Objective To investigate the kinetics of absorbing timosaponin by AB-8 resin.Methods Static absorption experiment,dynamic absorption experiment and series dynamic absorption experiment were carried out,and then the content of timosaponin was detected by spectrophotometry.Finally,the static absorption curve and the dynamic absorption curve were figured out.Results The result of static absorption experiment showed that the parameters of Lagergren equation are Kad=0.0186,r=0.9986,and the parameters of Dumwals-Wagner are K=0.0081,r=0.9958.In dynamic absorption experiment,the leaking of timosaponin is found and the penetrating point is at 600mL(0.1g/mL).Conclusion The mechanism of AB-8 resin absorbing timosaponin is characterized as liquid membrane diffusion at the early phase and intragranular diffusion at the later phase.Leaking phenomena in dynamic absorption experiment can be solved by three tubes in series.Therefore,this method can be used for the research of Chinese herbal medicine.

NS2 is a nonstructural protein of Periplaneta fuliginosa densovirus(PfDNV) with a molecular mass of 30 kD,whose function is not yet clearly understood. In order to study the expression,subcellular distribution and the function of NS2 protein,the coding region of NS2 was amplified from the hindgut tissue of cockroaches infected with PfDNV by RT-PCR and then the recombinant prokaryotic expression vector pET28a-NS2 was constructed. The recombinant plasmid was transformed into E. coli BL21(DE3) to express the 6?His fusion protein in the bacteria. After purification,the fusion protein was injected into New Zealand rabbits to prepare polyclonal antibody. The specificity of the anti-NS2 antibody was successfullyproved by western blotting on the eukaryotic expressed products of NS2 protein.Meanwhile,the full sequence of ns2 gene was also cloned into the eukaryotic expression vector pAC. The recombinant plasmid pAC-NS2 was then transfected into Schneider line 2(S2) cells to express NS2 protein in the insect cells. The subcellular localization of NS2 in the insect cells was then investigated by indirect immunofluorescence technique using the anti-NS2 polyclonal antiserum. The confocal laser scanning microscope observation showed that NS2 protein was located primarily in the cytoplasm with some punctate nuclear staining.

Objective To develop a RP-HPLC method for the determination of piperine in the root of Piper nigrum L.Methods RP-HPLC was carried out on Luna C18 column(250 mm? 4.60 mm,5 ? m) with the column temperature of 35 ℃.The mobile phase consisted of a mixture of methanol-water(77 :23) at a flow rate of 1.0 mL? min-1.The determination wavelength was at 343 nm.Results The calibration curve was linear within the concentration range of 0.164 ? g～ 0.984 ? g,r=0.9996,and the average recovery was 98.09 %,RSD=2.67 %(n=9).The average content of piperine in three batches of pepper roots was in the range of 6.67～6.77mg?g-1.Conclusion Pepper root contains piperine,and this method is suitable for the quality control of the root of Piper nigrum L.

Objective To establish the quality standard for Bazheng Pills. Methods Radix et Rhizoma Rhei, Radix Glycyrrhizae and Fructus Gardeniae in Bazheng Pills were identified by TLC. Geniposide was determined by HPLC. Results Radix et Rhizoma Rhei, Radix Glycyrrhizae and Fructus Gardeniae could be identified by TLC. Geniposide showed a good linear relationship at a range of 0.187 2～ 2.808 0 ng, r=0.999 9.The average recovery was 100.70 % , and RSD was 1.01 % . Conclusion The established methods are simple and with good reproducibility, and can be used for the quality control of Bazheng Pills.

Objective: To prepare anti-VSIG4 monoclonal antibodies and characterize their biological functions.Methods: BALB/c mice were immunized with transfected cell line (L929/VSIG4L) as immunogen.The spleen B cells of the mice were fused with SP2/0 and hybridoma cells were screened with transfected cell line (L929/VSIG4) by FCM.After acquisition of the hybridomas secreting anti-VSIG4 mAb,their biological activities were investigated by indirect immunofluorescence,Western blot,competitive inhibition test,and MTT assay.Results:Two stable hybridomas,9A7 and 9D5 were obtained,which could continuously secrete specific anti-VSIG4 monoclonal antibodies.The following biological activity studies showed that these monoclonal antibodies could recognize the natural VSIG4 expressed on the macrophages and several cancer cell lines,such as Jurkat,THP-1 and H446.Furthermore,they could block the inhibitory effects of VSIG4 on proliferation of T cells in vitro.Conclusion: Two hybridomas secreting anti-VSIG4 monoclonal antibodies have been established.These monoclonal antibodies provide useful tools for further studying VSIG4's biological function and its unknown receptor.

