An enzyme linked immunosorbent assay ( ELISA ) and a flow cytometry assay ( FCA ) based on Wil2-S cells were developed and systematically compared for quantification of recombinant anti-CD20 humanized monoclonal antibody ( rh-anti-CD20zumab) in biological matrix. The specificity, precision and accuracy of each method at correspondingly different linear range showed good results. For ELISA, the precisions of intra-day and inter-day were both <19 . 5%, the relative error was from-18 . 2% to 17 . 6%;For FCA, the precisions of intra-day and inter-day were both <19. 0%, the relative error was from -18. 9% to 18. 4%. The sensitivity of ELISA was significantly higher than that of FCA. The quantitative ranges of ELISA and FCA methods were 0. 04-5. 0 mg/L and 3. 1-200 mg/L, respectively. The concentrations in serum samples and pharmacokinetics analysis were determined by both of two methods after vein drip administration of rh-anti-CD20zumab in rhesus monkeys. Pharmacokinetics data showed that there was excellent consistency between results obtained by two methods at the given dose. We believe that the novel FCA with high speed and high sensitivity can be used to perform PK and PD study of cell surface antigen-targeted antibody derivatives.

OBJECTIVE To study the pharmacokinetics of heat shock protein 65-mucin 1 (HSP65-MUC1) recombinant fusion protein vaccine in Macaca mulatta monkeys and tumor-bearing mice. METHODS HSP65-MUC1 was labeled by radioactive isotope 125I. M. mulatta monkeys were randomly divided into sc and iv administration groups. Simultaneously, sc administration group was designed as a multiple dose group in which M. mulatta monkeys were sc given [ 125I] HSP65-MUC1 40 μg·g-1, once every 2 weeks for a total of 3 times. Size exclusion chromatography ( SEC) was used to determine concentrations of HSP65-MUC1 in serum samples. The tumor-bearing mice were randomly divided into 0.5, 1.5, 4, 8 and 24 h groups. Mice were sc given [125I] HSP65-MUC1 550 μg·kg-1, tissues were collected and tissue distribution of [125I] HSP65-MUC1 in tumor-bearing mice was studied using trichloroacetic acid (TCA) precipitation method. RESULTS The absolute bioavailability of [125I]HSP65-MUC1 was 38.33% after M. mulatta monkeys were sc given [125I]HSP65-MUC1. In multiple dose group, concentrations of [125I]HSP65-MUC1 after the third dose administration was compared to that of the first dose administration. The accumulation factor (AUC3/AUC1) was 1.17 ±0.25. Distribution of [ 125I]HSP65-MUC1 was significantly different compared with general polypeptide and protein drugs after sc in tumor-bearing mice. The concentration in lymph nodes was the highest. The concentration in other immune tissues, such as thymus and spleen, were not relatively high, but their declined tendency was slow after reaching the peak concentration (cmax ). However, the concentrations in the serum and some other tissues with a large blood volume, such as the heart, liver, and lung, were relatively low and declined quickly after reaching cmax. Its level in the tumor was not very high. [125 I] HSP65-MUC1 was excreted mainly by the kidneys. CONCLUSION The bioavailability of [125I]HSP65-MUC1 is 38.33% after sc administration in M. mulatta. After multiple-dose administration, the vaccine does not accumulate in the body, whose concentration is the highest in lymph nodes after [1251] HSP65-MUC1 was sc given in tumor-bearing mice, but is not very high in tumor. Besides, the vaccine declined tendency is slow after reaching cmax in immune tissues such as thymus and spleen compared with other tissues with a large blood volume.

OBJECTIVE To investigate the feasibility and application of enzyme-linked bridging assay(ELBA)method to the pharmacokinetic evaluation of antisense strand siRNA drug. METHODS Antisense strand RNAs were diluted in LNCap cell lysates from 5 to 50 000 pmol·L-1 to construct the quantification curves. We transfected the intact double-strand siRNA at a final concentration 100 nmol·L-1 targeting Polo-like kinase into the LNCap cells and investigated the specificity of ELBA quantitating the siRNA antisense strand in cell supernatant,cell lysates and RNA-induced silencing complex( RlSC). Quantification curves were constructed and validated in biological matrices such as plasma (5-25 000 pmol·L-1 )and multiple tissues(liver,heart,spleen,and kidneys)(3-6250 pmol·L-1 ). The prostate specific membrane antigen aptamer siRNA delivery system with the intact siRNA concentration of 15 nmol·kg-1 was prepared. The siRNAs were delivered into the LNCap xenogrant tumor model in C57 mice by tail vein injection. The concentration of siRNA antisense strand was determined in plasma and tissues 30 min post administration by ELBA. RESULTS The quantitative range of antisense strand siRNA in cell lysates was 5-50 000 pmol·L-1 ,and ELBA method could quantify the siRNA antisense strand concentration from cell lysates and RlSC in LNCap cells transfected with double-strand siRNA. ln addition,ELBA could specifically reflect the single antisense strand concentration instead of intact siRNA double strands in plasma. The quantification range of siRNA antisense strand using ELBA in plasma was 5-25 000 pmol·L-1 and 3-3125 pmol·L-1 in tissues. About 30 min post administration of PSMA aptamer-siRNA,the antisense strand of siRNA was distributed mainly to the tumor,liver,kidneys,blood and spleen in sequence. The distribution profile might be attributed to the target delivery and siRNA pharma-codynamics. CONCLUSION The ELBA method is successfully applied to the siRNA antisense strand pharmacokinetic evaluation,which provides an alternative for pharmacokinetic studies of siRNA-based drugs.

