Objective To study the expression of N6-methyladenosine(m6A)methyltransferase-like protein 16(METTL16),Ankyrin repeat and SOCS box containing protein 4(ASB13)in elderly breast cancer tissues and their relationship with clinicopathological features and prognosis.Methods 124 elderly patients with breast cancer diagnosed and treated in the Zibo 148 Hospital from January 2019 to January 2021 were selected.Immunohistochemistry(IHC)was used to detect the expression of METTL16 and ASB13 proteins in tissues.Real time fluorescence quantitative PCR(RT-qPCR)was used to detect the expression of METTL16 mRNA and ASB13 mRNA in tissues.Pearson correlation was used for correlation analysis.Survival analysis used Kaplan-Meier curves and Log Rank tests to compare the prognostic differences among patients with different METTL16 and ASB13 expression levels.COX regression analysis was used to analyze the prognostic factors of elderly patients with breast cancer.Results The positive rate of METTL16 in cancer tissue(70.97%)was higher than that in adjacent tissues(8.06%),while the positive rate of ASB13(22.58%)was lower than that in adjacent tissues(72.58%),the differences were statistically significant(χ2=102.642,62.146,all P<0.001).METTL16 mRNA,Snail mRNA and N-cadherin(N-cd)mRNA were higher in cancer tissues than in adjacent tissues,while ASB13 mRNA was lower in adjacent tissues,the differences were statistically significant(t=40.498～48.624,all P<0.001).METTL16 mRNA was positively correlated with Snail mRNA and N-cd mRNA in cancer tissue(r=0.712,0.669,all P<0.001),while ASB13 mRNA was negatively correlated with Snail mRNA and N-cd mRNA(r=-0.734,-0.759,all P<0.001).Compared with TNM stage Ⅰ-Ⅱ and patients with high to medium differentiation,patients with TNM stage Ⅲ and low differentiation had higher positive rate of METTL16 in cancer tissue and lower positive rate of ASB13,the differences were statistically significant(χ2=5.340～10.370,all P<0.05).The 3-year progression free survival rate(73.86%)of the METTL16 positive group was lower than that of the METTL16 negative group(88.89%),the 3-year progression free survival rate(92.86%)of the ASB13 positive group was higher than that of the ASB13 negative group(73.96%),and the differences were statistically significant(Log Rank χ2=4.483,4.882,all P<0.005).METTL16 positive,TNM stage Ⅲ,poor differentiation were risk factors affecting the prognosis of elderly breast cancer,and ASB13 positive was protective factors(Wald χ2=5.056～9.267,all P<0.001).Conclusion METTL16 expression is up-regulated and ASB13 expression is down-regulated in elderly breast cancer,both of which are related to the expression of invasion and metastasis genes,and are new prognostic markers for elderly breast cancer.

