Objective To investigate the effects of active vitamin D (VD) on the expression of triggering receptor expressed on myeloid cells-1 (TREM-1) in renal tissue of diabetic nephropathies (DN) rats and to explore the impact of TREM-1 on adhesion and migration capacity of macrophage.Methods DN rat models were established by streptozotocin.Rats were randomly distributed into four groups:control (NC) group,VD group,DN group and DN+VD group (DN rats with 0.1 μg · kg-1 · d-1 calcitriol by garages).Rats were sacrificed respectively at 8 weeks and 12 weeks after treatment.Pathological changes in kidney tissue were detected and the expressions of CD68 and TREM-1 were acquired by immunohistochemistry stain and Western blotting.In vitro,RAW264.7 cells were divided into NC group,VD group,high glucose (HG) group and HG+VD group.In HG+VD group rats were treated by high glucose with 10-8 mol/L 1,25(OH)2D3.TREM-1 expression was measured by immunohistochemistry stain and Western blotting,and the ability of macrophage in migration and adhesion was evaluated by Transwell migration assay and adhesion assay.TREM-1 siRNA was transferred to silence TREM-1 expression,while plasmid of TREM-1 was transferred for high expression.Their ability of adhesion and migration in macrophage and the effect of 1,25(OH)2D3 were examined.Results (1) Compared with the NC group,the expressions of CD68 and TREM-1 were increased in DN group (P ＜ 0.05),whereas markedly decreased in DN+VD group (P ＜ 0.05).(2) The number of adhesion and migration cells,and the expression of TREM-1 protein in macrophage were obviously increased in HG group as compared with those in NC group (all P ＜ 0.05);whereas above changes were markedly decreased in HG+VD group than those in HG group (P ＜ 0.05).(3) The number of adhesion and migrated macrophage was reduced after TREM-1 siRNA intervention (all P ＜ 0.05).VD could significantly decrease the effect of high glucose on adhesion and migrated macrophages after TREM-1 siRNA (all P ＜ 0.05).(4) Adhesion and migration of macrophage were increased via TREM-1 overexpression (all P ＜ 0.05),but the effects of VD on high glucose-induced adhesion and migration of macrophage were disappeared.Conclusions VD can suppress the adhesion and migration of macrophage via reducing the expression of TREM-1,and inhibit infiltration of macrophage in renal tissue of DN rats.

Objective To investigate the effect of macrophages on podocytes apoptosis in diabetic nephropathy. Methods Differentiated mouse macrophages (RAW264.7) were exposed to normal glucose, high glucose, then the conditioned media (CM) was collected and considered as NC-CM or HG-CM, respectively. Western blotting and immunofluorescent staining were used to detect the specific markers for M1 macrophages (iNOS) and M2 macrophages (MR). ELISA was used to detect the concentration of TNF-α in the CM. Then normal PRMI 1640 media (control), NC-CM or HG-CM was added to podocytes. In some experiments, ROS inhibitors (Tempo), p38 MAPK inhibitor (SB203580), anti-TNF-α neutralizing antibody, and IgG1 isotype control were respectively added to cells with HG-CM. Besides, recombinant mouse TNF-α alone was applied to incubate podocytes. Podocytes apoptosis was accessed by Annexin V-FITC/PI and Hoechst33342 staining. DCFH-DA staining was used to analyse ROS level. Western blotting was used to detect cleaved casepase-3, p38MAPK, and p-p38MAPK protein. Results Macrophages were activated when exposed to high glucose, displaying pro-inflammatory M1 polarization with higher iNOS and lower MR expression. HG-CM but not NC-CM trigged podocytes apoptosis, up-regulated ROS, cleaved casepase-3 and p-p38MAPK. However, the podocytes apoptosis trigged by HG-CM was abolished by either a ROS inhibitor (Tempo) or a p38 MAPK inhibitor (SB203580). Additionally, TNF-α was increased in the HG-CM. TNF-α protein in macrophage was aslo increased when exposed to high glucose. Anti-TNF-α neutralizing antibody blunted the apoptotic response, excess ROS generation and p-p38 MPAK expression in podocytes induced by HG-CM. Moreover, addition of recombinant TNF-α similarly led to podocytes apoptosis, increased ROS and p38 MPAK expression. Conclusion M1 macrophages activated by high glucose released TNF-α to promote podocytes apoptosis via ROS-p38 MAPK pathway.

Objective:To understand the occupational health examination of graphene workers in a certain enterprise and provide evidence for occupational disease prevention and treatment.Methods:In September 2019, the physical examination data of 54 workers exposed to graphene dust in an occupational health examination of a graphene enterprise from January to August 2019 were collected, and the differences of various physical examination indexes of each group were examined by χ 2 test. Results:The company had a total of 54 graphene workers, and 15 cases of occupational health examination abnormalities, and the abnormal rate was 27.8%. 7 of which had abnormal central functions, and 11 cases of abnormal pulmonary function. 3 of them had abnormal heart function and abnormal lung function. With the increase of age and working age, the abnormal rates of cardiac and pulmonary functions were increased (χ 2=0.042, P<0.05) . Conclusion:With the development and mass production of graphene, the opportunities for workers to expose to graphene are increasing, and the health of workers is also threatened. Therefore, the occupational disease prevention work of staff should be emphasized.