Objective To identify the risk factors for early postoperative cognitive dysfunction (POCD) in the elderly patients undergoing spinal surgery.Methods One hundred and fifty ASA Ⅱ or Ⅲ patients,aged ≥65 yr,undergoing elective spinal surgery under general anesthesia,were studied.Venous blood samples were taken at 1 day before operation and 7 days after operation to determine the concentration of serum S-100B protein.Cognitive function was assessed by Mini-Mental State Examination (MMSE) at 1 day before operation and 7 days after operation.The patients were diagnosed as having POCD if MMSEpre-MMSEpost ≥ 3.The patients were divided into POCD group and non-POCD group.Age,body weight,sex,education,type of operations,complications,preoperative TCM syndrome type,MMSE score at 1 day before operation and 7 days after operation,duration of operation,emergence time,and intraoperative blood loss,cardiovascular events,and amount of fluid infused per hour were recorded.The risk factors for POCD were analyzed using multivariate logistic regression analysis.Results Thirty-eight patients developed early POCD (25.3％).The resuhs of logistic regression analysis showed that age ≥ 70 yr,elementary education or below,duration of operation ≥5 b,and the number of hypotension occurred during operation≥3 were the risk factors for early POCD.Conclusion Age≥70 yr,elementary education or below,duration of operation ≥ 5 h,and the number of hypotension occurred during operation ≥ 3 are the risk factors for early POCD in the elderly patients undergoing spinal surgery.

Objective To evaluate biodistribution and pharmacokinetics pattern of ~(131)I-labeled rch24which is the region-grafted (humanized) anti-carcinoembryonic antigen (CEA) monoclonal antibody in nude mice. Methods Nude mice bearing cancer xenografts received intravenous injections of ~(131)I- rch24, then blood, plasma, heart, liver, spleen, lung, kidney, tumor and other tissues were taken at different time point for determination the concentration of radioactivity and calculate the T/NT value. Nude mice were packeted randomly to four group of high, medium, low dose and continuous administration, blood drug concentration was detected by ELISA method at the different intervals. Then, draw the concentration-time curve and calculate the pharmacokinetics paramete. Results After administration, radioactivity of the tumour was significantly enhanced whereas radioactivity of normal tissues decreased gradually. For single administration, at the dose of low to medium, pharmacokinetics pattern was linearity -kinetics whereas for high dose group,pharmacokinetics paramete shown some behavior of non-linearity-kinetics. Conclusion Our results suggest that the ~(131)I-labeled region-grafted (humanized) anti-CEA monoclonal antibody rch24 exhibit a considerable targeting activity so as to ~(131)I radioisotopes can be concentrated specifically in tumor. The pharmacokinetics pattern of this medicine was different at different dose.

AIM: To observe the clinical efficacy of anti-VEGF combined with laser in treatment of retinal vein occlusion with macular edema.

METHODS: Totally 62 patients with retinal vein occlusion complicated with macular edema were diagnosed in the Ophthalmology Department of our hospital during 2017-03/09 were randomly divided into three groups. Group A was treated by retinal laser alone, and groups B and C were treated with intravitreal injection of conbercept and rezuzumab respectively before retinal laser a follow-up of 6mo was conducted to observe the clinical efficacy of three groups.

RESULTS: After treatment, the retinal thickness(CMT)in macular center of the three groups was significantly lower than that before treatment, and the CMT of groups B and C were lower than that of group A(P<0.05), but there was no difference between the two groups(P>0.05). At 6mo after treatment, the total effective rate of the three groups was 44%, 86% and 86%, respectively(P<0.05), and the total effective rate of groups B and C was higher than that of group A(P<0.0167).

