. The cells were subcloned following each round of co-amplification of MTX, while greatly different results were obtained using three methods. The chimeric antibody contained constant regions of human immunoglobin and had the specificity against VEGF by ELISA. Conclusion：The anti-VEGF mouse-human chimeric antibody was expressed at high-level successfully in CHO cells. This may be an optimum method to obtain high-level expression cells clone for the eukaryotic high-level expression vectors system.

OBJECTIVETo investigate the removal of cadmium from aqueous solution by waste fungal biomass of Aspergillus niger, originated from citric acid fermentation industry.

METHODSBatch adsorption test was used to study the biosorption equilibrium and isotherm. The Cd2+ concentration was measured with atomic adsorption spectrophotometer (AAS) HITACHI 180-80.

RESULTSThe biosorption achieved equilibrium within 30 min. The adsorption isotherm could be described by Freundlich adsorption model, and the constants K(F) and 1/n were determined to be 2.07 and 0.18, respectively, and the correlation efficiency was 0.97. The optimal pH for Cd adsorption was 6.0. The cadmium-laden biomass could be effectively regenerated using 0.1 N HCl.

CONCLUSIONThe waste biomass of Aspergillus niger, a by-product of fermentation industry, is a potential biosorbent for the removal of cadmium from aqueous solution.

Adsorption ; Aspergillus niger ; Biomass ; Cadmium ; isolation & purification ; Citric Acid ; Hydrogen-Ion Concentration ; Industrial Waste ; Waste Disposal, Fluid ; Water Pollutants, Chemical ; isolation & purification ; Water Purification ; methods

OBJECTIVETo evaluate the role of serum level of procalcitonin (PCT), C-reactive protein (CRP), erythrocyte sedimentation rate (ESR), and white blood cell count (WBC) as predictors in postoperative early infectious complications with fever after posterior lumbar internal fixation (PLIF).

METHODSA retrospective study was conducted from January 2012 to January 2014. Fifty-two patients with fever in the early stage(within 10 days) after the PLIF were collected in the study. They were divided into infection group and non-infection group (group A and group B) according to the results of postoperative blood culture. There were 26 patients in group A and 32 patients in group B. The values of PCT, CRP, ESR, and WBC count were compared and analyzed between two groups.

RESULTSThe values of PCT, CRP, and ESR in group A were higher than those of group B. Meanwhile, CRP and ESR in group B were still higher than the normal range. Among the 26 patients with infections (group A), PCT was superior to CRP and ESR, had a good ability in discriminating different kinds of postoperative infections. The area under the ROC curve of serum PCT levels was the largest (CI 95% was 0.81 to 0.98) in the indexs; and ROC curve of WBC count was no statistically significant. When the cut off points of each predictors were evaluated, the higher sensitive was CRP and reached at 90.27% and the higher specific was ESR and reached at 88.50%.

CONCLUSIONFor the patients with fever at the early stage after the PLIF should be paid attention and reasonable choosing predictors are helpful to identify postoperative infection in the early stage. The CRP and ESR may be influenced by the surgery, and the PCT level is helpful to differentiate infection type.

Adult ; Aged ; Aged, 80 and over ; Blood Sedimentation ; C-Reactive Protein ; analysis ; Calcitonin ; blood ; Calcitonin Gene-Related Peptide ; Female ; Fever ; blood ; diagnosis ; Fracture Fixation, Internal ; adverse effects ; Humans ; Infection ; blood ; diagnosis ; Leukocyte Count ; Lumbar Vertebrae ; surgery ; Male ; Middle Aged ; Postoperative Complications ; blood ; diagnosis ; Protein Precursors ; blood

OBJECTIVETo explore therapeutic effect and action mechanism of regulating spleen-stomach needling on diabetic nephropathy (DN).