Objective:To understand the occupational health examination of graphene workers in a certain enterprise and provide evidence for occupational disease prevention and treatment.Methods:In September 2019, the physical examination data of 54 workers exposed to graphene dust in an occupational health examination of a graphene enterprise from January to August 2019 were collected, and the differences of various physical examination indexes of each group were examined by χ 2 test. Results:The company had a total of 54 graphene workers, and 15 cases of occupational health examination abnormalities, and the abnormal rate was 27.8%. 7 of which had abnormal central functions, and 11 cases of abnormal pulmonary function. 3 of them had abnormal heart function and abnormal lung function. With the increase of age and working age, the abnormal rates of cardiac and pulmonary functions were increased (χ 2=0.042, P<0.05) . Conclusion:With the development and mass production of graphene, the opportunities for workers to expose to graphene are increasing, and the health of workers is also threatened. Therefore, the occupational disease prevention work of staff should be emphasized.

Objective To train influenza clinical syndrome data using machine learning methods;To obtain an influenza syndrome differentiation model.Methods The medical records of influenza patients who visited the fever clinic of China-Japan Friendship Hospital from December 2019 to March 2022 were collected.The data set system was used for data processing,and the data generated by different data processing processes were stored separately for training.The study selected logistic regression,decision tree,naive Bayes,support vector machine,multi-layer perceptron,lightGBM and random forest as alternative models,and optimized the hyperparameters through Optuna.Models were trained separately in each data set,and the model prediction performance was evaluated,with the macro-F1 score as the core.Results Totally 1 011 training samples were collected,including 453 cases of wind-heat syndrome,152 cases of superficial wind-cold syndrome,and 406 cases of superficial cold and internal heat syndrome;8 data sets were obtained for training,containing 80 copies of data.After training,the macro-F1 scores of logistic regression,decision tree,naive Bayes,support vector machine,multi-layer perceptron lightGBM and random forest model were 0.783 0,0.774 2,0.731 5,0.782 4,0.716 7,0.793 8 and 0.815 3,respectively.Weighted samples could significantly improve the average model performance,while PCA would reduce the average model performance.The prediction performance of the logistic regression model was the best in the single method models,and the random forest model was the best in the integrated method models.Conclusion In the case of a small sample size,it is more appropriate to use logistic regression,decision tree,support vector machine and lightGBM for the TCM influenza syndrome differentiation model.As the sample size increases,logistic regression,support vector machine,lightGBM and random forest may be more suitable.Different data processing methods will affect model performance.Collecting information on the typical degree of syndrome types is beneficial to improving model performance.

Objective To observe the impact factors of the diagnostic accuracy of fine needle aspiration biopsy(FNAB)for papillary thyroid carcinoma(PTC).Methods Totally 468 patients with single PTC confirmed by postoperative pathology who underwent FNAB before surgery were enrolled.The impact of clinica,l ultrasonic and pathological features on the accuracy of FNAB diagnosis were analyzed.Results The accuracy of FNAB for diagnosing PTC was 71.37%(334/468).The maximum diameter and location of PTC were both impact factors of the diagnostic accuracy of FNAB.The maximum diameter of 0.7 cm was the optimal cutoff value of FNAB for diagnosing PTC,and the diagnostic accuracy of FNAB for PTC with the maximum diameter＜0.7 cm and those≥0.7 cm was 62.96%(119/189)and 77.06%(215/279),respectively.The diagnostic accuracy of FNAB for PTC located in the difficult and easy area of puncture was 52.53%(52/99)and 76.42%(282/369),respectively.The diagnostic accuracy of FNAB for PTC with the maximum diameter≥0.7 cm and located in the easy area,≥0.7 cm and located in the difficult area,＜0.7 cm and located in the easy area,＜0.7 cm and located in the difficult area was 80.43%(185/230),61.22%(30/49),69.78%(97/139)and 44.00%(22/50),respectively.Conclusion The maximum diameter and location of PTC were both impact factors of the diagnostic accuracy of FNAB.