ObjectiveTo investigate the analgesic effect of Tuina at the "Weizhong （BL 40）" on neuropathic pain in a rat model of chronic constriction injury （CCI） of the sciatic nerve and its potential central spinal mechanisms. MethodsThirty-two Sprague-Dawley rats were randomly divided into four groups （8 rats in each group）， sham-operated group， model group， Tuina group， and blockade group. The CCI model was established in the model group， Tuina group， and the blockade group by ligating the sciatic nerve with catgut， while the sham-operated group underwent only sciatic nerve exposure without ligation. From postoperative day 4 to day 14， rats in the Tuina group and the blockade group received Tuina manipulation at the "Weizhong （BL 40）" using a dynamic pressure distribution measurement system （5 N pressure， 2 Hz frequency， 10 min per session， once daily）. The blockade group also received intraperitoneal injections of the microglial inhibitor minocycline （10 mg/kg） once daily. The sham-operated and the model group underwent the same handling and fixation as the Tuina group without actual Tuina. Mechanical withdrawal threshold （MWT） and paw withdrawal latency （PWL） were measured before surgery and on day 3， 7， 10， and 14 post-surgery. Transmission electron microscopy was used to evaluate sciatic nerve injury and repair， measuring axon diameter and total myelinated fiber diameter to calculate the g-ratio. Western Blotting was performed to detect the protein levels of ionized calcium-binding adapter molecule 1 （Iba-1）， CD206， CD68， interleukin-10 （IL-10）， and β-endorphin （β-EP） precursor pro-opiomelanocortin （POMC） in the ipsilateral spinal dorsal horn. ResultsCompared with the sham-operated group， the model group showed significantly reduced MWT and PWL on day 3， 7， 10， and 14 （P＜0.01）. Compared with the model group， the Tuina group and the blockade group showed increased MWT and PWL on day 10 and 14 （P＜0.05）. Compared with the Tuina group， the blockade group exhibited higher MWT on day 7， 10， and 14， and higher PWL on day 10 （P＜0.05）. Sciatic nerve pathological morphology revealed intact and well-structured myelin in the sham-operated group， while the model group exhibited myelin collapse， distortion， and myelin ovoid formation. The Tuina group displayed partially irregular myelin with occasional myelin collapse， whereas the blockade group exhibited partial myelin irregularities and phospholipid shedding. Compared with the sham-operated group， the model group showed a decreased g-ratio and increased levels of Iba-1 and CD68 in the spinal dorsal horn （P＜0.05 or P＜0.01）. Compared with the model group， the Tuina group and the blockade group exhibited an increased g-ratio and reduced Iba-1 and CD68 levels. Additionally， the Tuina group showed elevated levels of CD206， IL-10， and POMC， whereas the blockade group had decreased CD206 levels （P＜0.05）. ConclusionTuina at "Weizhong （BL 40）" alleviates neuropathic pain in CCI rats， potentially by regulating microglial activation in the spinal cord， inhibiting M1 polarization while promoting M2 polarization， and activating the IL-10/β-EP pathway to exert analgesic effects.

Objective To study the expression of N6-methyladenosine(m6A)methyltransferase-like protein 16(METTL16),Ankyrin repeat and SOCS box containing protein 4(ASB13)in elderly breast cancer tissues and their relationship with clinicopathological features and prognosis.Methods 124 elderly patients with breast cancer diagnosed and treated in the Zibo 148 Hospital from January 2019 to January 2021 were selected.Immunohistochemistry(IHC)was used to detect the expression of METTL16 and ASB13 proteins in tissues.Real time fluorescence quantitative PCR(RT-qPCR)was used to detect the expression of METTL16 mRNA and ASB13 mRNA in tissues.Pearson correlation was used for correlation analysis.Survival analysis used Kaplan-Meier curves and Log Rank tests to compare the prognostic differences among patients with different METTL16 and ASB13 expression levels.COX regression analysis was used to analyze the prognostic factors of elderly patients with breast cancer.Results The positive rate of METTL16 in cancer tissue(70.97%)was higher than that in adjacent tissues(8.06%),while the positive rate of ASB13(22.58%)was lower than that in adjacent tissues(72.58%),the differences were statistically significant(χ2=102.642,62.146,all P<0.001).METTL16 mRNA,Snail mRNA and N-cadherin(N-cd)mRNA were higher in cancer tissues than in adjacent tissues,while ASB13 mRNA was lower in adjacent tissues,the differences were statistically significant(t=40.498～48.624,all P<0.001).METTL16 mRNA was positively correlated with Snail mRNA and N-cd mRNA in cancer tissue(r=0.712,0.669,all P<0.001),while ASB13 mRNA was negatively correlated with Snail mRNA and N-cd mRNA(r=-0.734,-0.759,all P<0.001).Compared with TNM stage Ⅰ-Ⅱ and patients with high to medium differentiation,patients with TNM stage Ⅲ and low differentiation had higher positive rate of METTL16 in cancer tissue and lower positive rate of ASB13,the differences were statistically significant(χ2=5.340～10.370,all P<0.05).The 3-year progression free survival rate(73.86%)of the METTL16 positive group was lower than that of the METTL16 negative group(88.89%),the 3-year progression free survival rate(92.86%)of the ASB13 positive group was higher than that of the ASB13 negative group(73.96%),and the differences were statistically significant(Log Rank χ2=4.483,4.882,all P<0.005).METTL16 positive,TNM stage Ⅲ,poor differentiation were risk factors affecting the prognosis of elderly breast cancer,and ASB13 positive was protective factors(Wald χ2=5.056～9.267,all P<0.001).Conclusion METTL16 expression is up-regulated and ASB13 expression is down-regulated in elderly breast cancer,both of which are related to the expression of invasion and metastasis genes,and are new prognostic markers for elderly breast cancer.