METHODSUsing multi-centric, randomized, controlled and blind principles, 144 cases of DN were divided into an observation group and a control group according to random digital tab, 72 cases in each one. Based on regular treatment of diabetes, the regulating spleen-stomach needling was applied at Zhongwan (CV 12), Quchi (LI 11), Hegu (LI 4) and Xuehai (SP 10), etc. in the observation group while Shenshu (BL 23), Taixi (KI 3), Sanyinjiao (SP 6), Yanglingquan (GB 34), etc. were selected in the control group by reference of Acupuncture and moxibustion. The treatment was given twice a day, six days as a treatment session with interval of one day between sessions. Totally six weeks were required. Changes of clinical symptoms and signs, fast blood glucose (FBG), urinary albumin excretion rate (UAER), beta2-microglobulin (beta2-MG), monocyte chemotactic protein-1 (MCP-1), lymphocyte membrane cholesterol, propanediol (MDA), PCO, 8-hydroxydeoxy guanosine (8-OHdG), superoxide dismutase (SOD), CD3+, CD4+, CD8+, and CD4+/CD8+ were observed before and after treatment in two groups.

RESULTSAs for improving clinical symptoms and signs, total effective rate was 84.29% (59/70) in the observation group and 55.56% (40/72) in the control group, which had statistical difference between two groups (P<0.01). As for regulating glycometabolism [(6.25 +/- 0.32) mmol/L vs (8.09 +/- 0.63) mmol/L], reducing UAER [(154.43 +/- 55.14) mg/24h vs (268.91 +/- 77.65) mg/24h], restraining over-expression of MCP-1 [(137.59 +/- 36.15) pg/mL vs (166.89 +/- 42.82) pg/mL], regulating level of oxidative stress, prohibiting oxidation of protein and adjusting quantity and activity of T lymphocyte subgroup, the observation group was superior to the control group (P< 0.05, P<0.01).

CONCLUSIONThe regulating spleen-stomach needling is an effective method for treatment of DN, which cold improve glycometabolism disturbance-induced progressive kidney injury, recover glomerular filtration, reduce urinary albumin excretion rate, restrain overexpression of MCP-1, adjust level of oxidative stress, prohibit oxidation of protein, increase protectiveness of membrane, adjust quantity and activity abnormity of T lymphocyte subgroup, leading to repairing lymphocyte damage and improving immune expression to delay kidney damage.

Acupuncture Points ; Acupuncture Therapy ; Adult ; Aged ; Diabetic Nephropathies ; immunology ; physiopathology ; therapy ; Female ; Humans ; Male ; Middle Aged ; Spleen ; physiopathology ; Stomach ; physiopathology

OBJECTIVETo evaluate clinical outcomes of anterior cruciate ligament (ACL) and posterior cruciate ligament (PCL) reconstruction under arthroscopy combined with limited open repair of medial collateral ligament (MCL) for the treatment of multiple ligament injuries of knee joints.

METHODSFrom March 2006 and June 2012,the data of 14 patients (14 knees) with multiple injuries of ACL, PCL, and MCL were collected. There were 8 males and 6 females with an average age of (31.8 +/- 8.1) years old (ranged, 20 to 49 years old). All the patients were performed with X-ray and MRI examination, and the results showed that 10 patients had combined with injuries of anterior cruciate ligament (ACL), posterior cruciate ligament (PCL) and medial collateral ligament (MCL); 4 patients had ALC,PCL and posterolateral corner (PLC) injuries. Four patients had medial meniscus injuries and 2 patients had lateral meniscus injuries. The MCL,PLC and meniscus injuries were treated with operation on the first stage, and functional exercises were performed 3 weeks after fixation. The reconstruction operation of ACL and (or) PCL was performed at the second stage under arthroscopy 3 to 6 months later when the movement range of knee joint recovered to the normal level with obvious relaxation.

RESULTSAll incisions healed by primary intention. All the patients were followed up with a mean duration of 48.9 months (ranged, 24 to 80 months). The Lysholm score was improved from preoperative 19.6 +/- 0.9 to the latest follow-up 87.1 +/- 2.8 (t=12.3, P<0.01). The International Knee Documentation Committee (IKDC) rating: 9 cases nearly recovered to normal, 5 cases were abnormal.