Objective: To understand the characteristics and dynamics of individuals with HIV-1 subtype infection among injected drug users (HIV infection IDU) in Guangzhou between 2008 and 2015. Methods: HIV-1 RNAs were extracted from serum samples of the individuals that were newly diagnosed with HIV-1 infection among IDUs living in Guangzhou, between 2008 and 2015. The Pol gene segments of HIV-1 genome from these RNA samples were amplified by nested reverse transcription polymerase chain reaction (Nested-PCR) and sequenced. Subsequently, phylogenetic tree was reconstructed using both pol sequences of samples and references before the subtype of HIV-1 was determined. Distributions of HIV-1 subtypes detected in IDUs with different demographic characteristics in different years were compared. Results: A total of 437 pol gene segments were successfully obtained from 517 serum samples of HIV infection IDUs. The average age of 437 HIV infected IDUs was 37.37 years with standard deviation as 8.17 years. 51.5% (225/437) of the HIV infected IDU that registered residence were not in Guangdong. The Guangxi Registered residents were accounted for 54.2% (122/225). Proportion of subtype CRF07_BC (46.5%) appeared the highest, followed by CRF01_AE (24.3%), CRF08_BC (23.3%) and other subtypes (5.9%). The annual proportions of subtype CRF07_BC (trend χ²=19.703, P=0.006) and CRF08_BC (trend χ²=25.718, P=0.001) were significantly different. The proportion of subtype CRF07_BC decreased from 56.9% to 34.2% (trend χ²=15.139, P=0.000), while the proportion of CRF08_BC increased from 11.8% to 37.0% (trend χ²=22.577, P=0.000). The proportion of CRF08_BC was significantly higher in the HIV infected IDUs with Guangxi residence (Monte Carlo simulation of exact probability P=0.000, 99%CI: 0.000-0.000). Conclusions CRF07_BC, CRF01_ AE and CRF08_BC were the predominant HIV-1 subtypes while multiple subtypes were co-circulated among the HIV infected IDUs in Guangzhou, between 2008 and 2015. Behavioral intervention set for HIV infected IDUs with Guangxi residence should be strengthened in Guangzhou.

Fractures are frequently occurring diseases that endanger human health. Crucial to fracture healing is cartilage formation, which provides a bone-regeneration environment. Cartilage consists of both chondrocytes and extracellular matrix (ECM). The ECM of cartilage includes collagens and various types of proteoglycans (PGs), which play important roles in maintaining primary stability in fracture healing. The PG form of dentin matrix protein 1 (DMP1-PG) is involved in maintaining the health of articular cartilage and bone. Our previous data have shown that DMP1-PG is richly expressed in the cartilaginous calluses of fracture sites. However, the possible significant role of DMP1-PG in chondrogenesis and fracture healing is unknown. To further detect the potential role of DMP1-PG in fracture repair, we established a mouse fracture model by using a glycosylation site mutant DMP1 mouse (S89G-DMP1 mouse). Upon inspection, fewer cartilaginous calluses and down-regulated expression levels of chondrogenesis genes were observed in the fracture sites of S89G-DMP1 mice. Given the deficiency of DMP1-PG, the impaired IL-6/JAK/STAT signaling pathway was observed to affect the chondrogenesis of fracture healing. Overall, these results suggest that DMP1-PG is an indispensable proteoglycan in chondrogenesis during fracture healing.

In the original publication, the label of Fig. 2C should be read as "GFAP/lectin/DAPI" not "DMP1/GFAP/lectin/DAPI".

The blood-brain barrier (BBB) is a tight boundary formed between endothelial cells and astrocytes, which separates and protects brain from most pathogens as well as neural toxins in circulation. However, detailed molecular players involved in formation of BBB are not completely known. Dentin matrix protein 1 (DMP1)-proteoglycan (PG), which is known to be involved in mineralization of bones and dentin, is also expressed in soft tissues including brain with unknown functions. In the present study, we reported that DMP1-PG was expressed in brain astrocytes and enriched in BBB units. The only glycosylation site of DMP1 is serine89 (S89) in the N-terminal domain of the protein in mouse. Mutant mice with DMP1 point mutations changing S89 to glycine (S89G), which completely eradicated glycosylation of the protein, demonstrated severe BBB disruption. Another breed of DMP1 mutant mice, which lacked the C-terminal domain of DMP1, manifested normal BBB function. The polarity of S89G-DMP1 astrocytes was disrupted and cell-cell adhesion was loosened. Through a battery of analyses, we found that DMP1 glycosylation was critically required for astrocyte maturation both in vitro and in vivo. S89G-DMP1 mutant astrocytes failed to express aquaporin 4 and had reduced laminin and ZO1 expression, which resulted in disruption of BBB. Interestingly, overexpression of wild-type DMP1-PG in mouse brain driven by the nestin promoter elevated laminin and ZO1 expression beyond wild type levels and could effectively resisted intravenous mannitol-induced BBB reversible opening. Taken together, our study not only revealed a novel element, i.e., DMP1-PG, that regulated BBB formation, but also assigned a new function to DMP1-PG.
Animals ; Astrocytes ; cytology ; metabolism ; Blood-Brain Barrier ; cytology ; metabolism ; Cells, Cultured ; Extracellular Matrix Proteins ; genetics ; metabolism ; Female ; Glycosylation ; Male ; Mice ; Proteoglycans ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction