Objective To obtain monoclonal antibodies ( mAbs ) against neutrophil gelatinase-associated lipocalin ( NGAL ) and a chemiluminescense immune quantification assay based one paired mAbs.Methods Six-to-eight weeks old female BALB/c mice were immunized with the purified recombinant human NGAL antigen( rhNGAL) that was produced by the Escherichia coil expression system.The spleen was fused with hybridoma for screening anti-NGAL monoclonal antibodies by indirect ELISA.Western blot was implemented to identify the reactivity with native NGAL. Results The rhNGAL antigen was found to form disulfide cross-linked dimers and present excellent immunogenicity.The reaction titer of the immune serum of NGAL immunized mice was about 106.Thirty mAbs were screened by indirect ELISA, hereinto;the EC50 values of mAb23C12 and 38D10 were 0.034 g/mL, 0.022 g/mL respectively.The antibodies pair, 38D10/23C12-SAE labeled with AcridiniumEster(AE), were shown to work well in chemiluminescense immune response quantitative detection which was screened by NGAL standardand clinical urine samples.This detection can resolve positive and negative samples with a statistically significant difference (P<0.0001).And the correlation coefficient R2between NGAL quantitative results and that of the Abbott's NGAL chemiluminescence immune assay kit was greater than 0.97.The detection linear range was 10-1500 ng/mL, analytical sensitivity of the method was 0.63 ng/mL.Conclusion Highly purified rhNGAL antigen and specific anti-NGAL monoclonal antibodies are generated in this study.The detection capability of method is comparable with that of the international commercial kit.

OBJECTIVE:To optimize extraction technology of Shenqi fukang granules. METHODS:Single factor method was used to investigate water multiple at first time using water absorption as index,soaking time using yield of dry extract as index. Based on single factor test,by weighted comprehensive scoring method,using the yield of dry extract and the content of 75％ ethanol extraction as indexes,extraction time,water multiple and extraction times as factors,L9(34) orthogonal test was used to optimize water extraction technology of Shenqi fukang granules. Validation test was conducted. RESULTS:The optimal water extraction technology included 8-fold water (L/kg), extracting for 3 times, 1 h each time. CONCLUSIONS:The optimized extraction technology is stable and controllable in quality,and can provide reference for industrial production of Shenqi fukang granules.

Objective:To explore the effect of intelligent rehabilitation device on lower limb function rehabilitation in stroke patients with hemiplegia.Methods:Using convenience sampling, 186 stroke hemiplegic patients admitted to the Department of Neurology and Surgery of Brain Hospital, Cangzhou Central Hospital from July 2021 to July 2022 were selected as the study subject. The patients were divided into a control group, a strengthening group, and an intelligent rehabilitation group, with 62 patients in each group using the random number table. The control group received routine rehabilitation for 40 minutes per day, 5 days per week, for a total of 4 weeks. On the basis of the control group, the strengthening group received an additional 30 minutes of routine rehabilitation training every day. The intelligent rehabilitation group received an additional 30 minutes of intelligent rehabilitation device exercise per day on top of the control group. The Fugl-Meyer Assessment Lower Extremity (FMA-L), Berg Balance Scale (BBS) scores, and Functional Ambulation Category (FAC) grading of three groups of patients before and after intervention were compared.Results:Sixty patients in each group completed the study. The FMA-L and BBS scores of the three groups of patients after intervention were higher than those before intervention, and the FAC grading was better than that that before intervention, with statistical differences ( P<0.05). After intervention, the FAM-L and BBS scores of the intelligent rehabilitation group were higher than those of the control group and the strengthening group, and the FAC grading was better than that of the control group and the strengthening group, and the differences were statistically significant ( P<0.05) . Conclusions:The intelligent rehabilitation device can improve the lower limb function of stroke patients with hemiplegia, and is worthy of clinical promotion and practice.

Objective : To investigate the effects of RO or incubation with cholesterol ( CH) on the differentiation and apoptosis of KCs. Methods : Ca2 + ( 1. 8 mmol/L) was added to KCs for 1 day after co - culture of RO alone or combined with CH in KCs for different time. The expression of Involucrin (INV) and Loricrin in KCs was analyzed by Western blot. After co⁃culture of RO alone or combination with CH in KCs for different time , the apoptotic changes of KCs cells were detected by flow cytometry and the expression of apoptotic proteins Bax and Bcl⁃2 were verified by Western blot. Results : RO down⁃regulated the expression of Ca2 + induced differentiation marker protein INV , but weakly inhibited the expression of Loricrin , while CH showed no antagonistic effect on RO. RO induced apoptosis of KCs cells in a time dependent manner. CH could antagonize the apoptotic effect of RO on KCs ; the expression of Bcl⁃2 and Bax was inhibited when RO was applied alone , CH could partially antagonize the inhibition effect of RO on Bcl⁃2 expression , but had no significant effect on Bax; however, RO reduced the ratio of Bcl⁃2/Bax in a time⁃dependent manner, and CH partially weakened the effect of RO on the ratio of Bcl⁃2/Bax. Conclusion RO may inhibit KCs differentiation and induce KCs cell apoptosis by down⁃regulating the expression of Loricrin and Bcl⁃2.