Objective:To investigate the clinicopathological features including immunophenotype, molecular characteristics, differential diagnosis and prognosis of SMARCB1-deficient renal medullary carcinoma (RMC) without sickle cell trait.Methods:The clinicopathological data of 12 cases of SMARCB1-deficient RMC without sickle cell trait were collected from 7 domestic institutions during the period of 2015 to 2024. Their clinical characteristics, morphological features and immunohistochemical properties were observed and analyzed. High-throughput DNA-targeted next-generation sequencing was performed, and follow-up data were gathered along with relevant literature review.Results:Among the 12 patients, 5 were female and 7 were male. The patients age ranged from 27 to 84 years with a median age of 58.5 (46.0, 71.0) years. None of them had sickle cell disease or other hemoglobinopathies. Eight cases occurred in the left kidney and 4 cases were located in the right kidney. The average maximum diameter of the tumor was 6.1 (4.0,7.5) cm, with a range of 2.0 to 14.9 cm (the median maximum diameter 5.5 cm). Histologically, the tumors showed poorly differentiated adenocarcinoma, arranged in solid and tubular patterns. Papillary structure was noted in 5 cases, cribriform structure in 3 cases, rhabdoid differentiation in 3 cases, and sarcomatoid differentiation in 2 cases. Inflammatory desmoplastic stromal reaction was observed in 8 cases, among which stromal myxoid degeneration was seen in 6 cases. Tumor necrosis was apparent in 6 cases. The tumor cells had abundant eosinophilic or clear cytoplasm and prominent nucleoli. The nuclear grading was grade 3 or 4 according to the International Society of Urological Pathology (ISUP). Immunohistochemical staining showed that the tumor cells of all 12 cases expressed PAX8 and loss of SMARCB1/INI1 protein expression, and 5 of 10 cases expressed OCT3/4. Seven samples had valid archived paraffin tissues for high-throughput DNA-targeted next-generation sequencing. The results showed that all 7 cases had pathogenic mutations in the SMARCB1 gene. The mutation sites included exon5 c.595A>T (p.K199*), exon2 c.200_207del (p.S67*), exon2 p.G69VfsTer16, exon7 c.986G>T (p.S329I), exon7 c.886A>T (p.K296*), exon6 c.635T>A (p.L212*), exon5 c.577del (p.M193Wfs16), and exon6 c.784del (p.V262Sfs5). Follow-up data were obtained for 6 of 12 patients. Among them, 1 patient had lung and bone metastases, 1 patient had liver and bone metastases and 1 patient had multiple bone metastases at the time of diagnosis; 1 patient had bone metastases 5 months after surgery. One patient died of postoperative complications 10 days after surgery, 4 patients died of tumors (the survival time ranged from 4 to 8 months), and 1 patient had no recurrence or metastasis during the 8-month follow-up after surgery.Conclusions:SMARCB1-deficient RMC without sickle cell trait is a highly aggressive and poorly differentiated renal cell carcinoma. It has similar histomorphology, immunophenotype, molecular characteristics and prognosis to RMC, which further supports that it is a sporadic subtype of RMC related to sickle cell trait.

