Objective To construct a Pnpla3 148M/M Tm6sf2 167K/K double mutant mouse model by crossbreeding Pnpla3 148M/M homozygous mice and Tm6sf2 167K/K homozygous mice. Methods Pnpla3 148I/M Tm6sf2 167E/K heterozygous mice were bred by hybridization of Pnpla3 148M/M Tm6sf2 167E/E and Pnpla3 148I/I Tm6sf2 167K/K homozygous mice, and the Pnpla3 148M/M Tm6sf2 167K/K mice were obtained by the self-crossbreeding of Pnpla3 148I/M Tm6sf2 167E/K mice. Male mice of Pnpla3 148M/M Tm6sf2 167K/K ( n =6), Pnpla3 148M/M Tm6sf2 167E/E ( n =6), and Pnpla3 148I/I Tm6sf2 167K/K ( n =6) genotypes and Wt mice ( n =6) were fed with normal diet for 8 weeks, and then the glucose and lipid metabolism indices were measured. A one-way analysis of variance was used for comparison between multiple groups, and the least significant difference t -test was used for further comparison bewteen two groups. Results Agarose gel electrophoresis and nucleic acid sequencing results showed that the Pnpla3 148M/M Tm6sf2 167K/K double mutant mouse model was successfully constructed. There were no significant difference in body weight between the Pnpla3 148M/M Tm6sf2 167K/K mice and the Pnpla3 148M/M Tm6sf2 167E/E , Pnpla3 148I/I Tm6sf2 167K/K , and Wt mice (all P > 0.05). The Pnpla3 148M/M Tm6sf2 167K/K mice had a significantly higher liver wet weight than the Wt mice ( P < 0.05). The fasting blood glucose of Pnpla3 148M/M Tm6sf2 167K/K mice was significantly lower than that of Pnpla3 148I/I Tm6sf2 167K/K mice and Wt mice (both P < 0.05). The glucose tolerance of Pnpla3 148M/M Tm6sf2 167K/K mice was significantly reduced compared with the Pnpla3 148I/I Tm6sf2 167K/K mice ( P < 0.05). There were no significant differences in insulin level between the four groups of mice (all P > 0.05). Also, there were no significant differences in the serum levels of biochemical indices between the Pnpla3 148M/M Tm6sf2 167K/K mice and the Pnpla3 148M/M Tm6sf2 167E/E , Pnpla3 148I/I Tm6sf2 167K/K , and Wt mice (all P > 0.05). Oil red O staining of the liver showed that more lipid accumulation was observed in the Pnpla3 148M/M Tm6sf2 167K/K mice than in the Pnpla3 148M/M Tm6sf2 167E/E and Wt mice. Conclusion The Pnpla3 148M/M Tm6sf2 167K/K double mutant mouse model was successfully constructed. Pnpla3 Ⅰ 148M and Tm6sf2 E 167K double mutations can cause abnormal glucose metabolism in mice.

Background: Currently available guidelines contain conflicting recommendations on the management of blood pressure (BP) in patients with diabetes mellitus (DM). Therefore, it is necessary to appraise the guidelines and summarize the agreements and differences among recommendations. Methods: Four databases and the websites of guideline organizations were searched for guidelines regarding BP targets and thresholds for pharmacologic therapy in DM patients, and the included guidelines were appraised with the Appraisal of Guidelines for Research and Evaluation (AGREE) II instrument. Results: In 6,498 records identified, 20 guidelines met our inclusion criteria with 64.0% AGREE II scores (interquartile range, 48.5% to 72.0%). The scores of the European and American guidelines were superior to those of the Asian guidelines (both adjusted P<0.001). Most of the guidelines advocated systolic BP targets <130 mm Hg (12 guidelines, 60%) and diastolic BP targets <80 mm Hg (14 guidelines, 70%) in DM patients. Approximately half of the guidelines supported systolic BP thresholds >140 mm Hg (10 guidelines, 50%) and diastolic BP thresholds >90 mm Hg (nine guidelines, 45%). The tiny minority of the guidelines provided the relevant recommendations regarding the lower limit of official BP targets and the ambulatory BP monitoring (ABPM)/home BP monitoring (HBPM) targets and thresholds in DM patients. Conclusion The lower official BP targets (<130/80 mm Hg) in patients with DM are advocated by most of the guidelines, but they contain conflicting recommendations on the official BP thresholds. Moreover, the gaps regarding the lower limit of official BP targets and the ABPM/HBPM targets and thresholds need to be considered by future study.

