The efficient clinical treatment of oral squamous cell carcinoma(OSCC)is still a challenge that demands the development of effective new drugs.Phenformin has been shown to produce more potent anti-tumor activities than metformin on different tumors,however,not much is known about the influence of phenformin on OSCC cells.We found that phenformin suppresses OSCC cell proliferation,and promotes OSCC cell autophagy and apoptosis to significantly inhibit OSCC cell growth both in vivo and in vitro.RNA-seq analysis revealed that autophagy pathways were the main targets of phenformin and identified two new targets DDIT4(DNA damage inducible transcript 4)and NIBAN1(niban apoptosis regulator 1).We found that phenformin significantly induces the expression of both DDIT4 and NIBAN1 to promote OSCC autophagy.Further,the enhanced expression of DDIT4 and NIBAN1 elicited by phenformin was not blocked by the knockdown of AMPK but was suppressed by the knockdown of transcription factor ATF4(activation transcription factor 4),which was induced by phenformin treatment in OSCC cells.Mechanistically,these results revealed that phenformin triggers endoplasmic reticulum(ER)stress to activate PERK(protein kinase R-like ER kinase),which phosphorylates the transitional initial factor eIF2,and the increased phosphorylation of eIF2 leads to the increased translation of ATF4.In summary,we discovered that phenformin induces its new targets DDIT4 and especially NIBAN1 to promote autophagic and apoptotic cell death to suppress OSCC cell growth.Our study supports the potential clinical utility of phenformin for OSCC treatment in the future.

Objective:To evaluate clincal value of preoperative peripheral blood CD4/CD8 and neutrophil to lymphocyte ratio (NLR) in papillary thyroid carcinoma (PTC) coexisted with Hashimoto’s thyroiditis (HT) .Methods:Clinicopathological data of 202 patients diagnosed as PTC treated with operation from Jul.2016 to Jun.2019 were retrospectively analyzed. They were divided into Treatment Group including 94 PTC coexisted with HT and Control Group including 108 thyroid cancer according to the postoperateive pathology report. CD4+ and CD8+ subsets in peripheral blood were analyzed by flowcytometer and blood counts were measured before surgery.Results:There was no significant difference in gender, tumor size, number of lesions or lymph node metastasis between the two goups. In comparison with Control Group, median age was lower (39.5 vs 50.5, P=0.001) and CD4/CD8 raito (1.9731.973 Cvs 1.24141973 CD P=0.001) was higher in Treatment group. There was a higher proportion of bilateral lobe thyroidectomy in Treatment Group (40/94 vs 26/108, P=0.005) . A multivariate model analysis identified CD4/CD8 raito as independent risk factor for incidence of PTC coexisted with HT [ OR=0.035, 95% CI (0.009-0.093) , P=0.001]. The NLR level of thyroid cancer patients was correlated with lateral lymph node metastasis negatively (correlation coefficients=-0.286, P=0.045) . Conclusions:PTC might have some connection with HT mediated by body inmune status. Preoperative NLR level is correlated with lateral lymph node metastasis.

Objective To explore the characteristics of immunological changes in tree shrews infected with orthoreovirus, and provide a theoretical basis for the prevention of virus in tree shrews. Methods 40 -50-day-old tree shrews were divided into three groups: MRV1/TS/2011 virus-infected and MRV3/TS/2013 virus-infected groups, and saline-treated control group. On the 1, 8, 14, 21, and 28 days after infection, blood samples were taken from the tail vein and used for RT-PCR, flow cytometry and ELISA detection, to assess the viral load, number of CD4/CD8/CD19 cells, and IFN-gamma expression. Results The MRV1/TS/2011 and MRV3/TS/2013 viral load in the plasma and the number of CD4 +and CD19 +cells reached a peak at the 14th day after infection. At the first day after MRV1/TS/2011 infection, the CD4 +cells had a significantly higher expression compared with the normal group. CD8 +cells and the IFN-gamma expression reached a peak at the 21st day after infection. The expression of CD4 +was even higher after MRV1/TS/2011 infection, and the expression of CD8 +cells was higher after MRV3/TS/2013 infection. Conclusions We would conclude that after MRV1/TS/2011 and MRV3/TS/2011 virus infection, accompanying the changes of viral load, it shows some regularity of the expression of CD4/CD8/CD19 and IFN-gamma in the tree shrews: at the early stage of MRV1/TS/2011 virus infection, humoral immunity is stimulated, and CD4 +cells play a major role. MRV3/TS/2013 virus may mainly affect the cellular immunity, while humoral immunity only plays a role at a high viral expression or the late stage of infection. CD4 +cells may be more sensitive to type 1 reovirus, and CD19 +cells may be more sensitive to type 3 reovirus.

