BACKGROUND:Overactive osteoclasts disrupt bone homeostasis and play a bad role in the pathological mechanisms of related skeletal diseases,such as osteoporosis,fragility fractures,and osteoarthritis.Studies have confirmed that ellagic acid and ellagtannin have the potential to inhibit osteoclast differentiation.As their natural metabolites,urolithin A has antioxidant,anti-inflammatory,anti-proliferative and anti-cancer effects,but its effect on osteoclast differentiation and its underlying molecular mechanisms remain unclear. OBJECTIVE:To explore the effect of urolithin A on osteoclast differentiation induced by receptor activator for nuclear factor-κB ligand and its mechanism. METHODS:Mouse mononuclear macrophage leukemia cells(RAW264.7)that grew stably were cultured in vitro.Toxicity of urolithin A(0,0.1,0.5,1.5,2.5 μmol/L)to RAW264.7 cells were detected by cytotoxic MTS assay to screen out the safe concentration.Different concentrations of urolithin A were used again to intervene with receptor activator for nuclear factor-κB ligand-induced differentiation of RAW264.7 cells in vitro.Then,tartrate-resistant acid phosphatase staining and F-actin ring and nucleus staining were performed to observe its effect on the formation and function of osteoclasts.Finally,the expressions of urolithin A on upstream and downstream genes and proteins in the MAPK signaling pathway were observed by western blot and RT-qPCR assays. RESULTS AND CONCLUSION:Urolithin A inhibited osteoclast differentiation and F-actin ring formation in a concentration-dependent manner and 2.5 μmol/L had the strongest inhibitory effect.Urolithin A inhibited the mRNA expression of Nfatc1,Ctsk,Mmp9 and Atp6v0d2 and the protein synthesis of Nfatc1 and Ctsk,related to osteoclast formation and bone resorption.Urolithin A inhibited the activity of osteoclasts by downregulating the phosphorylation of p38 protein to inhibit the mitogen-activated protein kinase signaling pathway.

Objective To investigate the clinical effects and pregnancy outcomes of using luteal phase long protocol and GnRH antagonist protocol in patients with polycystic ovary syndrome(PCOS)who have failed their first GnRH antagonist protocol therapy.Methods The clinical data of 163 PCOS patients who underwent IVF/ICSI-ET were retrieved.After the failure of their first GnRH antagonist protocol treatment,they were divided into two groups in the second controlled ovarian hyperstimulation(COH)cycle:Luteal phase long protocol group(n=95)and Gn-RH antagonist protocol group(n=68).A retrospective analysis and comparison of basic clinical data,clinical and laboratory indicators,and pregnancy outcomes between two groups were conducted.Results ① There was no sta-tistically significant difference in basic clinical indicators between two group except LH.② Compared the first and second cycle treatments of patients in the luteal phase long protocol group,the initiation dose of gonadotropin(Gn),total number of Gn days,total Gn usage,estradiol(E2)on the day of hCG injection,number of retrieved eggs,oocyte maturation rate,2PN fertilization rate,2PN cleavage rate,blastocyst formation rate,high-quality blas-tocyst formation rate,and moderate to severe OHSS rate were significantly higher than those in the first GnRH an-tagonist cycle(P＜0.05).The GnRH antagonist protocol group also showed similar improvements.③ The com-parison of the second COH cycle between two groups showed that the total number of Gn days,total Gn usage,and total Gn cost in the luteal phase long protocol group were significantly higher(P＜0.05),while the E2 and LH on the day of hCG injection,and the maturation rate of eggs were significantly lower than those in the GnRH antagonist protocol group(P＜0.05).However,there was no statistically significant difference in the number of retrieved eggs,2PN fertilization,2PN cleavage,blastocyst formation rate,high-quality blastocyst formation rate,and OHSS rate between the two groups;④ The comparison of fresh transplantation cycles for the second COH cycle between the two groups showed that the luteal phase long protocol fresh transplantation rate,implantation rate,clinical preg-nancy rate,and live birth rate were slightly higher than those of the GnRH antagonist protocol group,but the differ-ence was not statistically significant.Comparing the outcomes of pregnancy following the initial frozen-thawed em-bryo transfer(FET)between two groups,the biochemical pregnancy rate and clinical pregnancy rate of the GnRH antagonist protocol group were higher than those of the luteal phase long protocol group(P＜0.05).However,no significant statistical variations were found in implantation rate,live birth rate,neonatal gestational age,and birth weight.Conclusion For PCOS patients who fail the first GnRH antagonist protocol,an appropriate increase in the initiating dose and usage of Gn can achieve satisfactory pregnancy outcomes with both protocols.Compared with change to a luteal phase long protocol,reusing the GnRH antagonist protocol still maintains its long-standing advan-tages,such as shorter total Gn days,lower costs,and better patient compliance.