Objective:To investigate the characteristics of crystalline lens tilt and decentration distribution in age-related cataract patients and analyze the related factors.Methods:A cross-sectional study was conducted.A total of 585 consecutive patients (1 170 eyes) with bilateral age-related cataracts over the age of 50 were enrolled at Aier Eye Hospital of Wuhan University from January 2022 to March 2022.Bilateral crystalline lens tilt and decentration magnitude and direction were measured by swept-source anterior segment optical coherence tomography.Corneal curvature, white-to-white corneal diameter, anterior chamber depth, lens thickness and axial length were measured using a swept-source optical coherence tomography biometer.The correlation of tilt magnitude and decentration magnitude of crystalline lens was assessed by Pearson linear correlation analysis.The correlation of tilt direction and decentration direction of crystalline lens was assessed by Spearman rank correlation analysis.The influence of age and ocular biometric parameters on crystalline lens tilt and decentration was analyzed by multiple linear regression analysis.This study adhered to the Declaration of Helsinki and was approved by the Ethics Committee of Aier Eye Hospital of Wuhan University (No.2022IRBKY0617).All patients signed the informed consent form before any medical examination.Results:The mean tilt magnitude was (4.69±1.59)°, of which 6.7% had a tillt ≥7°, and the direction was mainly toward the inferotemporal side with a mirror symmetry between both eyes.The mean decentration magnitude was (0.19±0.11)mm, of which 5.3% had a decentration ≥0.4 mm, and the direction was mainly towards the temporal side.The crystalline lens tilt magnitude was positively correlated with decentration magnitude ( r=0.422, P<0.001) and the crystalline lens tilt direction was positively correlated with decentration direction ( rs=0.432, P<0.001).There was no significant difference in crystalline lens tilt magnitude between both eyes ( t=0.487, P=0.626), and the crystalline lens decentration was significantly greater in the right eye than in the left eye, with statistically significant difference ( t=2.563, P=0.011).There was no significant difference in crystalline lens tilt and decentration between male and female ( t=1.500, 1.566; both P>0.05).The crystalline lens tilt magnitude was significantly greater in 60-69 years, 70-79 years and 80-89 years groups than in 50-59 years group, and the crystalline lens decentration magnitude was significantly greater in the 80-89 years group than in the 60-69 years group (all P<0.05).The crystalline lens tilt magnitude was smaller in the axial length ≥25 mm group than in the <22 mm group and the 22-<25 mm group, and the differences were statistically significant (both P<0.001).There was no significant difference in crystalline lens decentration magnitude among different axial length groups ( F=1.433, P=0.239).The multiple linear regression analysis revealed that the effects of anterior chamber depth ( B=-0.474, t=-2.115, P=0.035), axial length ( B=-0.258, t=-9.324, P<0.001) and crystalline lens decentration ( B=5.646, t=11.234, P<0.001) on crystalline lens tilt were statistically significant.The influence of lens thickness ( B=-0.032, t=-2.402, P=0.017), axial length ( B=0.010, t=4.406, P<0.001) and crystalline lens tilt ( B=0.032, t=11.234, P<0.001) on crystalline lens decentration were statistically significant. Conclusions:Among age-related cataract patients aged 50 and above, 6.7% have crystalline lens tilt ≥7°, and 5.3% have crystalline lens decentration ≥0.4 mm.The older the age, the greater the crystalline lens tilt and decentration.The shorter the axial length and the shallower the anterior chamber, the greater the tilt of the lens.The greater the tilt magnitude and decentration, the greater the decentration magnitude and decentration.

Purpose To investigate the clinicopathological features and possible tumor-associated immune micro-environment in CD23-positive diffuse large B-cell lymphoma(DLBCL).Methods The clinicopathological data of 12 cases of CD23-positive DLBCL patients were analyzed retrospectively.The clinical and pathological features were ana-lyzed,and the clinical correlation and tumor-associated immune invasion were studied.Results CD23-positive DL-BCL accounted for 9.45％of all DLBCL.There were 6 males and 6 females.The mean age of onset was 64.83 years old.Four DLBCL cases occurred in lymph nodes and 8 cases occurred outside lymph nodes.Nine DLBCL cases were in advanced stage(Ⅲ-Ⅳ)and 3 cases DLBCL were in early stage(Ⅰ-Ⅱ).Among the patients,3 cases were untreated and lost to follow-up.One case deteriorated and died after operation.Two cases died,1 case progressed and 5 cases partially recovered after chemotherapy.Microscopically,the tumor cells were diffusely infiltrated and destroyed the nor-mal tissue structure.The tumor cells were observed to be centroblastic,immunoblastic and anaplastic large cells.No blastoid transformation and plasmacytoid differentiation were observed in morphology.According to Hans algorithm,11 cases were non-GCB phenotype except 1 case was GCB phenotype.Bioinformatics studies revealed that CD23 expres-sion was correlated with regulatory T cells,NK cells,plasma-like dendritic cells and neutrophils.Conclusion CD23-positive DLBCL patients are mainly middle-aged and elderly,and most of them occur outside lymph nodes and in ad-vanced stage(Ⅲ-Ⅳ).Follow-up results show that their prognosis is poor.Morphologically,there is no significant difference between DLBCL and conventional DLBCL.The Hans classification suggests that most cases originated from activated B cells.CD23 expression may play a role in the immune microenvironment of DLBCL.

