Diabetic neuropathic pain (DNP) is the most common disabling complication of diabetes. Emerging evidence has linked the pathogenesis of DNP to the aberrant sprouting of sensory axons into the epidermal area; however, the underlying molecular events remain poorly understood. Here we found that an axon guidance molecule, Netrin-3 (Ntn-3), was expressed in the sensory neurons of mouse dorsal root ganglia (DRGs), and downregulation of Ntn-3 expression was highly correlated with the severity of DNP in a diabetic mouse model. Genetic ablation of Ntn-3 increased the intra-epidermal sprouting of sensory axons and worsened the DNP in diabetic mice. In contrast, the elevation of Ntn-3 levels in DRGs significantly inhibited the intra-epidermal axon sprouting and alleviated DNP in diabetic mice. In conclusion, our studies identified Ntn-3 as an important regulator of DNP pathogenesis by gating the aberrant sprouting of sensory axons, indicating that Ntn-3 is a potential druggable target for DNP treatment.
Mice ; Animals ; Diabetes Mellitus, Experimental/metabolism* ; Axons/physiology* ; Diabetic Neuropathies ; Sensory Receptor Cells/metabolism* ; Neuralgia/metabolism*

Objective: To observe the association between puberty development and thyroid function among school-aged girls in Minhang District of Shanghai, in order to explore the effect of puberty development on thyroid function. Methods: The study was based on a cohort of adolescent girls recruited in iodine-suitable areas of Minhang District, and the baseline and follow-up survey have been carried out from January to March 2019. The method of phased cluster sampling was used to select one junior high school in the east, south, north and middle of Minhang District, Shanghai, respectively. Finally, 464 new junior high school girls were included in the Cohort study for physical examination, and girls were followed up from January to March 2021. The Puberty Development Scale (PDS) was used to assess the stage of puberty. Serum thyroid stimulating hormone (TSH), free triiodothyronine (FT3), and free thyroxine (FT4) were determined for each participant. Thyroid homeostasis structure parameters (THSPs) was calculated. Changes of Thyroid hormones at baseline and follow-up were compared by Wilcoxon signed rank sum test. Multiple linear regression analyses were used to evaluate the associations of thyroid hormones (THs) and THSPs changes with pubertal develepment. Results: Serum TSH levels of female studentds decreased significantly, while their FT3 and FT4 levels increased significantly during the study period ( Z=-10.53, -4.71, -12.46, P <0.01). In multiple linear regression analysis after adjustment for co-variables (including baseline age, change of BMI and waist circumference), FT4 and thyroid feedback quantile-based index (TFQI) in the higher puberty category scores changes (△PCS) group were further reduced compared with those in the low △PCS group ( β =-0.66, -0.55 ). Compared to the late puberty at baseline and follow-up (BLFL) group, FT4 and TFQI showed higher decline in the pre-puberty at baseline and late puberty at follow-up (BPFL) group with the pre-puberty at baseline and end of puberty at follow-up (BPFT) group ( β =-0.55, -0.44)( P <0.05). There were no association of △TSH, △FT3, △FT4/FT3 and TSH index changes (△TSHI) with △PCS or the puberty pattern. Conclusion Serum TSH decreases while serum FT3 and FT4 increase among girls during puberty. Both the initial stage and the velocity of pubertal development are related to thyroid hormone fluctuations.