COVID-19 is caused by a novel coronavirus-severe acute respiratory syndrome coronavirus type 2 (SARS-CoV-2), which has being spreading around the world, posing a serious threat to human health and lives. Neutralizing antibodies and small molecule inhibitors for virus replication cycle are the main antiviral treatment for novel coronavirus recommended in China. To further promote the rational use of antiviral therapy in clinical practice, the National Center for Infectious Diseases (Beijing Ditan Hospital Capital Medical University and the First Affiliated Hospital, Zhejiang University School of Medicine) invited experts in fields of infectious diseases, respiratory and intensive care to develop an Expert Consensus on Antiviral Therapy of COVID-19 based on the Diagnosis and Treatment Guideline for COVID-19 ( trial version 10) and experiences in the diagnosis and treatment of COVID-19 in China. The consensus is concise, practical and highly operable, hopefully it would improve the understanding of antiviral therapy for clinicians and provide suggestions for standardized medication in treatment of COVID-19.

Objective:To investigate the effect and molecular mechanism of tumor necrosis factor-α-inducible protein 8-like 2(TIPE2)on lipopolysaccharide(LPS)or interleukin-4(IL-4)-induced phenotypic switch of adipose tissue macrophages(ATM).Methods:The expression levels of TIPE2, inducible nitric oxide synthase(iNOS), monocyte chemoattractant protein 1(MCP-1), CD206, and arginase 1(Arg-1)in the visceral adipose tissue of obese mice, TIPE2-knockout(KO)mice, and control mice were detected by immunohistochemistry, Western blotting, and real-time PCR(RT-qPCR). Peritoneal macrophages isolated from KO and wild-type mice and RAW 264.7 mouse macrophage cell line were cultured, and then stimulated with LPS(100 ng/mL)or IL-4(20 ng/mL)for 6 hours. The expression levels of TIPE2, iNOS, MCP-1, CD206, and Arg-1 were detected by Western blotting and RT-qPCR.Results:Obese mice showed down-regulated TIPE2 expression, up-regulated pro-inflammatory markers iNOS and MCP-1 expressions, and down-regulated anti-inflammatory markers CD206 and Arg-1 expressions. LPS decreased the expression of TIPE2 in RAW 264.7 cells and peritoneal macrophages from mice, increased the expression of the classically activated macrophages(M1 phenotype)markers iNOS and MCP-1, and decreased the expression of the substituting activated macrophages(M2 phenotype)markers CD206 and Arg-1. IL-4 increased the expression of TIPE2 in RAW 264.7 cells and peritoneal macrophages, decreased the expression of iNOS and MCP-1, and increased the expression of CD206 and Arg-1. During the M1 polarization of macrophages, LPS increased toll-like receptor(TLR4)expression as well as nuclear transcription factor κBα suppressor protein(IκBα) and NF-κB phosphorylations in macrophages. Knockout of TIPE2 further increased the expression of the TLR4/IκBα/NF-κB signaling pathway and M1 macrophage markers, and further reduced the expression of the M2 macrophage markers.Conclusion:TIPE2 regulates ATM phenotypic transformation through inhibition of the TLR4/IκBα/NF-κB signaling pathway, which ameliorates adipose tissue inflammation in obese states.