Objective:To investigate the characteristics of crystalline lens tilt and decentration distribution in age-related cataract patients and analyze the related factors.Methods:A cross-sectional study was conducted.A total of 585 consecutive patients (1 170 eyes) with bilateral age-related cataracts over the age of 50 were enrolled at Aier Eye Hospital of Wuhan University from January 2022 to March 2022.Bilateral crystalline lens tilt and decentration magnitude and direction were measured by swept-source anterior segment optical coherence tomography.Corneal curvature, white-to-white corneal diameter, anterior chamber depth, lens thickness and axial length were measured using a swept-source optical coherence tomography biometer.The correlation of tilt magnitude and decentration magnitude of crystalline lens was assessed by Pearson linear correlation analysis.The correlation of tilt direction and decentration direction of crystalline lens was assessed by Spearman rank correlation analysis.The influence of age and ocular biometric parameters on crystalline lens tilt and decentration was analyzed by multiple linear regression analysis.This study adhered to the Declaration of Helsinki and was approved by the Ethics Committee of Aier Eye Hospital of Wuhan University (No.2022IRBKY0617).All patients signed the informed consent form before any medical examination.Results:The mean tilt magnitude was (4.69±1.59)°, of which 6.7% had a tillt ≥7°, and the direction was mainly toward the inferotemporal side with a mirror symmetry between both eyes.The mean decentration magnitude was (0.19±0.11)mm, of which 5.3% had a decentration ≥0.4 mm, and the direction was mainly towards the temporal side.The crystalline lens tilt magnitude was positively correlated with decentration magnitude ( r=0.422, P<0.001) and the crystalline lens tilt direction was positively correlated with decentration direction ( rs=0.432, P<0.001).There was no significant difference in crystalline lens tilt magnitude between both eyes ( t=0.487, P=0.626), and the crystalline lens decentration was significantly greater in the right eye than in the left eye, with statistically significant difference ( t=2.563, P=0.011).There was no significant difference in crystalline lens tilt and decentration between male and female ( t=1.500, 1.566; both P>0.05).The crystalline lens tilt magnitude was significantly greater in 60-69 years, 70-79 years and 80-89 years groups than in 50-59 years group, and the crystalline lens decentration magnitude was significantly greater in the 80-89 years group than in the 60-69 years group (all P<0.05).The crystalline lens tilt magnitude was smaller in the axial length ≥25 mm group than in the <22 mm group and the 22-<25 mm group, and the differences were statistically significant (both P<0.001).There was no significant difference in crystalline lens decentration magnitude among different axial length groups ( F=1.433, P=0.239).The multiple linear regression analysis revealed that the effects of anterior chamber depth ( B=-0.474, t=-2.115, P=0.035), axial length ( B=-0.258, t=-9.324, P<0.001) and crystalline lens decentration ( B=5.646, t=11.234, P<0.001) on crystalline lens tilt were statistically significant.The influence of lens thickness ( B=-0.032, t=-2.402, P=0.017), axial length ( B=0.010, t=4.406, P<0.001) and crystalline lens tilt ( B=0.032, t=11.234, P<0.001) on crystalline lens decentration were statistically significant. Conclusions:Among age-related cataract patients aged 50 and above, 6.7% have crystalline lens tilt ≥7°, and 5.3% have crystalline lens decentration ≥0.4 mm.The older the age, the greater the crystalline lens tilt and decentration.The shorter the axial length and the shallower the anterior chamber, the greater the tilt of the lens.The greater the tilt magnitude and decentration, the greater the decentration magnitude and decentration.

