AIM: To establish a HPLC method for the determination of schisandrin、 deoxy-schisandrin and ?-schisandrin in Jiangmeiling Capsule(extract of Fructus Schisandrae Chinensis). METHODS: The determination was performed by RP-HPLC on Kromasil TM C 18 column(200mm?4.6mm, 5?m) using methanol-H 2O (75∶25) as a mobile phase, flow rate at 1.0mL?min -1, detection wavelength at 224nm. RESULTS: The linear range of schisandrin was 0.02228～0.24508?g,r=0.9996. The average recovery was 101.87%, RSD=1.37% (n=5). The linear range of deoxyschisandrin was 0. 02188～0. 24068?g, r=0.9997. The average recovery was 99.75%, RSD=0.94% (n=5). The linear range of ?-schisandrin was 0.01975～0.2172?g, r=0.9996. The average recovery was 100.90%, RSD=0.99% (n=5). CONCLUSION: The method is convenient, sensitive and accurate. It can be a method for quality control in production of Jiangmeiling capsule.

Objective To evaluate the in vitro effect on tumor cell uptake,tumor imaging and in vivo biodistribution of 99Tcm-epidermal growth factor receptor (EGFR) mRNA antisense PNA probe mediated by cationic liposome.Methods The oligonucleotide with sequence complementary to part of the EGFR mRNA antisense PNA was hybridized in an anti-parallel orientation targeted PNA.PNA hybridization complexes were labeled with 99Tcm by ligand exchange.The assembly of lipofectamine and 99Tcm-labeled heteroduplex was achieved by electrostatic interactions,and the radiolabeled purity was determined by reversedphase HPLC (RP-HPLC).The disparities of cell uptake in SKOV3 cells and the differences of biodistribution and molecular imaging in BALB/c nude mice bearing SKOV3 xenografts between lipofectanine-mediated 99Tcm-EGFR mRNA antisense PNA (group 1) and 99Tcm-EGFR mRNA antisense PNA (group 2) were analyzed.Two-sample t (or t') test and Wilcoxon rank sum test were used for statistical analysis.Results The labeling rates of both group 1 and group 2 were more than 95％ within 6 h.The cell uptake at 1,2,4,6,12,24 h after injection was (28.90±1.12)％,(32.76±1.20)％,(38.20±3.11)％,(41.23±1.60)％,(46.63±1.55)％ and (46.78±2.14)％ in group 1,and was (3.51±0.39)％,(3.90±0.40)％,(4.69±0.18)％,(5.91±0.26)％,(5.30±0.22)％ and (5.39±0.17)％ in group 2 respectively (t'=47.11-58.67,Z=2.80,all P＜0.05).The retention ratios showed significant difference between the two groups (t'=7.25-11.55,Z=2.80,all P＜0.05).The SKOV3 tumor could be visualized in both groups at 1 h post injection but much better visualized in group 1.The T/NT ratios were higher in group 1 at all time points (t =3.96,t'=12.65-14.69,Z=2.83-5.29,all P＜0.05).The T/NT ratios at uptake peak were 5.02 and 3.95,respectively.The probe accumulated mainly in tumor,kidneys and liver.Tumor uptake increased with time ((1.49±0.09) ％ID/g and (2.15±0.21) ％ID/g at 1 h,(3.90±0.65) ％ID/g and (5.00±0.10) ％ID/g at 6 h) after lipofectamine treatment.The ratios of tumor to contralateral muscle were also higher in group 1 (t =11.24,t' =3.96-11.94,all P＜0.05).Conclusions Lipofectamine-mediation can significantly improve the intracellular delivery of radionuclide molecular probe.Lipofectamine-mediated 99Tcm-EGFR mRNA antisense PNA can greatly improve imaging contrast and visualization of EGFR-over-expressing tumors.

