To investigate the immunoregulatory effect of Lycium barbarum polysaccharides (LBP) of different molecular weights (Mw) on RAW264.7 macrophages.

LBP with different Mw was obtained by water extraction, alcohol precipitation and chromatographic column separation. CCK8 method was used to detect the effect of LBP on proliferation of RAW 264.7 cells. The effect of LBP on cell phagocytosis was determined by neutral red method. Real-time quantitative PCR was applied to detect the mRNA expression of TNF-α, IL-6, IL-1β and iNOS. Western blot was used to detect the phosphorylation level of proteins in MAPK signaling pathway.

Two high purity homogeneous components, LBPl (Mw=1 207 kDa) and LBP2 (Mw=125 kDa) were obtained by separation and purification. Compared with the control group, LBP1 and LBP2 at different concentrations significantly promoted the proliferation and phagocytosis of macrophages (P<0.05). LBP1 and LBP2 in the concentration range of 50~200 μg·mL^-1 could promote the expression of TNF-α, IL-1β, IL-6 and iNOS mRNA in macrophages in a dose-dependent manner. Western blot analysis showed that LBP1 and LBP2 induced immune response in RAW264.7 cells, which was related to the specific activation of JNK MAPK pathway. In addition, the regulatory effect of LBP1 was better than that of LBP2 on RAW264.7 cells.

The immunoregulatory effect of LBP on RAW264.7 macrophages may be related to its specific activation of JNK MAPK signal transduction pathway, and its activity is closely related to the Mw of LBP.

To establish an indirect enzyme-linked immunosorbent assay (ELISA) for determination of MBS77E, a novel monoclonal antibody against Ebola virus, in the serum of cynomolgus monkeys and to study its pharmacokinetics in monkeys.

An indirect ELISA method for quantitative detection of MBS77E in cynomolgus monkey serum was developed and fully validated. Four cynomolgus monkeys were divided into two groups. Each group of monkeys were administered with MBS77E via intravenous injection at dosage of 5 or 100 mg·kg^-1, respectively. Blood samples were collected at different time points after administration. Serum was obtained and the concentration of MBS77E was determined using the established ELISA method. The pharmacokinetic parameters of MBS77E were analyzed using WinNonlin software.

The linear range of the method was 0.098~12.5 μg·mL^-1. The precision RSDs of the intra- and inter-assay were within 3.5%~9.9% and 7.4%~11.8%, respectively. After i.v. injection at dosage of 5 and 100 mg·kg^-1, the terminal t_1/2 of MBS77E was (8.88±5.24) and (6.38±2.57) d, CL was (7.55±1.48) and (6.22±0.99) mL·h^-1·kg^-1, V was (91.18±38.16) and (55.44±14.01) mL·kg^-1, AUC was (631.58±76.75) and (16 004.13±2 230.73) μg·d·mL^-1, respectively. The AUCs were proportional to the two dosages. The plasma clearances of MBS77E did not change with different dosages.

An indirect ELISA method was successfully established for the determination of MBS77E in monkey serum which met the requirements of preclinical pharmacokinetic research in terms of specificity, linear range, precision and stability et al. The pharmacokinetic characteristics of MBS77E in cynomolgus monkeys were obtained. This study provided support for the preclinical and clinical pharmacokinetic studies of MBS77E.

To analyze the follow-up changes of subjects of anti-cancer clinical trials in our department under the COVID-19 epidemic situation.

The clinical trials were classified. The follow-up status of subjects in our department from February 3 to April 30 and May 6 to 29, 2020 was collected, and the data were input and analyzed by Excel and SPSS 22.0.

There were 32 anti-cancer clinical trials in our department. From February 3 to April 30, 2020, 1 049 person times were planned to be followed up, and 658 person times were actually followed up, accounting for 62.73%. The follow-up rate of oral drugs was 93.33%, that of oral drugs combined with non-oral drugs was 87.73%, and that of non-oral drugs was 70.71%. In May 2020, the actual follow-up rate of subjects on site rose to 88.97%, while the number of delayed drug users, the number of external hospital inspectors and the number of non-drug users decreased.

The epidemic situation had a great impact on the follow-up of anti-cancer clinical trials in a short time, but did not cause a sustained impact. The establishment of relevant follow-up measures improved the follow-up rate of subjects to some extent. For the affected subject population, further analysis is needed to better guide the follow-up work of subjects under emergency.

To investigate the protective effect of gardenoside on dementia-like mice induced by tail intravenous injection of streptozotocin (STZ).

Mice receiving STZ tail intravenous injection were randomly divided into model group (Model), low-dose gardenoside group (GL), high-dose gardenoside group (GH), and solvent control group (Vehicle). Blood glucose level was monitored every two weeks. After administration of the drug, a glucose tolerance test was conducted, and serum insulin levels were detected by ELASA. Morris water maze test was used to test the learning and memory ability of mice in each group, and Contextual fear system was used to test the ability of fear memory regression of mice in each group. Western blot was used to detect the expression of GSK-3β and phosphorylated GSK-3β (P-GSK3β) protein in hippocampus and cortical tissues.

Compared with the Vehicle group, the blood glucose level in the Model group was significantly increased (P<0.001). The spatial learning and memory function as shown by Morris water maze detection were significantly impaired (P<0.05). In the episodic fear memory experiment, there were disorder in the extinction of fear memory in mice (P<0.01) and significant decrease in serum insulin (P<0.05), while serum insulin resistance index increased significantly (P<0.001). The expression level of GSK3β remained unchanged in hippocampus and cortex, while the expression level of P-GSK3β decreased significantly (P<0.05). All abnormal indexes in the GH group were improve (P<0.05). In the GL group, some abnormal indexes were improved to a certain extent.