CONCLUSIONFor multiple ligament injuries in the knee, staged repair and reconstruction can effectively restore knee joint stability and function.

Adult ; Anterior Cruciate Ligament ; surgery ; Anterior Cruciate Ligament Injuries ; Female ; Follow-Up Studies ; Humans ; Knee Injuries ; physiopathology ; surgery ; Knee Joint ; physiopathology ; surgery ; Male ; Middle Aged ; Posterior Cruciate Ligament ; injuries ; surgery ; Reconstructive Surgical Procedures ; Treatment Outcome ; Young Adult

Objective To investigate the relationship of Vaneomycin serum concentration and drug adverse reactions (ADRs) in neonates.Methods Two hundred and thirteen neonates who were treated by Vancomycin and had their serum concentration monitored were recruited.The number of cases which had liver,kidney and hearing damage after treatment was calculated.The correlation between 3 different serum concentrations (＜5 mg/L,5-10 mg/L,＞ 10 mg/L) and ADRs were analyzed.Results The mean level of Vancomycin in serum was (4.36 ± 4.99) mg/L The total incidence of ADRs was 17.8％ (38/213 cases).The main ADRs were liver damage and hearing impairment,whose incidences were 9.9％ (21/213 cases) and 7.5％ (16/213 cases),respectively,and they were were significantly higher than kidney danage (0.5％,1/213 cases) (x2 =19.172,P =0.000;x2 =13.785,P =0.000).There was no significant difference be-tween the inidence ratio of liver damage and that of the hearing impairment (x2 =0.330,P =0.566).The ADRs ratio among 3 different serum concentrations was 17.0％ (24/141 cases),17.6％ (9/51 cases) and 23.8％ (5/21 cases).There was no significant difference between the ADRs ratios and distribution of different serum concentrations (x2 =0.576,P =0.750).Moreover,the ADRs rates didn't increase along with the serum concentrations (Z =0.648,P =0.517).Except for aspartate transaminase,the indicators of liver and kidney function varied significantly after vancomycin treatment (P ＜0.05).But there was no clinical significance because the mean value was in normal range.By Bivariate Correlation analysis,Vancomyein serum concentration had no significant influence on liver and kidney function (P ＞ 0.05).Conclusions Neonates treated by Vancomycin had reatively high ratio of the liver damage and the hearing impairment.There is no significant correlation between Vancomyein serum concentration and ADRs in neonates.

Objective To study CT-guided localization of additional pulmonary nodules with microcoils prior to video-assisted thoracoscopic surgery (VATS) resection in patients with suspected lung cancer.Methods Eleven patients suspected lung cancer underwent preoperative microcoils localization towards additional small pulmonary nodules.The head of microcoil was pinpointed adjacent to the target nodule while its end tail remained above the visceral pleura.VATS were performed within 24 hours, and comprehensive assessments were conducted according to surgical and pathological outcomes of primary and additional lesions, and suitable surgical processes were followed.Results All 11 localizing pulmonary nodules (4-15 mm in diameter) were successfully removed after VATS, 9 microcoils'' end tails of which were placed above visceral pleural surface.There were no serious complications related with localizing procedure.Other 16 lesions including 11 primary ones were resected.The surgical and pathological outcomes for lung lesions were utterly assessed.Conclusion Microcoil preoperative localization provides helpful orientation for complete resection and assessment of multiple pulmonary lesions in patients with suspected lung cancer.