A total of three early gastric cancers (EGC)patients with submucosal severe fibrosis were successfully treated by endoscopic precise muscular dissection (EPMD) from May 2018 to August 2020.All the three lesions were en bloc resection and specimens were not damaged. The maximum diameter of specimens and fibrosis areas were 2.0-5.7 cm and 0.5-0.8 cm, respectively. The operation time was from 30 to 70 mins. Intraoperative perforation occurred in one case, post-operation fever cccured in one case, and no delayed perforation or bleeding occured. Two cases were cured resection and one case was non-cured resection without additional treatment. No recurrence or metastasis was observed during follow-up period.According to primary results, EPMD can be used for EGC with submucosal severe fibrosis, but the safety and effectiveness need clinical validation with more samples.

Objective To establish a mouse model of hepatocyte-specific TM6SF2 knockout, and to investigate the role of TM6SF2 in the development of nonalcoholic fatty liver disease (NAFLD). Methods The CRISPR/Cas9 technique and the Cre/LoxP strategy were used to establish a stable mouse model of hepatocyte-specific TM6SF2 knockout. The mice with hepatocyte-specific TM6SF2 knockout and the control mice were given a normal diet or a high-fat diet (HFD) for 16 weeks, and related indices were measured, including general status (body weight and liver weight), glucose metabolic indices (fasting blood glucose and insulin), and lipid metabolism (plasma triglyceride, cholesterol, and liver triglyceride). The t -test was used for comparison of normally distributed continuous data between two groups. Results Under the condition of HFD, compared with the control mice, the mice with hepatocyte-specific TM6SF2 knockout had significantly higher liver weight (2.235±0.175 g vs 1.258±0.106 g, t =4.789, P < 0.01) and liver index (4.970%±0.298% vs 3.210%±0.094%, t =5.630, P < 0.01), and the loss of the TM6SF2 gene in hepatocytes aggravated the abnormal level of alanine aminotransferase induced by HFD (62.517±1.526 U/L vs 25.991±5.947 U/L, t =5.949, P < 0.01). Compared with the control mice under the condition of normal diet or HFD, the mice with TM6SF2 knockout had a significant increase in plasma insulin level (normal diet: 37.203±0.836 mIU/L vs 34.835±0.426 mIU/L, t =2.520, P =0.025; HFD: 41.093±1.226 mIU/L vs 35.817±0.500 mIU/L, t =3.985, P =0.007), while there were no significant differences in the other indices associated with glucose metabolism (all P > 0.05). Under the condition of HFD, there were no significant differences in the levels of plasma triglyceride and cholesterol between the mice with hepatocyte-specific TM6SF2 knockout and the control group ( P > 0.05), while the mice with hepatocyte-specific TM6SF2 knockout had a significant increase in the level of liver triglyceride compared with the control mice (23.969±0.978 mg/g vs 18.229±1.633 mg/g, t =3.015, P =0.024). Conclusion Hepatocyte-specific knockout of TM6SF2 can aggravate liver lipid accumulation and liver injury in mice with NAFLD.

Background: Currently available guidelines contain conflicting recommendations on the management of blood pressure (BP) in patients with diabetes mellitus (DM). Therefore, it is necessary to appraise the guidelines and summarize the agreements and differences among recommendations. Methods: Four databases and the websites of guideline organizations were searched for guidelines regarding BP targets and thresholds for pharmacologic therapy in DM patients, and the included guidelines were appraised with the Appraisal of Guidelines for Research and Evaluation (AGREE) II instrument. Results: In 6,498 records identified, 20 guidelines met our inclusion criteria with 64.0% AGREE II scores (interquartile range, 48.5% to 72.0%). The scores of the European and American guidelines were superior to those of the Asian guidelines (both adjusted P<0.001). Most of the guidelines advocated systolic BP targets <130 mm Hg (12 guidelines, 60%) and diastolic BP targets <80 mm Hg (14 guidelines, 70%) in DM patients. Approximately half of the guidelines supported systolic BP thresholds >140 mm Hg (10 guidelines, 50%) and diastolic BP thresholds >90 mm Hg (nine guidelines, 45%). The tiny minority of the guidelines provided the relevant recommendations regarding the lower limit of official BP targets and the ambulatory BP monitoring (ABPM)/home BP monitoring (HBPM) targets and thresholds in DM patients. Conclusion The lower official BP targets (<130/80 mm Hg) in patients with DM are advocated by most of the guidelines, but they contain conflicting recommendations on the official BP thresholds. Moreover, the gaps regarding the lower limit of official BP targets and the ABPM/HBPM targets and thresholds need to be considered by future study.