Objective:To investigate the effect of recombinant lipoproteins of Brucella outer membrane protein 16, 19 (L16 and L19) on the expression of immune regulatory factors in human monocytic leukemia cell line (THP-1 cells). Methods:THP-1 cells activated with phorbol ester (PMA) were used as an in vitro experimental cell model, and a group design was used to co-culture L16, L19 and THP-1 cells (L16 stimulated group, L19 stimulated group), respectively. THP-1 cells activated with PMA were used as the control group. When co-cultured for 4 hours, immunofluorescence staining (IFS) and Western blotting were used to detect whether L16 and L19 entered the cells, respectively; when co-cultured for 12, 24 hours, real-time fluorescent quantitative PCR was used to measure the mRNA expression levels of interferon regulatory factor 1 (IRF-1) and trans activator protein of major histocompatibility complex class Ⅱ (CⅡTA); Western blotting was used to detect the protein expression levels of T cell immunoglobulin mucin-3 (Tim-3) and γ interferon receptor 1 (IFNGR1). Results:When co-cultured for 4 hours, L16 and L19 were observed entering THP-1 cells in the L16 stimulated group and L19 stimulated group, respectively. When co-cultured for 12 hours, the expression level of IRF-1 mRNA in the L16 stimulated group (0.16 ± 0.15) was significantly lower than that in the control group (1.00 ± 0.00, P < 0.05). When co-cultured for 24 hours, the expression level of CⅡTA mRNA in the L16 stimulated group (0.17 ± 0.10) was significantly lower than that in the control group (1.00 ± 0.00, P < 0.05). When co-cultured for 12 and 24 hours, there were no statistically significant differences in the expression levels of IRF-1 and CⅡTA mRNA between the L19 stimulated group and the control group ( P > 0.05). Western blotting results showed that there were statistically significant differences in the expression levels of INFGR1 and Tim-3 protein among the control group, L16 stimulated group, and L19 stimulated group after co-cultured for 12 and 24 hours ( F = 50.92, 6.80, 148.73, 156.57, P < 0.05). Among them, when co-cultured for 12 hours, the expression level of INFGR1 protein in the L16 and L19 stimulated groups were significantly lower than that in the control group, and the L19 stimulated group was higher than the L16 stimulated group ( P < 0.05), and the expression level of Tim-3 protein in the L19 stimulation group was higher than that in the control group ( P < 0.05). When co-cultured for 24 hours, the expression level of INFGR1 protein in the L16 and L19 stimulated groups were lower than that in the control group, and the L19 stimulated group was higher than that in the L16 stimulated group ( P < 0.05); and the expression level of Tim-3 protein in the L16 stimulated group was higher than that in the control group and L19 stimulated group ( P < 0.05). Conclusions:Brucella L16 can downregulate the expression levels of IRF-1 and CⅡTA mRNA in THP-1 cells. Both L16 and L19 can downregulate IFNGR1 and upregulate Tim-3 protein expression levels.

Objective:To explore the risk factors for myelosuppression in elderly lung cancer patients undergoing chemotherapy and construct a risk prediction model for myelosuppression in elderly lung cancer patients undergoing chemotherapy.Methods:Using the convenient sampling method, data of 228 elderly patients with lung cancer undergoing chemotherapy in Respiratory Department of a Class Ⅲ Grade A hospital in Zhenjiang from May 2018 to May 2019 were selected, and risk factors of adverse reactions of myelosuppression in patients were analyzed statistically. The binomial Logistic regression was applied to construct the prediction model and the area under the ROC curve was used to test the prediction effect of the model. The patient data from January to May 2020 were collected to validate the model.Results:Among the 228 patients, 75 patients developed myelosuppression, with an incidence of 32.89%. Multivariate analysis results showed that platinum-containing chemotherapy regimens, combined with other adverse reactions, decreased albumin before chemotherapy and decreased hemoglobin before chemotherapy were independent risk factors for myelosuppression in elderly lung cancer patients during chemotherapy ( P<0.05) , which were included in the model. The area under the ROC curve of the final model was 0.823, the maximum Youden index was 0.5, sensitivity was 81.3%, and specificity was 70.5%. The results of the verification data showed that the area under the ROC curve was 0.846, sensitivity was 90.4% and specificity was 68.2%. Conclusions:The prediction effect of this model is good, which can provide reference basis for clinical treatment and formulating nursing measures to prevent myelosuppression.