Gardenoside can improve spatial learning and memory impairment and fear memory extinction disorders in mice receiving STZ tail intravenous injection. The mechanism may be increasing P-GSK3β to inhibit GSK-3β activation, thereby protecting nerve cells.

To identify the related substances in mesaconine.

An HPLC-ELSD method was established using Gemini C₁₈ column (150 mm×4.6 mm, 5 μm) with mobile phase composed of methanol-0.1% trifluoroacetic acid to separate the impurities. Forced degradation tests were designed and performed on mesaconine to specify the impurities. Ultra-high performance liquid chromatography tandem time-of-flight mass spectrometry (UPLC-TOF/MS) and triple quadrupole mass spectrometry (UPLC-MS/MS) were used for determination of exact mass and product ions of impurities. The structures of impurities and fragmentation regularity were then figured out.

A total of four impurities were detected and identified in mesaconine, including three impurities derived from synthesis process and a degradation product.

UPLC-MS/MS technology can effectively separate and identify the impurities in mesaconine, and the results provide a valuable reference for the manufacturing process and quality control of mesaconine.

To establish a method for determination of polymer impurities in ceftazidime averbatan sodium for injection.

Ceftazidime averbatan sodium was dissolved by water to prepare polymer stock solution. High performance size exclusion chromatography method (HPSEC) and column switching-LC/MSn method were applied to separate and deduce the poor retention impurities in polymer stock solution, and the specificity of HPSEC method was evaluated. A RP-HPLC method for polymer analysis was established with C₁₈ column, using gradient elution by phosphate buffer solution (pH 3.2)-acetonitrile-methanol, and the RP-HPLC method was validated.

Five polymers were identified in polymer stock solution, some small molecular impurities were also detected. Polymer impurities were liable to co-elute with small molecular impurities when using HPSEC method, which resulted in poor quantification accuracy and specificity. Four indicative polymer impurities were detected by RP-HPLC method with effective specificity, and no polymers related to averbatan sodium were detected.

HPSEC method is not suitable for the quality control of polymer impurities in ceftazidime averbatan sodium for injection, while the newly established RP-HPLC method is specific, which can effectively control the polymer impurities. The polymer impurities in the samples were all from ceftazidime, and averbatan sodium did not produce polymer impurities.

To establish an HPLC-CAD method for determination of mannitol in somatostatin for injection.

Mannitol was determined by HPLC-CAD with Agilent 300Extend-C₁₈ column (250 mm×4.6 mm, 5 μm), the mobile phase was acetonitrile-water (15∶85) at the flow rate of 1.0 mL·min^-1, the column temperature was 35 ℃, and the injection volume was 20 μL. Charged aerosol detector (CAD) was used as detector, the evaporate tube temperature was 50 ℃, nebulizer gas was nitrogen, the flow pressure was about 60 psi, and the gain value was 1.

The detection limit for mannitol was 1.004 μg·mL^-1, and the limit of quantitation was 3.012 μg·mL^-1. The quadratic curves showed good linearity in the range of 0.1~1.0 mg·mL^-1(r²=0.999 8). The average recovery was 99.1% (RSD=0.60%).

This method is simple, rapid, sensitive, accurate and suitable for the quality control of mannitol in somatostatin for injection.

To evaluate the effectiveness, safety and economy of pembrolizumab (PEM) for locally advanced/metastatic NSCLC.

Chinese and English databases and HTA organization websites were systematically searched. Two reviewers independently identified studies, extracted data and assessed the quality of included studies. Qualitative analysis was performed from the perspectives of efficacy, safety and economy.

Three HTA reports, six systematic reviews and 14 pharmacoeconomic studies were included. In terms of effectiveness, compared with chemotherapy (CT), PEM alone could improve the overall survival (OS) of patients of locally advanced or metastatic NSCLC with PD-L1 50%, but not that of patients with PD-L1 1%~49%. PEM+CT could improve (with statistical significance, the same below) the OS and progressive-free survival (PFS) of locally advanced or metastatic NSCLC patients (regardless of PD-L1 expression level) and metastatic non squamous NSCLC patients; it could improve the PFS of metastatic squamous NSCLC patients. Compared with NIV, PEM+CT could improve the OS and PFS of PD-L1 50% of advanced or metastatic NSCLC patients and metastatic non squamous NSCLC patients, but it did not improve the OS and PFS of metastatic squamous NSCLC patients. Compared with NIV+CT, PEM+CT could improve the OS of patients with mutation negative metastatic NSCLC. Compared with BEV, PEM+CT can improve the OS of advanced or metastatic NSCLC patients with negative mutation and nonsquamous NSCLC patients with negative mutation. In terms of safety, compared with CT, PEM could reduce grade 1~5 treatment-related side effects, grade 3~5 treatment-related side effects and serious side effects in patients with advanced or metastatic NSCLC with negative mutations. Compared with CT, the incidence of grade 1~5 treatment-related side effects, grade 3~5 treatment-related side effects and serious side effects were comparable. Compared with NIV, PEM+CT increased severe side effects. The severe side effects of PEM+CT were comparable to those of NIV+CT. In terms of economy, compared with CT, for mutation-negative locally advanced or metastatic NSCLC patients with PD-L1 1%, PEM alone is not cost-effective in China. For mutation-negative metastatic non-squamous NSCLC patients, PEM+CT is also not cost-effective in China.

PEM has favorable efficacy and safety for the first-line treatment of NSCLC, However, according to its price analyzed in the study, it is not cost-effective in China currently and further price reducing may help improve its cost-effectiveness.