Objective 　To evaluate the role of surgical resection on synchronous multiple pulmonary nodules identified difficultly in clinics. Methods 　The clinical data of 97 patients with synchronous multiple pulmonary nodules who received surgical resection between 2012 and 2019 in Hunan Cancer Hospital were retrospectively analyzed. There were 72 males and 25 females, aged 58.1卤9.0 years. Among these patients, there were 78 patients with ipsilateral and 19 patients with bilateral pulmonary nodules. Clinicopathological parameters between main nodules and secondary nodules were evaluated. Perioperative morbidity was also assessed. Results 　The operation was successfully completed on all patients for the ipsilateral and bilateral lesions. Totally, 71.1% of mian lesions was mostly removed by lobectomy, and the completion rate of video-assisted thoracoscopic surgery (VATS) was 69.1% (67/97); 80.4% of secondary lesions were mostly removed by wedge resection, and the completion rate of VATS was 71.1% (69/97). The incidence of grade 3 or higher complications after unilateral or bilateral surgery was 12.8% and 5.3%, respectively. Postoperative pathology confirmed that the main lesions were malignant in 65 patients (67.0%), mainly adenocarcinoma (63.1%), of which 43.1%were in the stage Ⅰ; 32 patients were benign, mainly tuberculoma (56.3%). There were 29 patients of malignant secondary lesions, 67 benign, and 1 both benign and malignant; the pathological agreement rate of primary and secondary lesions was 54.6% (lung cancer metastases in the lung and all the benign). When the primary lesion was malignant with its diameters of <3 cm, 3-<5 cm, 5-7 cm, >7 cm, the metastatic rate of secondary lesions was 42.5%, 15.8%, 20.0%, 0, respectively. When the primary lesion was malignant with lymph node metastasis, the probability of the secondary lesion being a metastatic tumor was higher than that without lymph node metastasis (46.7% vs. 30.0%, P>0.05). When the primary lesion was malignant and the primary and secondary lesions were located in the same lobe, the secondary lesions were more likely to metastasize (54.5%), while when they were located on different lobes on the same side or different sides, the secondary lesions were more likely to be benign (58.1%, 72.7%), and the possibility of metastasis was small ( 32.6%, 9.1%). When the primary lesion was benign and clinical differential diagnosis was difficult, the secondary lesion was benign. Conclusion    For synchronous multiple pulmonary nodules, the diameter of the primary lesion is large, the metastatic rate of secondary lesions tends to decrease. In ipsilateral synchronous multiple pulmonary nodules, especially with node metastasis, the risk of metastatic nodule increases. Bilateral surgical resection does not significantly increase the perioperative morbidity.

Objective:To investigate the stiffness characteristic of triple-negative human breast cancer at different size in a implantation nude female mice model using shear wave elastography(SWE) and to compare the clinical pathologic features of tumors with elasticity variables.Methods:Human breast cancer MDA-MB-231 cells were injected into 30 nude female mice and 27 transplanted tumors were successfully found in nude female mice. Ultrasound and SWE were longitudinally performed on maximum diameter plane of 21 tumours in 21 nude mice. The elastic parameters of maximal elasticity(Emax), mean elasticity (Emean) and standard deviation of elasticity(Esd) were recorded. The mice were divided into 3 groups according to the tumor size. They were group A with tumor size less than or equal to 5 mm, group B with tumor size greater than 5 mm and less than or equal to 10 mm, group C with tumor size larger than 10 mm and smaller than or equal to 15 mm. Compared with pathology, the relationships between Ki67 of transplanted tumor and elastic parameters were analyzed.Results:As the transplanted tumors increased, the values of Emax, Esd, Ki67 all increased. The lesions maximal size, Emax, Esd, Ki67 were significant higher in group B ( P<0.001, P=0.006, P=0.002, P=0.026) and group C ( P<0.001, P<0.001, P<0.001, P=0.028) than group A. The other parameters were not significantly different among the groups(all P>0.05). The size of transplanted tumors was significantly and positively correlated with Emax ( rs=0.673, P=0.001), Esd ( rs=0.661, P=0.001), and Ki67 ( rs=0.509, P=0.018). Conclusions:SWE Emax and Esd can reflect the tumor tissue stiffness change and biological activity during the tumor growth.

Objective: The aim of this study is to evaluate the commutability of 16 processed materials for 17-hydroxyprogesterone by using 2 commutability assessment approaches. Methods: 52 serum specimens were collected in Clinical Laboratory Department of Beijing Hospital from February 2018 to June 2019. According to the report of the Clinical and Laboratory Standards Institute (EP14-A3) document and the recommendations of the International Federation of Clinical Chemistry and Laboratory Medicine (IFCC) working group on commutabilityassessment, serum 17-hydroxyprogesterone isotope diluent chromatogram tandem mass spectrometry (ID-LC/MS/MS) was used for comparison. Three clinical routine analysis systems (1 radioimmunoassay, 2 LC/MS analysis methods) were used to determine the concentration of 17-hydroxyprogesterone in 52 human serum samples and 16 processed materialsfor commutabilityassessment. Results: Combined with the results of the two commutability assessment, all accuracy verification materials and national steroid hormone standards showed good commutability in the LC/MS analysis system, and 6/9 EQA materials showed commutability in the three routine analysis systems.All materials showed good commutability in the LC/MS analysis system of bias difference method. Conclusions The two kinds of commutability assessment results are different. Bias difference method has more clinical value, but it has certain application limitations. The use of fresh frozen human serum as a quality assessment materialfor serum 17-hydroxyprogesterone is meets the commutability requirement.