Objective:To understand the current situation of postpartum maternal perceived spouse support for breastfeeding, and explore its mediating role in the spouse support for breastfeeding and maternal breastfeeding self-efficacy.Methods:From October 2021 to October 2022, convenience sampling was used to select 602 postpartum women (42 days postpartum) and their spouses who underwent follow-up visits at Obstetric Clinics in four ClassⅢ hospitals in Shanghai as the research subject. A questionnaire survey was conducted using the General Information Questionnaire, Partner Breastfeeding Influence Scale (PBIS) , and Breastfeeding Self-efficacy Scale Short Form (BSES-SF) .Results:The PBIS score of 602 postpartum women was (121.07±25.41) , and the PBIS score of 602 maternal spouses was (134.37±18.94) . Pearson correlation analysis results showed that maternal perceived spouse support has a positive correlation with spouse support ( P<0.01) , and a positive correlation with maternal self-efficacy in breastfeeding ( P<0.01) ; spouse support was positively correlated with maternal self-efficacy in breastfeeding ( P<0.01) . The mediating effect analysis that maternal perceived spouse support played a complete mediating role between spouse support and maternal self-efficacy in breastfeeding. Conclusions:There are both connections and differences between maternal perceived spouse support and spouse support, and the positive promoting effect of spouse support on maternal self-efficacy in breastfeeding can be completely mediated by maternal perception. Medical and nursing staff should pay attention to the assessment of maternal perceived spouse support and guide spouse breastfeeding support behavior based on maternal needs and experiences of spouse support, so as to improve breastfeeding outcomes.

Objective:This study is designed to investigate the toxicity of lipoprotein (L16) and non-lipoprotein (U16) of Brucella outer membrane protein (OMP) 16 on osteoblasts. Methods:Recombinant L16 and U16 proteins were prepared by using prokaryotic expression system of Escherichia coli ( E. coli) BL21 (DE3) and purified by Ni column. Using group design, mouse osteoblasts (MC3T3 cells) were co-incubated with L16 and U16, respectively. Brucella lipopolysaccharide (LPS) stimulus was used as the positive control, and cells without any stimulation were used as the negative control. Incubation time was 24 h. The activity of co-incubated MC3T3 cells were detected by CCK-8; the supernatant of cultured cells was collected and the release rate of lactate dehydrogenase (LDH) in the supernatant was detected by bioluminescence, and the virulence of L16 and U16 on MC3T3 cells was evaluated. Annexin Ⅴ-PE/7-AAD double staining flow cytometry was further used to analyze the apoptosis rate of MC3T3 cells, and the activation level of apoptosis executive protein Caspase-3 was detected by Western blotting (WB). Results:The activity of MC3T3 cells in L16 group [(56.16±1.63)%] was significantly lower than that in U16 and LPS groups [(97.02±1.44)%, (98.64±0.90)%, P < 0.01], the LDH release rate [(84.64±0.96)%] was significantly higher than that in U16 and LPS groups [(34.82±3.41)%, (26.75±1.95)%, P < 0.01]. Annexin Ⅴ-PE/7-AAD double staining results showed that the apoptosis rate was (46.45±2.19)% in L16 group, while the remaining groups were all less than 1%. WB results showed that activated Caspase-3 (cleaved-Caspase-3) existed in L16 stimulated cells, but not in U16 stimulated cells and LPS control cells. Conclusion:L16 can induce the apoptosis of osteoblasts and inhibit the proliferation of osteoblasts, but U16 has no obvious effect indicating that Brucella L16 with complete lipid structure is necessary for virulence effect.