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.

This paper introduces the historical development, training contents, the inspection mechanism and management regulations of obstetrics and gynecology residents training in Taiwan region, China. Drawing lessons from the experience and ideas of the standardized residency training system of obstetrics and gynecology in Taiwan region, combined with the present situation and deficiencies of the training system in the mainland China, this paper puts forward constructive suggestions for the standardized residency training of obstetrics and gynecology, so as to obtain more efficient and satisfactory training results and provide ideas for cultivating excellent obstetrics and gynecology specialists in mainland China.

Objective To explore the value of 3.0T MR elastography (MRE) in diagnosis of obstructive chronic pancreatitis.Methods Totally 32 patients (lesion group) with suspected obstructive chronic pancreatitis who underwent pancreaticoduodenectomy and 32 volunteers (normal control group) were enrolled.MRE was performed,and pancreatic stiffness value was measured.The consistency between two observers and the repeatability of the same observer were evaluated.The difference of pancreatic stiffness value was compared between the two groups.The efficacy of pancreatic stiffness value in diagnosis of obstructive chronic pancreatitis was analyzed with ROC curve.Results The consistency between two observers and the repeatability of the same observer were excellent (all ICC＞0.9).The pancreatic stiffness value of normal control group and lesion group was (1.21±0.11)kPa and (1.51±0.24)kPa,respectively (t=-6.077,P ＜0.001).The area under ROC curve of pancreatic stiffness value in diagnosis of chronic pancreatitis,mild and moderate to severe,mild to moderate and severe was 0.900,0.941 and 0.960,respectively (all P＜0.001).Conclusion MRE can objectively measure pancreatic stiffness and noninvasively assess the severity of chronic pancreatitis.

Objective To explore the clinical application value of prognostic nutritional index (PNI)for predicting overall survival(OS)in patients with advanced colorectal cancer.Methods Seventy-two patients with histologically confirmed colorectal cancer were enrolled in this study, and their clinical and laboratory data were reviewed.The PNI was calculated, and univariate and multivariate analysis was used to identify the potential prognostic factors for advanced colorectal cancer. Results PNI of the 72 colorectal cancer patients was 45.07 ± 5.98.PNI was significantly associated with age, weight loss and pleural effusion (P < 0.05). However, PNI showed no correlation with sex, clinical stage, smoking, bloody stool, abdominal mass, intestinal obstruction, histological type, radiotherapy and KPS scores (P>0.05).The median OS of the 72 patients was 12.9 months.The medium OS in the higher PNI group (PNI≥45.07)and lower PNI group(PNI≤45.07)was 15.7 months and 11.2 months, respectively.The 1-year survival rates were 78.4% and 60.5%, and 2-year survival rates were 53.1% and 20.8%, respectively(P<0.01).Univariate analysis showed that PNI, ages, and weight loss were related to the OS of the advanced colorectal cancer(P<0.05).Multivariate analysis identified PNI was an independent prognostic factor for OS of advanced colorectal cancer (P < 0.01). Conclusions PNI can be easily calculated, and may be used as a relatively new prognostic indicator for advanced colorectal cancer in clinical practice.

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, safety and advantages of gasless unilateral axillary approach (GUAA) in endoscopic thyroid surgery.Methods:A total of 334 patients who underwent the GUAA endoscopic thyroid surgery (GUAA group) or conventional open thyroid surgery (OS group) in the Department of Head and Neck Surgery of Zhejiang Cancer Hospital from January 2017 to June 2018 were retrospectively analyzed. There were 45 males and 289 females, aged from 12 to 72 years old, of whom 139 patients were assigned to GUAA group and 195 patients to OS group. Pathological results included papillary thyroid carcinoma (282 cases), nodular goiter (41 cases) and thyroid adenoma (11 cases). Surgical exploration development curve of GUAA group was drawn and was divided into two parts: the technical exploration stage and the technical stable stage. Surgical efficiency, incidences of complications, and incision satisfaction were compared between GUAA group in technical stable stage and OS group. SPSS 25.0 software was adopted for statistical analysis.Results:The mean age in GUAA group was younger than that in OS group, with a significant difference [(35.3±9.5) years vs. (48.1±10.6) years, t=11.31, P<0.01]. The cases in the endoscope group were divided into technical exploration stage for 51 cases and technical stable stage for 88 cases according to the exploration and development curve. In unilateral radical thyroidectomy and unilateral thyroid lobectomy, the mean operation time [(90.6±18.6) min and (93.5±22.0) min] and postoperative drainage volumes [(121.5±87.6) ml and (155.5±69.1) ml] of GUAA group in the stable stage were more than those of OS group [(61.6±15.6) min and (46.5±8.4) min] and [(93.2±42.3) ml and (78.9±48.7) ml]. The difference was statistically significant ( t=12.28, 7.23, 3.35 and 3.05 respectively, all P<0.05), but there were no significant differences in surgical bleeding volumes between two groups [(12.7±6.8) ml vs. (13.5±7.7) ml, t=0.74, P>0.05 and (16.3±14.1) ml vs. (11.9±5.1) ml, t=1.05, P>0.05]. Compared with OS group, GUAA group had the lower incidence of anterior cervical discomfort during swallowing (2.3% vs. 29.2%, P<0.01) and the higher incision satisfaction score (1.1±0.5 vs. 2.8±0.7, t=21.12, P<0.01), however, GUAA group had the higher incidence of supraclavicular (or infraclavicular) numbness after surgery (5.7% vs. 0, P<0.01). And there was no significant difference in the incidences of temporary recurrent laryngeal nerve injury, bleeding, hematoma, infection, lymphatic leakage or chylous leakage after surgery between two groups ( P>0.05). Conclusion:GUAA endoscopic thyroid surgery is a safe method with high cosmetic satisfaction.

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*