Purpose To investigate the clinicopathological features and possible tumor-associated immune micro-environment in CD23-positive diffuse large B-cell lymphoma(DLBCL).Methods The clinicopathological data of 12 cases of CD23-positive DLBCL patients were analyzed retrospectively.The clinical and pathological features were ana-lyzed,and the clinical correlation and tumor-associated immune invasion were studied.Results CD23-positive DL-BCL accounted for 9.45％of all DLBCL.There were 6 males and 6 females.The mean age of onset was 64.83 years old.Four DLBCL cases occurred in lymph nodes and 8 cases occurred outside lymph nodes.Nine DLBCL cases were in advanced stage(Ⅲ-Ⅳ)and 3 cases DLBCL were in early stage(Ⅰ-Ⅱ).Among the patients,3 cases were untreated and lost to follow-up.One case deteriorated and died after operation.Two cases died,1 case progressed and 5 cases partially recovered after chemotherapy.Microscopically,the tumor cells were diffusely infiltrated and destroyed the nor-mal tissue structure.The tumor cells were observed to be centroblastic,immunoblastic and anaplastic large cells.No blastoid transformation and plasmacytoid differentiation were observed in morphology.According to Hans algorithm,11 cases were non-GCB phenotype except 1 case was GCB phenotype.Bioinformatics studies revealed that CD23 expres-sion was correlated with regulatory T cells,NK cells,plasma-like dendritic cells and neutrophils.Conclusion CD23-positive DLBCL patients are mainly middle-aged and elderly,and most of them occur outside lymph nodes and in ad-vanced stage(Ⅲ-Ⅳ).Follow-up results show that their prognosis is poor.Morphologically,there is no significant difference between DLBCL and conventional DLBCL.The Hans classification suggests that most cases originated from activated B cells.CD23 expression may play a role in the immune microenvironment of DLBCL.

Objective:To investigate the clinicopathological features including immunophenotype, molecular characteristics, differential diagnosis and prognosis of SMARCB1-deficient renal medullary carcinoma (RMC) without sickle cell trait.Methods:The clinicopathological data of 12 cases of SMARCB1-deficient RMC without sickle cell trait were collected from 7 domestic institutions during the period of 2015 to 2024. Their clinical characteristics, morphological features and immunohistochemical properties were observed and analyzed. High-throughput DNA-targeted next-generation sequencing was performed, and follow-up data were gathered along with relevant literature review.Results:Among the 12 patients, 5 were female and 7 were male. The patients age ranged from 27 to 84 years with a median age of 58.5 (46.0, 71.0) years. None of them had sickle cell disease or other hemoglobinopathies. Eight cases occurred in the left kidney and 4 cases were located in the right kidney. The average maximum diameter of the tumor was 6.1 (4.0,7.5) cm, with a range of 2.0 to 14.9 cm (the median maximum diameter 5.5 cm). Histologically, the tumors showed poorly differentiated adenocarcinoma, arranged in solid and tubular patterns. Papillary structure was noted in 5 cases, cribriform structure in 3 cases, rhabdoid differentiation in 3 cases, and sarcomatoid differentiation in 2 cases. Inflammatory desmoplastic stromal reaction was observed in 8 cases, among which stromal myxoid degeneration was seen in 6 cases. Tumor necrosis was apparent in 6 cases. The tumor cells had abundant eosinophilic or clear cytoplasm and prominent nucleoli. The nuclear grading was grade 3 or 4 according to the International Society of Urological Pathology (ISUP). Immunohistochemical staining showed that the tumor cells of all 12 cases expressed PAX8 and loss of SMARCB1/INI1 protein expression, and 5 of 10 cases expressed OCT3/4. Seven samples had valid archived paraffin tissues for high-throughput DNA-targeted next-generation sequencing. The results showed that all 7 cases had pathogenic mutations in the SMARCB1 gene. The mutation sites included exon5 c.595A>T (p.K199*), exon2 c.200_207del (p.S67*), exon2 p.G69VfsTer16, exon7 c.986G>T (p.S329I), exon7 c.886A>T (p.K296*), exon6 c.635T>A (p.L212*), exon5 c.577del (p.M193Wfs16), and exon6 c.784del (p.V262Sfs5). Follow-up data were obtained for 6 of 12 patients. Among them, 1 patient had lung and bone metastases, 1 patient had liver and bone metastases and 1 patient had multiple bone metastases at the time of diagnosis; 1 patient had bone metastases 5 months after surgery. One patient died of postoperative complications 10 days after surgery, 4 patients died of tumors (the survival time ranged from 4 to 8 months), and 1 patient had no recurrence or metastasis during the 8-month follow-up after surgery.Conclusions:SMARCB1-deficient RMC without sickle cell trait is a highly aggressive and poorly differentiated renal cell carcinoma. It has similar histomorphology, immunophenotype, molecular characteristics and prognosis to RMC, which further supports that it is a sporadic subtype of RMC related to sickle cell trait.