Objective: To observe the changes in numbers and degranulation status of local mast cells (MCs) of the complementary acupionts, as well as the concentrations of serum histamine and bradykinin after acupuncture at rat's complementary acupionts of Yanglingquan (GB 34) and Yinlingquan (SP 10), for exploring the mechanisms of the synergistic effect of complementary acupionts. Methods: Using random number table method, 40 Wistar rats were randomly divided into a blank control group (group K), an acupuncture at Yanglingquan (GB 34) group (group A), an acupuncture at Yinlingquan (SP 9) group (group B), an acupuncture at Yanglingquan (GB 34) and Yinlingquan (SP 9) group (group AB), and an acupuncture at points near Yanglingquan (GB 34) and Yinlingquan (SP 9) group (group CD), 8 rats in each group. Group A received acupuncture at Yanglingquan (GB 34); group B received acupuncture at Yinlingquan (SP 9); group AB received acupuncture at Yanglingquan (GB 34) and Yinlingquan (SP 9); group CD received acupuncture at the control points [points 3 mm away from Yanglingquan (GB 34) and Yinlingquan (SP 9) respectively]. Acupuncture was performed at bilateral points in each group. Rats in group K were fixed using the same method as rats in the other 4 groups without acupuncture stimulation. Needles of 0.35 mm in diameter and 40 mm in length were used for acupuncture. Needle handle was connected to the G6805-Ⅱ mode electroacupuncture (EA) device after needling qi was obtained, with sparse-dense wave, frequency of 2 Hz/100 Hz and current of 1 mA to keep the needle handles slightly tremulous while the rats kept quiet. Rats were continuously stimulated by EA for 20 min each time. Experimental interventions were conducted on the 1st, 3rd, 5th and 7th days when the experiment started, for 4 times in total. Specimens from rats in each group were collected 2 h after the 7th day. The levels of histamine and bradykinin in serum were detected by enzyme-linked immunosorbent assay (ELISA). Local tissues of the points were used to prepare cryosections. The changes of MCs were observed after toluidine blue staining. Results: Compared with group K, the numbers and degranulation rates of MCs in group A, group B, group AB and group CD were significantly increased (all P<0.05). The number and degranulation rate of MCs in group AB were significantly higher than those in group A, group B and group CD (all P<0.05). The order of histamine levels from high to low was: group AB >group B > group A > group CD > group K, and the differences were statistically significant (all P<0.05); the levels of bradykinin in group AB, group A and group B were significantly higher than those in group K and group CD (all P<0.05). There was no significant difference in the bradykinin level between group A and group B, nor between group CD and group K (all P>0.05). Conclusion: The number and degranulation rate of MCs of the complementary acupionts are significantly increased after acupuncture at complementary acupionts of Yanglingquan (GB 34) and Yinlingquan (SP 9) of rats, and the serum levels of histamine and bradykinin are increased, which may be one of the mechanisms of the synergism produced by the combination of complementary acupionts.

Coronary Sinus ; abnormalities ; Humans ; Male ; Middle Aged ; Subclavian Vein ; Vena Cava, Superior

OBJECTIVETo study the toxicity on skin and penetration effect of volatile oil from tender branchers of Camellia oleifera on nitrendipine, baicalin, nimesulide for percutaneous obsorption.

METHODAcute skin toxicity, irritation and allergy on rats were tested, and mouse skin in vitro was applied for studying the effects of different concentrations of volatile oil in nitrendipine, baicalin, nimesulide on drug permeation.

RESULTDifferent dosage volatile oil had no acute toxicity, irritation or hypersensitive effects. Compared to azone, more powerful enhancement effects of volatile oil at different concentration on nitrendipine, baicalin, nimesulide were very obvious.

CONCLUSIONThis paper firstly reported the results of experiment about the toxicity to skin and penetr-ation effect of volatile oil from tender branches of C. oleifera.

Administration, Cutaneous ; Animals ; Camellia ; chemistry ; Female ; Flavonoids ; administration & dosage ; pharmacokinetics ; In Vitro Techniques ; Male ; Mice ; Nitrendipine ; administration & dosage ; pharmacokinetics ; Oils, Volatile ; isolation & purification ; pharmacology ; toxicity ; Permeability ; drug effects ; Plant Oils ; isolation & purification ; pharmacology ; toxicity ; Plant Stems ; chemistry ; Plants, Medicinal ; chemistry ; Rats ; Rats, Sprague-Dawley ; Skin ; drug effects ; metabolism ; Skin Absorption ; drug effects ; Sulfonamides ; administration & dosage ; pharmacokinetics