Aldosterone and renin are important clinical indicators in the diagnosis and treatment of hypertension. There are two kinds of methods for aldosterone detection, including immunoassay and mass spectrometry. The coefficients of variation within the methods, as well as the deviation between methods are large.And the results of immunoassay are much higher than that of mass spectrometry. Clinically, renin tests mainly include renin concentration and renin activity, both of which have advantages and disadvantages in the diagnosis and treatment of hypertension. The disordered test results of aldosterone and renin brings great trouble to the clinical diagnosis and treatment. Standardization/consistency is urgently needed to make the test results of each laboratory more accurate and comparable. The standardization project for renin and aldosterone in China will greatly promote the standardization process in order to provide accurate and comparable clinical tests.

Objective To evaluate the risk factors in terms of clinical characteristics ,serological indicators and sonographic features regarding thyroid papillary carcinoma ( PTC) for the central neck lymph node metastasis . Methods One thousand one hundred and seventy‐four patients accepted thyroid cancer surgery at Fudan University Shanghai Cancer Center from January to September 2016 were enrolled . All patients were confirmed to be solitary PTC in paraffin wax pathology after thyroidectomy .M eanwhile ,the status of central neck lymph node metastasis was determined referring to postoperative pathology . All features of the PTC lesion in terms of clinical ,serological and sonographic features were evaluated for the association with central neck lymph node metastasis using univariate and multivariate logistic regression analysis . Meanwhile ,a nomogram model was established for the determined risk factors . Results Out of 1 174 patients ,469 patients ( 39 .9% ) presented central neck lymph node metastasis ,univariate analysis showed that age , gender , preoperative thyroglobulin ( Tg ) and thyroid peroxidase antibody ( T POAb ) , maximum diameter ,location ,close to the thyroid capsule ,AP/T R ,echo ,acoustic halo ,and presence of microcalcification were related with CLNM ( P ＜ 0 .05 ) . M ultivariate logistic regression analysis demonstrated that less than 55 years‐old , male , Tg higher than 20 eg/L , T POAb less than 1 kU/L , maximum diameter larger than 10 mm ,and presence of microcalcification were independent risk factors for CLNM . T he nomogram was established based on independent risk factors determined by the logistic regression with the AUC 0 .714 ,specificity 73 .1% ,and sensitivity 59 .7% . Conclusions For patients with single focal PTC lesion , younger age , male , higher Tg , lower T POAb , and larger lesions containing microcalcificatin on ultrasound are associated with central neck lymph node metastasis .

Objective: To evaluate the risk factors in terms of clinical characteristics and sonographic features regarding solitairy thyroid papillary carcinoma (PTC) for the lateral cervical lymph node metastasis(LLNM) and then to establish nomogram model. Methods: All patients were confirmed to be solitary PTC in paraffin wax pathology after thyroidectomy in Fudan University Shanghai Cancer Center from January to September 2016. Meanwhile, the status of lateral cervical lymph node metastasis was determined referring to postoperative pathology. Clinical characteristics including gender, age, preoperative thyroid stimulating hormone (TSH), thyroglobulin (Tg), thyroglobulin antibody (TGAb), thyroid peroxidase antibody (TPOAb), central lymph node metastasis (CLNM) and sonographic features of the PTC lesion including maximum tumor diameter, location, aspect ratio, relation with thyroid capsule, echo, margin, acoustic halo, microcalcification were evaluated for the association with lateral cervical lymph node metastasis using univariate and multivariate logistic regression analyses. Then the nomogram model was established and its application value was evaluated using ROC. Results: Out of 1 174 patients, 125 patients (10.6%) presented lateral neck lymph node metastasis and 10 patients presented skipping metastasis.Univariate analysis showed gender, preoperative Tg and TGAb, CLNM, maximum tumor diameter, location, close to the thyroid tumor capsule, echo, aspect ratio, acoustic halo, microcalcification were associated with LLNM(P<0.05). Binomial logistic regression analysis indicated CLNM, maximum tumor diameter of larger than 10 mm, superior or multiple location, microcalcification were independent risk factors of LLNM. The AUC of the nomogram model was 0.865, the sensitivity was 88.0%, the specificity was 75.2%, and the accuracy was 76.6%. Conclusions As for patients with single focal PTC, CLNM, larger lesions, microcalcification, superior location are associated with lateral neck lymph node metastasis. The nomogram model can be tried for clinical application.