Objective:To observe the ameliorative effects of exogenous miR-146a-5p on lipopolysaccharide (LPS)-induced embryonic resorption and fetal mouse dysplasiamice, and to preliminarily investigate its mechanism of action.Methods:1) After 36 healthy adult female mice were mated with male mice, uterine tissues were collected from females on day (D) 0 (D0/not pregnant), D0.5 (the day of embryo observed), D4.5, D7.5, D9.5 and D13.5 of gestation, and the expression levels of miR-146a-5p and its target gene TRAF6 protein in uterine tissues of mice at different gestation periods were detected by real-time fluorescent quantitative PCR (qPCR) and Western blotting. 2) The mice on D7.5 of pregnancy were treated with intraperitoneal injection of saline (control, COL group), intraperitoneal injection of 250 μg/kg LPS (named LPS250 group), LPS combined with tail vein injection of 10 nmol miR-146a-5p unrelated sequence (negative control, NC, named LPS250+NC group), or LPS combined with tail vein injection of 10 nmol miR-146a-5p agonist (miR-146a-5p agomir, named LPS250+miR-146a-5p agomir group). The total number of embryos and the number of absorbed embryos in the uterus of pregnant mice were measured and statistically analyzed on D8.5, and the expression levels of TNFα mRNA and TRAF6 protein in uterine tissues were detected by qPCR and Western blotting. 3) Then we reduced the dosage of LPS to 50 μg/kg and treated the same groups, named LPS50+NC group, LPS50+miR-146a-5p agomir group, respectively. The total number of fetal mice/embryos, the number of absorbed embryos, the number of surviving fetal mice, the weight of surviving fetal mice and the weight of the placenta were measured and statistically analyzed on D16.5. 4) Primary mouse bone marrow-derived macrophages (BMDM) were isolated and cultured. Mouse BMDM was inducted to M1 polarization by LPS stimulation, and then was transient transfected miR-146a-5p mimics or their NC fragments. The expression levels of TNFα mRNA and pSTAT1 protein were detected by qPCR and Western blotting. Results:The expression level of miR-146a-5p was significantly higher in the implantation sites of D7.5, D9.5 and D13.5 pregnant mice than in the non-implantation sites ( P=0.013, P=0.012, P=0.003), and the protein expression level of TRAF6 was significantly lower in the implantation site of D13.5 pregnant mice than in the non-implantation site ( P=0.012). After intraperitoneal injection of 250 μg/kg of LPS into D7.5 pregnant mice, the embryo absorption rate of the LPS group on D8.5 was 43.13%±3.31%, which was significantly higher than that of COL group (0%, P=0.002), while the embryo absorption rate of the LPS250+miR-146a-5p agomir group (13.50%±0.87%) was significantly lower than that of the LPS250+NC group (59.33%±4.04%, P=0.001). After intraperitoneal injection of 50 μg/kg of LPS combined with tail vein injection of NC or miR-146a-5p agomir to D7.5 pregnant mice, the fetal mouse weight [(0.29±0.09) g] and placental weight [(0.06±0.02) g] of surviving fetal mice in the LPS50+NC group on D16.5 and the LPS50+miR-146a-5p agomir group were statistically significant [(0.46±0.06) g, P<0.001; (0.07±0.02) g, P=0.021], and the differences in the number of absorbed embryos and embryo uptake rate between the two groups were not statistically significant (all P>0.05). The expression levels of both pSTAT1 protein and TNFα mRNA were significantly downregulated in BMDM transfected with miR-146a-5p mimics compared with those transfected with NC ( P=0.012, P=0.039). Conclusion:miR-146a-5p expression levels were significantly increased at the maternal-fetal interface during the late stage of mouse embryo implantation and placental development. Exogenous miR-146a-5p could effectively improve LPS-induced mouse embryo resorption and fetal mouse dysplasia. miR-146a-5p could inhibit the M1 polarization activity of mouse macrophages, suggesting that miR-146a-5p may inhibit the M1 polarization activity of mouse macrophages by suppressing M1 polarization of mouse maternal-fetal interface macrophages to safeguard the normal establishment and maintenance of pregnancy.