Objective To detecting the expression of nicastrin,N1ICD and hes1 proteins in normal liver tissues and liver cancer tissues of C57BL/6 mice.Methods Twelve 6-week-old male C57BL/6 mice were ran-domly equally divided into the control group and model group.In the model group,the in situ liver cancer model was established with injection of Hepa1-6 cells into the liver,and the control group was treated with injection of the same amount of normal saline.Liver cancer was verified by HE staining.The expression of nicastrin,N1ICD and hes1 proteins in the normal and cancerous liver tissues was detected by IHC and Western blot.Results IHC results showed that nicastrin,N1ICD and hes1 proteins were localized in the hepatic sinusoidal endothelial cells in the normal hepatocytes,but not expressed in the hepatocytes.The model group showed higher expression of nicas-trin protein but lower expression of N1ICD and hes1 protein in the cancer cells compared with the control group.Conclusion NCSTN gene may play a carcinogenic role in mouse hepatocellular carcinoma,while notch1 and hes1 may play a carcinostasis role in the carcinoma.

Objective:To investigate clinical manifestations, pathological features and diagnosis of eczematoid clear cell acanthoma of the nipple/areola.Methods:The clinical manifestations, histopathological features, special staining results and immunohistochemical features of a case of eczemtoid clear cell acanthoma of the nipple/areola firstly reported in China were analyzed, and compared with those of similar cases in foreign literature.Results:The female patient presented with recurrent pruritic rashes on the left nipple and areola for over 2 years. Skin examination showed hypertrophic skin on the left nipple and areola, and scattered erythema, hypopigmented macules and hyperpigmented macules on the areola, which were covered with a few crusts and scales. Histopathological examination of the skin lesions showed focal epidermal crusts and scales, focal parakeratosis, extended and fused rete ridges, thickened spinous layer, focal spongiosis, clear cell clumps in the spinous cell layer, telangiectasia in the superficial dermis, with infiltration of a few eosinophils and neutrophils. Periodic acid-Schiff staining showed positive results, and immunohistochemical staining revealed positive reaction for epithelial membrane antigen. Topical treatment with triamcinolone acetonide and econazole cream was effective, and topical application of 3% boric acid solution could alleviate exudation. During the 6-month follow-up, the patient experienced intermittent recurrence twice, and responded well to the above treatment.Conclusions:Eczematoid clear cell acanthoma of the nipple/areola has unique clinical and pathological features, revealing that it′s a new subtype of clear cell acanthoma. Pathological examination is the gold standard for its diagnosis.

Objective:To evaluate the predictive value of systemic immune inflammation index (SII) for short-term prognosis in patients with sepsis complicated with acute kidney injury who received continuous renal replacement therapy (CRRT) .Methods:From Feb. 2018 to Jan. 2022, 90 patients with sepsis complicated with acute kidney injury who received CRRT in Hangzhou Cancer Hospital were included as the research objects. According to the survival of patients within 90 days, they were grouped into a survival group of 60 cases and a death group of 30 cases. The clinical baseline data such as age, sequential organ failure assessment (SOFA) score, acute physiological function and chronic health status scoring system II (APACHE II), albumin (ALB) were recorded. Automatic blood cell analyzer was applied to detect lymphocyte count, platelet count and neutrophil count in peripheral venous blood of patients with sepsis complicated with acute kidney injury who received CRRT, and the SII was calculated; Pearson correlation analysis was performed to analyze the correlation between SII and clinical baseline data in dead patients with sepsis complicated with acute kidney injury receiving CRRT; ROC curve was drawn to analyze and compare the predictive value of SII, SOFA score and APACHEII for death in patients with sepsis complicated with acute kidney injury receiving CRRT; Kaplan-Meier curve was performed to describe the changes in patient survival; multivariate Cox regression analysis was performed to analyze the factors affecting 90-day death in patients with sepsis complicated with acute kidney injury receiving CRRT.Results:SII (2636.74), C-reactive protein (CRP) [ (92.45±29.74) mg/L], SOFA score (12.83±3.24) and APACHEII (28.30±7.51) in the death group were higher than those in the survival group [341.92, (50.24±16.13) mg/L, 10.00±3.12, 25.13±6.87], and ALB [ (2.50±0.53) g/dl] was lower than that in the survival group [ (2.79±0.61) g/dl] ( P<0.05). SII was significantly positively correlated with CRP, SOFA score and APACHEII, and significantly negatively correlated with ALB in patients with sepsis complicated with acute kidney injury who received CRRT ( P<0.05). The areas under the curve (AUC) of SII, SOFA score, and APACHEII for predicting death in patients with sepsis complicated with acute kidney injury who received CRRT were 0.936, 0.827, and 0.736, respectively, and AUC of SII prediction was greater than that of SOFA score and APACHEII ( P<0.05). The 90-day survival rate of patients with sepsis complicated with acute kidney injury who received CRRT in the high SII group (23/46, 50.00%) was lower than that in the low SII group (37/44, 84.09%) ( P<0.05). SII was an independent risk factor for death within 90 days in patients with sepsis complicated with acute kidney injury who received CRRT ( P<0.05) . Conclusion:SII can better predict the 90-day mortality risk of patients with sepsis complicated with acute kidney injury receiving CRRT, and assist clinical assessment of short-term prognosis.

Objective:To investigate the association between serum sex hormone-binding globulin (SHBG) and non-alcoholic steatohepatitis (NASH).Methods:In this cross-sectional study, a total of 371 middle-aged and young obese patients who were hospitalized and underwent liver puncture in Nanjing Drum Tower Hospital from January 2016 to April 2021 were included. The population was divided into control group ( n=43) and non-alcoholic fatty liver disease (NAFLD) group ( n=328) based on the non-alcoholic fatty liver disease activity score. Subjects in NAFLD group were further divided into non-alcoholic fatty liver (NAFL) ( n=60), uncertain-NASH ( n=172), and NASH ( n=96). Serum SHBG was tested in patients with NAFLD who were divided into three subgroups according to tertiles. The liver pathological characteristics in different SHBG level subgroups were compared. The risk factors of NASH were analyzed by logistic regression. The prediction model of NASH noninvasive diagnosis was established by forward stepwise regression, and the diagnostic value of non-invasive model for NASH was evaluated by receiver operating characteristic (ROC) curve. Results:The median age in patients were (32±10) years old with a body mass index of (39.16±6.58) kg/m2, including 236 females (63.6%). Serum SHBG level [ M ( Q1, Q3)] in NAFLD group was significantly lower than that in control group [16.90 (11.43, 23.00) vs. (23.45 (15.40, 31.22) mmol/L, P<0.05], and progressively diminished in NAFL, uncertain-NASH and NASH subgroups [(22.24±10.47), (20.57±19.58), (15.80±8.74) mmol; P for trend<0.05]. Compared with the high-leveled SHBG subgroup, the steatosis score (2.09±0.80 vs. 1.51±0.72, P<0.01) and lobular inflammation score (1.10±0.68 vs. 0.85±0.68, P<0.05) were significantly higher in the low-leveled SHBG group. Multivariate logistic regression analysis indicated that lower serum SHBG level was an independent risk factor for NASH ( OR=2.527, 95% CI: 1.296 to 4.928, P<0.05). The area under ROC curve of SHBG combined with aspartate aminotransferase in predicting NASH in NAFLD patients was 0.752 (95% CI: 0.696 to 0.809). Conclusion:Low serum SHBG level is associated with NASH.

Objective:To investigate the prevalences of overweight and obesity and its relationship with the risk factors of cardiovascular diseases among physical examination population in Nanjing Drum Tower Hospital.Methods:A retrospective analysis was performed on 384 061 adults aged 20 years and older. Age- and sex-standardized prevalences of overweight and obesity were calculated using Chinese census data in 2010. Multivariate logistic regression analysis was used to assess the associations of overweight and obesity with the risk factors of cardiovascular diseases.Results:The age-standardized prevalences of overweight and obesity were 42.8% and 13.2% in men and 23.9% and 6.6% in women. A gradually increasing trend was observed in the prevalences of overweight and obesity from 2008 to 2016, especially in subjects aged 20-39 years. Overweight and obesity were significantly associated with increased risks of dyslipidemia, diabetes mellitus, hypertension, and hyperuricemia. These associations were found to be the strongest among subjects aged 20-39 years, which became weaker along with the increasing of age. The OR(95% CI) of dyslipidemia, diabetes mellitus, hypertension, and hyperuricemia were 4.23(4.01-4.47), 3.70(2.97-4.60), 6.19(5.76-6.64), and 3.66(3.45-3.88) in obese men aged 20-39 years, while 5.29(4.63-6.04), 6.38(3.86-10.55), 9.36(7.86-11.13), and 6.65(5.70-7.74) in obese women aged 20-39 years, respectively. Conclusion:The increasing trend in the prevalence of overweight and obesity is a risk factor for cardiovascular diseases in Nanjing adults, especially in individuals aged 20-39 years.

Objective:To investigate the factors influencing the prognosis of talar neck fractures of Hawkins type Ⅲ.Methods:A retrospective analysis was performed of the 18 patients with complete follow-up data who had been treated from October 2013 to December 2016 at Department of Orthopedic Trauma, Beijing Jishuitan Hospital for talar neck fractures of Hawkins type Ⅲ. They were 14 males and 4 females with an average age of 35.1 years (range, from 17 to 50 years). There were 3 open fractures. All the patients were treated with one-stage open reduction and internal fixation. The American Orthopaedic Foot and Ankle Society (AOFAS) scoring system was used to evaluate the functions of ankle and hindfoot during follow-up. The correlations were analyzed between the ankle-hindfoot AOFAS scores and the factors which might have influenced the prognosis, like gender, age, smoking, alcoholic drinking, body mass index, injury cause, medial malleolus osteotomy/fracture, combined fracture, open injury, interval from injury to surgery, operation time, implants (cannulated/plate or Kirschner wire) and infection.Results:The 18 patients were followed up for 21 to 41 months (average, 29.3 months). Of them, 3 had superficial infection which responded to anti-inflammatory therapy and 3 had deep infection which was improved after debridement in 2 cases and led to eventual removal of the talus in one. Tibiotalar arthritis occurred in 6 patients and subtalar arthritis in 10. The AOFAS ankle-hindfoot scores at the last follow-up for this cohort ranged from 12 to 96 points (average, 63.4 points). Ischemic necrosis of the talus was observed in 6 cases and talar collapse in one. The ankle-hindfoot AOFAS scores were strongly correlated with open fracture ( P=0.015,) operation time ( P=0.001), Kirschner wire fixation ( P=0.006), and postoperative infection ( P=0.008). Conclusions:Since open fracture, operation time ≥3 h, Kirschner wire fixation and postoperative infection may be the significant factors that may influence the prognosis of talar neck fractures of Hawkins type Ⅲ, reasonable treatment protocols can help improve the outcomes.