CONCLUSION: Vitreous injection of conbercept or ranibizumab combined with retinal laser in treatment of retinal vein occlusion with macular edema can effectively reduce macular edema and improve vision. Conbercept or ranibizumab has similar effects.

Objective To observe the effects of long-term hemodialysis(HD) combined with hemoperfusion(HP) on the levels of protein-bound uremic toxins (PBUTs) in maintenance hemodialysis (MHD) patients.Methods Forty-six patients with MHD were selected and divided into HD +HP group and HD group .HD+HP group ( n=22 ) was treated with low-flux HD twice a week and HD combined with HP once a week ,while HD group(n=24) was treated with low-flux HD three times a week.The follow-up lasted 36 weeks.The pre-dialysis concentration of PBUTs was measured at week 12, 24, 36 and baseline.PBUTs included hippuric acid (HA), indoxyl sulphate (IS)and p-cresyl sulphate (PCS).High performance liquid chromatography-tandem mass spectrometry ( HPLC-MS/MS) was used for determination .Results After 36 weeks of follow-up, the concentration of the three toxins in the HD +HP group was lower than that in the HD group during the study.At the end of the study, the reduction rates of HA, IS and PCS were 33.5%,12.8% and 24.2%, respectively, in HD+HP group.The three toxins in HD group increased by 2.3%,21.8%and 2.8%.The clearance rate of HA, PCS and IS in the HP+HD group was higher than in HD group (P<0.05).Conclusion Long-term HD combined with HP can more effectively remove PBUTs , and keep them at a lower level .

AIM: To observe application of underwater bubble method capsulorhexis overmature period to improve the small incision cataract surgery, so as to explore the clinical value of the surgical method.
● METHODS: From Jul. 2012 to Mar. 2016 at the grassroots of blindness 58 people fail in the 66 eyes overmature period of cataract were randomly divided into underwent capsulorhexis by underwater bubble method to improve the small incision cataract surgery group ( 36 eyes of 30 cases ) and conventional viscoelastic agent underwent capsulorhexis small incision cataract surgery group (30 eyes of 28 cases).
● RESULTS: A total of 66 eyes in success rate of continuous circular capsulorhexis: 92% ( 33/36 eyes ) of underwater bubble method, method of viscoelastic agent only 40% ( 12/30 eyes ) . Two groups of cases of postoperative corneal endothelial cell density are compared with preoperative significantly reduced, no significant statistical difference between the two groups(P>0. 05).
● CONCLUSION: Underwater bubble method capsulorhexis difficult to overmature period of cataract surgery capsulorhexis solution is a better way.

OBJECTIVETo explore the effect of a self-made guiding needle of steel wire in guiding the wire through the tibial tunnel for the treatment of avulsion fractures of tibial posterior cruciate ligament with open reduction and wire fixation.

METHODSFrom February 2011 to June 2014, a total of 22 patients with avulsion fractures of tibial posterior cruciate ligament underwent surgical treatments were analyzed, including 14 males and 8 females with an average age of 35.6 years old (ranged, 17 to 63 years old). According to Meyers classification, 9 patients were classified as type II, 13 patients were classified as type III. All the patients underwent open reduction and wire fixation with medial knee "L" shape approach. A wire guiding needle was used to guide the wire through the tibial tunnel during operation.

RESULTSWith the assistance of wire guidance needles, wires passed through the tibial tunnel rapidly during the operation in all the 22 patients. All the patients were followed up, X-ray imagings 6 months after operation showed the fractures healed well. The average follow-up time in all patients was 6 months (ranged, 6 to 12 months). The averaged Lysholm knee score in 22 knee was 92.7 +/- 3.4. All patients' posterior drawer test were negative.

CONCLUSIONSelf-made wire guiding needle can simplify the operation procedures in which the wires pass through the tibial tunnel, shorten the operation time, reduce the surgical trauma and complications, and be worthy of clinical application.

Adolescent ; Adult ; Bone Wires ; Female ; Follow-Up Studies ; Fracture Fixation, Internal ; Humans ; Male ; Middle Aged ; Posterior Cruciate Ligament ; injuries ; surgery ; Tibia ; injuries ; surgery ; Tibial Fractures ; surgery ; Young Adult

The formation of antisperm antibodies (AsAb) results from the disruption of the blood-testis barrier by a variety of mechanisms, which leads to exposure of immunogenic sperm antigens to the immune system and initiates an immune response. AsAb can impair the fusion of sperm and egg and even the embryo development, resulting in infertility. The etiology of AsAb, effect of AsAb on assisted reproduction and treatment of AsAb in the literature are reviewed in this article.
Antibodies ; immunology ; Antibody Formation ; Humans ; Infertility, Male ; etiology ; Male ; Reproductive Techniques ; Spermatozoa ; immunology