Objective To investigate degrees of fibrosis of adenomyosis(AM)myometrium and explore its relationship with dysmenorrhea. Methods Thirty AM patients who had hysterectomy from July, 2015 to December, 2016 in Beijing Obstetrics and Gynecology Hospital were selected as AM group; 28 cases of hysterectomy due to cervical lesions(none AM)were selected as control group. The area ratio of collagen fiber in the two groups was analysed by modified Masson stain, and the expression of collagen type Ⅰprotein in the two groups was analysed by immunohistochemical method. Results (1)The degree of fibrosis:the area ratio of collagen fiber and the expression of collagen type Ⅰof AM group [(34.5±5.1)％, 0.23±0.06] were significantly higher than those of control group [(26.7±10.1)％,0.18±0.08; all P<0.05].(2)The relationship between the degree of fibrosis and dysmenorrhea: the area ratio of collagen fiber and the expression of collagen type Ⅰ in severe dysmenorrhea, moderate dysmenorrhea, and none-mild dysmenorrhea were(35.3± 4.3)％,0.25±0.05;(35.7±3.2)％, 0.26±0.06;(25.0±2.9)％,0.15±0.03, there were significantly different among them(all P<0.01). And the area ratio of collagen fiber, the expression of collagen type Ⅰ were positively correlated with the degree of dysmenorrhea(r=0.50, 0.50; all P<0.05). Conclusions The area ratio of collagen fiber and the expression of collagen type Ⅰin AM are higher than in control group, and positively correlated with the severity of dysmenorrhea. These results suggest the degrees of fibrosis might be correlated with dysmenorrhea.

Objective To investigate the antitumor activity of the procaspase-3 activator SM-1 in BGC-823 cells in vivo and in vitro and the mechanisms.Methods The inhibitory effects of SM-1 on proliferation of BGC-823 cells were evaluated using MTT method, the cell apoptosis rate was detected by flow cytometry, and the expression of caspase-3 protein and procaspase-3 mRNA was detected by Western blotting and RT-PCR, respectively.SM-1 Antitumor activity was evaluated using the xenograft of BGC-823 cells in nude mice.Results SM-1 effectively inhibited the proliferation in vitro and in-duced apoptosis of BGC-823 cells in a dose-dependent manner.After treatment with SM-1 for 48 h, the protein expression levels of caspase-3 and mRNA expression levels of procaspase-3 were increased.SM-1 significantly inhibited growth of BGC-823 xenograft tumor at the 300 mg/kg dose and the inhibition rate was 56.3%(P<0.05).Conclusion SM-1 can significantly inhibit the tumor growth of BGC-823 cells in vivo and in vitro.The mechanism is possibly related to the activation of procaspase-3 and induced apoptosis of tumor cells.