Objective:To propose a method of cervical cytology screening based on deep convolutional neural network and compare it with the diagnosis of cytologists.Method:The deep segmentation network was used to extract 618 333 regions of interest (ROI) from 5, 516 cytological pathological images. Combined with the experience of physicians, the deep classification network with the ability to analyze ROI was trained. The classification results were used to construct features, and the decision model was used to complete the classification of cytopathological images.Results:The sensitivity and specificity were 89.72%, 58.48%, 33.95% and 95.94% respectively. Among the smears derived from four different preparation methods, this algorithm had the best effect on natural fallout with a sensitivity of 91.10%, specificity of 69.32%, positive predictive rate of 41.41%, and negative predictive rate of 97.03%.Conclusion:Deep convolutional neural network image recognition technology can be applied to cervical cytology screening.

Three hundred and nine patients with thyroid nodules detected by physical examination in Harrison International Peace Hospital from October 2013 to October 2017 were divided into intervention group (155 cases) and control group (154 cases). Patients in intervention group received oral levothyroxine sodium 25 g / d for 12 months and those in control group had no treatment, patients were followed up every 3 months to 12 months. After treatment, the maximum diameter and thyroid nodule volume of the intervention group were (31.87±3.84) mm and (17.32±0.94) cm3, which were significantly smaller than those of the control group [(34.01±3.72) mm and (24.25±1.21)cm3, P<0.05]. TSH in intervention group was lower than that in control group [(2.24±0.41) vs. (2.52±0.58) mIU/L, P<0.05] and free T4 (FT4) was higher than that in control group [(25.64 ± 3.85) vs. (16.39 ± 3.28) pmol/ L, P<0.05]. TC, TG and LDL?C in intervention group, were lower than those in the control group .The HDL?C level in intervention group was higher than that in control group (all P<0.05). After treatment, there were no malignant changes in the intervention group, while the malignant change rate in control group was 2.6％ (4/154). It is suggested that levothyroxine treatment can reduce TSH level in patients with benign thyroid nodules, inhibit the growth of thyroid nodules.

Objective: To investigate the clinicopathological features, diagnosis and differential diagnosis of dedifferentiated liposarcoma (DDLPS) with inflammatory myofibroblastic tumor (IMT)-like features. Methods: Five cases of DDLPS with IMT-like features were collected from the First Affiliated Hospital of Nanjing Medical University, the Affiliated Hospital of Nanjing University of Traditional Chinese Medicine and the First People′s Hospital of Qinzhou between 2013 and 2018. EnVision method and fluorescence in situ hybridization (FISH) were used to detect the immunophenotype of the tumor cells and the profile of MDM2 gene amplification respectively. Results: All five cases were male and the median age was 61 (range 53 to 65) years. The clinical symptoms were mainly related to the space-occupying lesions. The tumors were located in duodenal mesentery (two cases), intestinal wall (one case), retroperitoneum (one case), and spermatic cord (one case). Grossly, the tumors were not well encapsulated, ranging from 3 to 13 cm (median 6.7 cm) in diameter, with tan to gray and firm cut surface. Histologically, the dedifferentiated component closely resembled inflammatory myofibroblastic tumor (IMT), with spindle/polygonal/stellate-shaped cells arranged in storiform, sheet-like, or random pattern, with varying degrees of chronic inflammation and fibrosis. All three major patterns seen in IMT (myxoid, cellular and hypocellular fibrous) were observed, the hypocellular fibrous pattern was the most common. Well-differentiated liposarcomatous component was found in the peripheral areas of all the tumors. One case had high grade dedifferentiated component. Four cases were strongly positive for MDM2 and p16. Two cases were positive for SMA, and one case was focally positive for desmin and one for CD34. None of the cases stained for ALK-1. FISH demonstrated MDM2 gene amplification in all five cases. Clinical follow-ups were available in all five cases and the interval ranged from 3 to 66 months (median 23 months). Two patients developed recurrences and one patient had metastasis. The remaining two patients were alive with no evidence of tumor recurrence at 3 and 14 months after surgery respectively. Conclusions DDLPS with IMT-like features is a more aggressive neoplasm than its histological mimic (IMT), and should not be misdiagnosed as other intermediate or low-grade malignant tumors, such as IMT, sclerosing liposarcoma, inflammatory liposarcoma, aggressive fibromatosis, solitary fibrous tumors, low-grade myofibroblastic sarcoma, and low-grade fibrosarcoma.