Objective To prepare the 99Tcm-labeled human epidermal growth factor receptor type 2 (HER2) affibody molecule ZHER2:342 and evaluate its receptor binding specificity in vitro.Methods The molecular ZHERa:342 was labeled with 99Tcm using the ligand exchange method.The labeling efficiency and radiochemical purity were measured by HPLC.The major factors,such as the mass of SnC12 and NaOH and reaction time were analyzed,and the optimal method was summarized.Cell binding kinetics and cellular retention of the probe were investigated in HER2-expressing SKOV-3 cells and MDA-MB-231 cells with low HER2 expression respectively.HER2 binding specificity of 99Tcm-ZHER2:342 was analyzed by a pre-injection of excess unlabeled ZHER2:342 to saturate HER2 receptors.One-way analysis of variance and two-sample t test were used.Results The optimal labeling procedure was as follows:5 μg (1 g/L) of ZHER2:342 was mixed with 5 μg of NaOH (1 g/L),then 8.8 μg SnC12(1 g/L,solution) was added,followed by 150 μl (37 MBq) 99TcmO4-solution,and finally the mixture was slightly vortexed and incubated for 1 h at room temperature.99TcmZHER2:342 was stable in vitro with a high labeling efficiency of (98.10± 1.73)％.The radiochemical purity was ＞ 98％,and was more than 85％ after the incubation for 24 h in saline and fresh human serum.The cell binding of 99Tcm-ZHER2:342 with HER2-expressing SKOV-3 cells gradually increased over time with a peak of (9.95± 1.02)％ at 6 h.The binding of 99Tcm-ZHER2:342 in SKOV-3 cells was significantly higher than that in MDA-MB-231 cells at every time point (5.68-9.88 vs 0.56-2.11 ; t:from-34.50 to-13.14,all P＜0.01).The labeled molecular probe retained the capacity to bind specifically to HER2-expressing SKOV-3 cells since the cell binding decreased from (9.95 ± 1.02) ％ to (2.11 ±0.27) ％ after receptor saturation (t =-13.14,P＜0.01).Conclusions 99Tcm-ZHER2:342 has a high labeling efficiency,good stability and optimal binding specificity.These characteristics enable it to be a promising molecular probe for HER2-targeting imaging.

Objective To explore the correlation between the glucose variability and the severity of acute isolated traumatic brain injury(TBI). Method According to the inclusion/exclusion criteria,a total of 125 cases of acute isolated TBI admitted in Department of Neurosurgery of China Medical University from July 2012 to June 2015 were included. According to Glasgow coma scale(GSC),the patients were divided into five groups including control(GCS 15),mild(GCS 13?14),moderate(GCS 9?12),severe(GCS 6?8),and extra?severe(GCS 3?5)groups. Blood glucose control(including relief of the stress and the application of insulin)were carried out immediately. The average,standard deviation,and variation co?efficient of blood glucose of all groups were recorded at admission,48 hours and 3?7 days after hospitalization. The clinical records and glycemic in?dex were compared among different groups and during different periods,so as to analyze the relationship of the variability of glucose and the duration of hyperglycemia with the severity of TBI and the effects of glycemic intensive care management. Results The results of Kruskal Wallis test and Mann?Whitney Utest showed that the average,standard deviation,and variation coefficient of glucose in the extra?severe group and the severe group were statistically higher than those in the control group(P<0.05)during the same period. Meanwhile,the average,standard deviation,and variation coefficient of glucose at admission,48 hours and 3?7 days after hospitalization were also different among each group(P<0.05). The duration of hy?perglycemia and conscious disturbance in both the extra?severe group and the severe group were longer than those in the control(P<0.05). The analyses using rank correlation indicated that glucose variability,the level and duration of hyperglycemia were positively correlated with the severity of TBI(r>1). Conclusion The glucose variability in acute isolated TBI patients could be considered as the index of the severity of TBI.

There are abundant and stable circular RNAs in tumor-derived exosomes, which play an important role in the occurrences and developments of tumors. Exosome-circRNAs can be used as the marker of tumors diagnosis and prognosis, and have different roles in regulating tumors immune responses, regulating tumors progressions and mediating tumors drug resistances in different tumors. Exploring and applying the potential value of exosome-circRNAs will provide a new choice for the diagnosis and treatment of tumors.

Objective:To screen highly expressed inflammatory factors in diabetic nephropathy models using protein microarray, analyze differential genes and their regulatory networks, and predict potential therapeutic small molecular compounds.Methods:The inflammatory factor microarray was used to screen the inflammatory factors with the same tendency in the cell model and animal model of diabetic nephropathy. The differential genes screened by R language were enriched and analyzed by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes(KEGG). STRING builds a protein interaction network online, Cytoscape software analyzes the core subnetwork, and Connectivity Map searches for and predicts small molecule compounds.Results:Diabetic nephropathy model was established using 16-week-old db/db mice and mesangial cells stimulated with high glucose, and the expression of C-X-C motif chemokine ligand 1(CXCL1) was elevated in both models. Multiple GEO datasets indicated a strong association between the high expression of CXCL1 and diabetic nephropathy. Specifically, GSE30122 showed an upregulation of 30 genes and a downregulation of 23 genes. GO enrichment analysis focused on biological processes such as humoral immunity and lipopolysaccharide response; While KEGG enrichment was mainly in pertussis and coagulation cascade pathways. CytoHubba identified 10 hub genes, such as ALB, LUM, and CXCL1. In addition, 10 small molecule compounds were predicted as potential therapeutic drugs using Connectivity Map.Conclusions:CXCL1 may serve as a key gene in the occurrence and development of diabetic nephropathy. ALB, LUM, CXCL1, MMP7, TGFBI, CCL2, S100A4, SOX9, VCAN, and CLU may participate in the regulatory network centered around CXCL1. There are 10 small molecular compounds demenestrating the potential to be therapeutic agents.