Objective To investigate the diagnostic value of cyclophilin A(CyPA)for chronic heart failure (CHF).Methods Eighty pateints,made a definite diagnosis as CHF,were classified 30 cases as A phase,29 cases as C phase,21 cases as D phase of them.30 healthy subjects were enrolled in this study.Serum CyP A was detcted by ELISA.NT-proBNP was detec-ted with Roche Cobas 6 000 automatic electrochemilu-minescence immunoassay system.Results The amounts of serum CyP A(x-±s,ng/ml)in healthy subjects and each group with CHF were 110.10±49.73,327.85±82.67,331.70±69.34 and 342.46±92.55.Using Oneway Anova,CyP A concentration of CHF was significantly higher than healthy controls (F=58.45,P<0.01).Further using Post Hoc Tests-Multiple Comparisons,the CyP A concentration of each group with CHF was significantly higher than the healthy control group (Mean differences were 217.75~232.36,P<0.01),but showed no significant difference between the groups with CHFs (Mean differences were 3.37~14.61,P>0.05).Results of ROC curves showed the AUC (CyP A)was 0.97.The best cutoff value was 198.39 (ng/ml).The sensitivity was 91.30%,the specificity was 93.30%.The correlation analysis showed that CyP A and NT-proBNP were correlated (r=0.30,P<0.01). CyP A and NT-proBNP combined detections showed the sensitivity was 93.80% and the specificity was 100%.Conclusion The Cyp A levels in CHF group are significantly higher than the control group,suggesting that Cyp A may be a factor for CHF appearance.These results indicate that combined detections may helpful to early diagnosis of CHF.

Objective To investigate the significance of Ang-Ⅱ in diagnosis of chronic heart failure.Methods Eighty patients with chronic heart failure were divided into 3 groups:A phase with 30 cases,as C phase with 29 cases and D phase with 21 cases.30 healthy subjects were enrolled in this study.Serum Ang-Ⅱ was detected by ELISA.NT-proBNP was detected with Roche Co-bas6000 automatic electrochemiluminescence immunoassay system.Results Using Oneway Anova,the Ang-Ⅱconcentration of pa-tients with chronic heart failure was significantly higher than that of the healthy subjects (F =38.35,P <0.01).Further using Post Hoc Tests-Multiple Comparisons,the Ang-Ⅱ concentration of each group with chronic heart failure was significantly higher than that of the healthy control group.Results of ROC curves showed the aera under the curve of Ang-Ⅱwas 0.92.The best cutoff value was 552.25 (ng/mL).The sensitivity was 90.00%,and the specificity was93.30%.The correlation analysis showed that Ang-Ⅱwere correlated with NT-proBNP(r=0.23,P <0.05).The sensitivity and the specificity of combined detection of Ang-Ⅱand NT-proBNP were 98.80% and 100%,respectively.Conclusion The Ang-Ⅱlevels in chronic heart failure groups are significantly high-er than that in the control group,which indicates that combined detection may be helpful to the early diagnosis of chronic heart fail-ure.

Objective To establish an osteoporosis model in tree shrews by bilateral ovariectomy.Methods One hundred healthy 6-24 month old female tree shrews were used in this study.The diagnosis and treatment of osteoporo-sis in the model tree shrews was assessed according to the Interpretation of the Clinical Practice Guideines for Primary Oste-oporosis 2011 released by the Chinese Medical Association of Osteoporosis and Bone Mineral Salt Disease Branch.① Ac-cording to the human natural life span, sexual maturation time and peak bone mineral density ( PBMD) and the tree shrew natural life span and sexual maturation time, we estimated the PBMD of tree shrew in the age of months.Six 6-, 12-, 18-and 24-month old tree shrews in each group were randomly taken for measurement of BMD, and compared the differences between groups to verify the PBMD age of tree shrews.②We chose 62 6-month old tree shrews from the rest of tree shrews to measure their BMD to determine the mean and standard deviation of BMD.Then we divided them into study group ( n =32) and control group (n =30).Bilateral ovariectomy was performed in the study group, while omentectomy was done in the control group.BMD was measured at 3, 6, 9, 12 months after the surgery, 6 animals from each group.The animals which met the criteria of guideline were taken for pathological examination.Results ①The PBMD of 6-, 12-, 18-, 24-month old tree shrews was 0.138 ±0.012, 0.143 ±0.010, 0.141 ±0.009, and 0.139 ±0.009) g/cm2 , respectively, showing no significant differences among them (P>0.05).②The mean PBMD of the tree shrews was 0.143 ±0.016 g/cm2 .③In the model groups, the number of tree shrews whose BMD met the criteria of guideline was 0, 5, 4, 5, respec-tively.The pathological examination of the proximal tibia from the model group showed sparse and some disrupted trabecu-lae and enlarged inter-trabecular space.In the control group, the tibia tissues showed normal histology of bone structure and no one tree shrew showed a BMD value which met the criteria of diagnosis of osteoporosis.Conclusions An osteopo-rosis model in tree shrews can be established at 6 months after bilateral ovariectomy, showing similar manifestation of osteo-porosis in postmenopausal women.The success rate of this modeling method is high and the model is stable.