Objective:To observe the ameliorative effects of exogenous miR-146a-5p on lipopolysaccharide (LPS)-induced embryonic resorption and fetal mouse dysplasiamice, and to preliminarily investigate its mechanism of action.Methods:1) After 36 healthy adult female mice were mated with male mice, uterine tissues were collected from females on day (D) 0 (D0/not pregnant), D0.5 (the day of embryo observed), D4.5, D7.5, D9.5 and D13.5 of gestation, and the expression levels of miR-146a-5p and its target gene TRAF6 protein in uterine tissues of mice at different gestation periods were detected by real-time fluorescent quantitative PCR (qPCR) and Western blotting. 2) The mice on D7.5 of pregnancy were treated with intraperitoneal injection of saline (control, COL group), intraperitoneal injection of 250 μg/kg LPS (named LPS250 group), LPS combined with tail vein injection of 10 nmol miR-146a-5p unrelated sequence (negative control, NC, named LPS250+NC group), or LPS combined with tail vein injection of 10 nmol miR-146a-5p agonist (miR-146a-5p agomir, named LPS250+miR-146a-5p agomir group). The total number of embryos and the number of absorbed embryos in the uterus of pregnant mice were measured and statistically analyzed on D8.5, and the expression levels of TNFα mRNA and TRAF6 protein in uterine tissues were detected by qPCR and Western blotting. 3) Then we reduced the dosage of LPS to 50 μg/kg and treated the same groups, named LPS50+NC group, LPS50+miR-146a-5p agomir group, respectively. The total number of fetal mice/embryos, the number of absorbed embryos, the number of surviving fetal mice, the weight of surviving fetal mice and the weight of the placenta were measured and statistically analyzed on D16.5. 4) Primary mouse bone marrow-derived macrophages (BMDM) were isolated and cultured. Mouse BMDM was inducted to M1 polarization by LPS stimulation, and then was transient transfected miR-146a-5p mimics or their NC fragments. The expression levels of TNFα mRNA and pSTAT1 protein were detected by qPCR and Western blotting. Results:The expression level of miR-146a-5p was significantly higher in the implantation sites of D7.5, D9.5 and D13.5 pregnant mice than in the non-implantation sites ( P=0.013, P=0.012, P=0.003), and the protein expression level of TRAF6 was significantly lower in the implantation site of D13.5 pregnant mice than in the non-implantation site ( P=0.012). After intraperitoneal injection of 250 μg/kg of LPS into D7.5 pregnant mice, the embryo absorption rate of the LPS group on D8.5 was 43.13%±3.31%, which was significantly higher than that of COL group (0%, P=0.002), while the embryo absorption rate of the LPS250+miR-146a-5p agomir group (13.50%±0.87%) was significantly lower than that of the LPS250+NC group (59.33%±4.04%, P=0.001). After intraperitoneal injection of 50 μg/kg of LPS combined with tail vein injection of NC or miR-146a-5p agomir to D7.5 pregnant mice, the fetal mouse weight [(0.29±0.09) g] and placental weight [(0.06±0.02) g] of surviving fetal mice in the LPS50+NC group on D16.5 and the LPS50+miR-146a-5p agomir group were statistically significant [(0.46±0.06) g, P<0.001; (0.07±0.02) g, P=0.021], and the differences in the number of absorbed embryos and embryo uptake rate between the two groups were not statistically significant (all P>0.05). The expression levels of both pSTAT1 protein and TNFα mRNA were significantly downregulated in BMDM transfected with miR-146a-5p mimics compared with those transfected with NC ( P=0.012, P=0.039). Conclusion:miR-146a-5p expression levels were significantly increased at the maternal-fetal interface during the late stage of mouse embryo implantation and placental development. Exogenous miR-146a-5p could effectively improve LPS-induced mouse embryo resorption and fetal mouse dysplasia. miR-146a-5p could inhibit the M1 polarization activity of mouse macrophages, suggesting that miR-146a-5p may inhibit the M1 polarization activity of mouse macrophages by suppressing M1 polarization of mouse maternal-fetal interface macrophages to safeguard the normal establishment and maintenance of pregnancy.

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.