Objective:To investigate the efficacy and safety of the modified gasless unilateral axillary approach (MGUAA) endoscopic thyroid surgery in treatment of papillary thyroid microcarcinoma (PTMC) .Methods:From Jan. 2019 to Dec. 2019, 90 patients receiving PTMC (cT1N0M0, cI stage, 8th, 2017 AJCC) therapy by modified gasless unilateral axillary approach endoscopic thyroid surgery (MGUAA group, n=41) , and conventional open thyroid surgery (OS group, n=49) were retrospectively analyzed. Ninety patients were enrolled in the study, including 14 males and 76 females,with the mean age (42.1±12.0) years.The effectiveness of central lymph node dissection (CLND) , the operation time, the types of operation, the amount of drainage, the duration of hospital stay, the related complications, the postoperative pain of neck and axillary and the cosmetic satisfaction were compared between the two groups.SPSS 25.0 statistical software was used for statistical analysis, the measurement data was expressed by ±s, paired t test was used to compare the measurement data between groups, and Chi-square test was used to campare the count date between groups. Results:The mean age (35.0±8.6) years and the amount of surgical bleeding (12.3±7.3) ml in the MGUAA group were significantly lower than those (48.1±11.1) years and (16.1±4.3) ml in the OS group ( P<0.01) , while the mean operation time (99.1±19.5) min, the mean amount of drainage (221.4±67.9) ml and the postoperative drainage tube placement time (5.0±0.8) days were significantly higher than those of (70.6±17.8) min, (98.3±63.7) ml and (3.8±1.0) days in the MGUAA ( P<0.01) . There was no significant difference in the number of lymph nodes of CLND or the duration of hospital stay between the two groups ( P>0.05) . In terms of surgical complications, the transient recurrent laryngeal nerve injury, the postoperative hematoma, the postoperative infection, and the lymphatic leakage had no significant difference between the two groups ( P>0.05) . The MGUAA group had significant advantages in avoiding the postoperative dysphagia in front of neck, the postoperative pain of neck, and cosmetic satisfaction over the OS group [ (0.0% vs 28.6%) , (14.6% vs 71.4%) , (1.1±0.3) score vs (2.4±0.5) score ( P<0.01) ]. Whereas in axillary area pain on the surgical side, the MGUAA group was inferior to the OS group ( P<0.01) . Conclusion:The modified gasless unilateral axillary approach endoscopic thyroid surgery is a feasible, safe and cosmetically operation for PTMC (cT1N0M0, cI stage, 8th, 2017 AJCC) .

Programmed death-1 (PD-1) and programmed death ligand-1 (PD-L1) are involved in the regulation of immune checkpoints and are closely related to the occurrence and development of tumors. In thyroid cancer, an increase in PD-L1 expression and an increase in PD-1 positive T cells may be predictive of higher invasiveness and a greater risk of recurrence. Anti-PD-1/PD-L1 therapy has had significant effects in many tumors, but the efficacy in thyroid cancer is still relatively limited, which also requires finding biomarkers those can better predict the efficacy. Further understanding of the mechanism of PD-1/PD-L1, the current research status in thyroid cancer, and biomarkers related to its efficacy may provide new treatment methods and ideas for patients with thyroid cancer.

OBJECTIVE: To explore the regulatory mechanism of human hepatocyte apoptosis induced by lysosomal membrane protein Sidt2 knockout. METHODS: The Sidt2 knockout (Sidt2^-/-) cell model was constructed in human hepatocyte HL7702 cells using Crispr-Cas9 technology.The protein levels of Sidt2 and key autophagy proteins LC3-II/I and P62 in the cell model were detected using Western blotting, and the formation of autophagosomes was observed with MDC staining.EdU incorporation assay and flow cytometry were performed to observe the effect of Sidt2 knockout on cell proliferation and apoptosis.The effect of chloroquine at the saturating concentration on autophagic flux, proliferation and apoptosis of Sidt2 knockout cells were observed. RESULTS: Sidt2^-/- HL7702 cells were successfully constructed.Sidt2 knockout significantly inhibited the proliferation and increased apoptosis of the cells, causing also increased protein expressions of LC3-II/I and P62(P < 0.05) and increased number of autophagosomes.Autophagy of the cells reached a saturated state following treatment with 50 μmol/L chloroquine, and at this concentration, chloroquine significantly increased the expressions of LC3B and P62 in Sidt2^-/- HL7702 cells. CONCLUSION Sidt2 gene knockout causes dysregulation of the autophagy pathway and induces apoptosis of HL7702 cells, and the latter effect is not mediated by inhibiting the autophagy-lysosomal pathway.
Humans ; Lysosome-Associated Membrane Glycoproteins/metabolism* ; Autophagy ; Apoptosis ; Hepatocytes ; Lysosomes/metabolism* ; Chloroquine/pharmacology* ; Nucleotide Transport Proteins/metabolism*