Objective To establish a simple and useful kidney or bladder orthotopic tumor model used in preclinical pharmacodynamic evaluation.Methods Mouse model of orthotopic renal cancer were established by subrenal capsule implantation.After aspirating urine and irrigating bladder with PBS,the bladder urothelium was slightly impaired to establish the orthotopic bladder tumor model.Then, B-Ultrasound and H&E staining were used to confirm the availability.Results Tumors could be seen 2 weeks after surgery, accompanied by body mass loss of the mice.H&E staining showed that the tumor cells acted as infiltrative growth.The growth of tumor was inhibited by NTX in vivo, the tumor mass inhibitory rate of the KCC-853 orthotopic tumor model was 57.5% of 60 mg/kg NTX treatment and 48.8% in the T24 orthotopic tumor model of 30 mg/kg NTX treatment.Conclusion Our methods for establishing the orthotopic kidney or bladder tumor model are simple and practical.The results indicate that nitroxoline has potential antitumor activity.

Objective To understand the histological characteristics of the major endocrine organs of tree shrew , and provide a normal histological atlas of endocrine organs of tree shrew .Methods Ten artificially fed healthy tree shrews were killed and dissected after anesthesia .The thyroid, parathyroid, adrenal and pituitary glands were observed by gross inspection and samples were taken for routine histological examination with HE staining .Results ( 1 ) The thyroid gland was pale yellow, located on both sides of the 2-4 tracheal rings.The thyroid gland was plate-shaped, its surface was covered with a thin fibrous capsule . The thyroid parenchyma was divided into several lobules by stretched capsule membrane .Follicular and parafollicular cells were distributed in the lobules , and red colloid was present in follicular cavity.(2) Each side had one parathyroid , located on the cranial or the outer surface of the middle part of the thyroid gland, and was slightly covered by thyroid .The gland was round or oval , and its parenchyma was made up of the principal cells and eosinophil cells , and acinar structure appeared in the parenchyma .( 3 ) The adrenal glands were oval , yellow color, located in the renal hili , and linked to the kidneys .They were surrounded by a thin capsule .The parenchyma was divided into cortex and medulla .The cortex was divided into zona glomerulosa , zona fasciculata and zona reticularis from outside to inside.The zona glomerulosa was the thickest layer and the zona fasciculata was the thinnest .The medulla cells formed clumps or mesh, with central vein in the central part .(4) The pituitary gland was located in the sella turcica , with no recessus hypophysis .The pituitary gland was composed of the adenohypophysis and neurohypophysis .Its surface was covered with a connective tissue capsule .The pituitary gland was divided into distal part , middle part and pars tuberalis . neurohypophysis was made up of neural and pars infundibularis .Conclusions The histological atlas of endocrine organs in the tree shrew is established , which is close to that of the primate animals in the morphology , and provide histological evidence for the study of tree shrew endocrine organs and disorders , as well as the animal model of human diseases .