Objective To investigate the clinicopathological features, diagnosis and differential diagnosis of dedifferentiated liposarcoma (DDLPS) with inflammatory myofibroblastic tumor (IMT)?like features. Methods Five cases of DDLPS with IMT?like features were collected from the First Affiliated Hospital of Nanjing Medical University, the Affiliated Hospital of Nanjing University of Traditional Chinese Medicine and the First People′s Hospital of Qinzhou between 2013 and 2018. EnVision method and fluorescence in situ hybridization (FISH) were used to detect the immunophenotype of the tumor cells and the profile of MDM2 gene amplification respectively. Results All five cases were male and the median age was 61 (range 53 to 65) years. The clinical symptoms were mainly related to the space?occupying lesions. The tumors were located in duodenal mesentery (two cases), intestinal wall (one case), retroperitoneum (one case), and spermatic cord (one case). Grossly, the tumors were not well encapsulated, ranging from 3 to 13 cm (median 6.7 cm) in diameter, with tan to gray and firm cut surface. Histologically, the dedifferentiated component closely resembled inflammatory myofibroblastic tumor (IMT), with spindle / polygonal /stellate?shaped cells arranged in storiform, sheet?like, or random pattern, with varying degrees of chronic inflammation and fibrosis. All three major patterns seen in IMT (myxoid, cellular and hypocellular fibrous) were observed, the hypocellular fibrous pattern was the most common. Well?differentiated liposarcomatous component was found in the peripheral areas of all the tumors. One case had high grade dedifferentiated component. Four cases were strongly positive for MDM2 and p16. Two cases were positive for SMA, and one case was focally positive for desmin and one for CD34. None of the cases stained for ALK?1. FISH demonstrated MDM2 gene amplification in all five cases. Clinical follow?ups were available in all five cases and the interval ranged from 3 to 66 months (median 23 months). Two patients developed recurrences and one patient had metastasis. The remaining two patients were alive with no evidence of tumor recurrence at 3 and 14 months after surgery respectively. Conclusions DDLPS with IMT?like features is a more aggressive neoplasm than its histological mimic (IMT), and should not be misdiagnosed as other intermediate or low?grade malignant tumors, such as IMT, sclerosing liposarcoma, inflammatory liposarcoma, aggressive fibromatosis, solitary fibrous tumors, low?grade myofibroblastic sarcoma, and low?grade fibrosarcoma.

Objective To investigate the mutations of IDH1,IDH2,p53 gene,and Ki-67 protein expression in different grade of gliomas and identify the association with its clinical relevance. Methods The mutations of IDH1,IDH2 and p53 gene were detected by direct DNA sequencing,and protein expression of Ki-67 was analyzed by immunohistochemistry. The correlations between gender,age,tumor site,differentiation degree and pathological type of patients were analyzed. Results R132H mutation of IDH1 gene was detected in 32.6% samples (14/46 cases),of which the proportion of WHO classification grade Ⅱ was 40.0%,and grade Ⅲ was 58.3%. IDH1 mutations were shown correlated with age,pathology level Ⅱ-Ⅲ,and Ki-67 low expression. p53 mutations were detected in 4 glioblastomas,with mutations located at exon 7,8. IDH1 gene mutation was negatively correlated with Ki-67 expression. Conclusions The proportion of IDH1 gene mutation in different pathological types of gliomas is different,which is the highest in gradeⅡ~Ⅲ. It is suggested that the subtypes should be listed independently by routine tests. Mutations in p53 gene are more common in primary glioblastomas and may be associated with adverse outcomes. The combined detection of DH1,p53 and Ki-67 is conducive to the diagnosis and prognosis of glioma.