Nerve regeneration in adult mammalian spinal cord is poor because of the lack of intrinsic regeneration of neurons and extrinsic factors - the glial scar is triggered by injury and inhibits or promotes regeneration. Recent technological advances in spatial transcriptomics (ST) provide a unique opportunity to decipher most genes systematically throughout scar formation, which remains poorly understood. Here, we first constructed the tissue-wide gene expression patterns of mouse spinal cords over the course of scar formation using ST after spinal cord injury from 32 samples. Locally, we profiled gene expression gradients from the leading edge to the core of the scar areas to further understand the scar microenvironment, such as neurotransmitter disorders, activation of the pro-inflammatory response, neurotoxic saturated lipids, angiogenesis, obstructed axon extension, and extracellular structure re-organization. In addition, we described 21 cell transcriptional states during scar formation and delineated the origins, functional diversity, and possible trajectories of subpopulations of fibroblasts, glia, and immune cells. Specifically, we found some regulators in special cell types, such as Thbs1 and Col1a2 in macrophages, CD36 and Postn in fibroblasts, Plxnb2 and Nxpe3 in microglia, Clu in astrocytes, and CD74 in oligodendrocytes. Furthermore, salvianolic acid B, a blood-brain barrier permeation and CD36 inhibitor, was administered after surgery and found to remedy fibrosis. Subsequently, we described the extent of the scar boundary and profiled the bidirectional ligand-receptor interactions at the neighboring cluster boundary, contributing to maintain scar architecture during gliosis and fibrosis, and found that GPR37L1_PSAP, and GPR37_PSAP were the most significant gene-pairs among microglia, fibroblasts, and astrocytes. Last, we quantified the fraction of scar-resident cells and proposed four possible phases of scar formation: macrophage infiltration, proliferation and differentiation of scar-resident cells, scar emergence, and scar stationary. Together, these profiles delineated the spatial heterogeneity of the scar, confirmed the previous concepts about scar architecture, provided some new clues for scar formation, and served as a valuable resource for the treatment of central nervous system injury.
Mice ; Animals ; Gliosis/pathology* ; Cicatrix/pathology* ; Spinal Cord Injuries ; Astrocytes/metabolism* ; Spinal Cord/pathology* ; Fibrosis ; Mammals ; Receptors, G-Protein-Coupled

Objective To investigate the expression and clinical significance of TM9SF3 in lung adenocarcinoma (LUAD). Methods TCGA and GEPIA databases were used to screen the differentially-expressed TM9SF family molecules and analyze their effects on patient prognosis with LUAD. The expression and localization of TM9SF3 in LUAD patients were verified by a human proteomic mapping database, Western blot assay, and polymerase chain reaction assay. Herein, the GSEA was used for the signal pathway enrichment analysis of TM9SF3-related genes. Meanwhile, the TIMER database and CIBERSORT algorithm were used to analyze the correlation between differentially-expressed TM9SF3 and the degree of immune cell infiltration. Results The expression of TM9SF3 in LUAD was significantly increased and had a significant adverse effect on the prognosis of LUAD patients. In addition, immunoblotting and polymerase chain reaction confirmed that TM9SF3 was highly expressed in LUAD. Meanwhile, the genes related to TM9SF3 expression were mainly enriched in cell signaling pathways regulating immune cell activity. The expression of TM9SF3 was significantly correlated with the expression changes of six immune cells. Conclusion TM9SF3 is differentially expressed in LUAD and may be used as a potential prognostic marker for LUAD patients. TM9SF3 can also change the level of immune cell infiltration in LUAD patients and is expected to be a new potential target for LUAD immunotherapy.

ObjectiveThis study aimed to understand the epidemiological characteristics of hand-foot-mouth disease （HFMD） in Minhang District， Shanghai from 2009 to 2020， and provide a scientific basis for the prevention and control of HFMD. MethodsThe case information of HFMD was collected from the National Notifiable Infectious Diseases Reporting System of Chinese Center for Disease Control and Prevention. We used descriptive epidemiological methods to analyze the population characteristics， temporal and spatial distribution of HFMD， the pathogen composition of the case and its changing trend. ResultsFrom 2009 to 2020， a total of 66，198 cases of HFMD were reported in Minhang District， Shanghai， including 377 severe cases （severe case rate 0.57%） and 3 deaths （severs case fatality rate 0.80%）. There were more cases of HFMD in boys than in girls （1.5∶1）. HFMD patients aged under 5 years predominated， accounting for 88.91% of all cases. Majority of the cases （91.42%） were in scattered children （55.80%） and children in kindergartens （35.62%）. The incidence showed a cyclical trend， with low incidence years and high incidence years appearing alternately. The peak period was from April to July， and sometimes there were small peaks during October to December. A total of 12 years time-space scanning analysis revealed 3 clusters. The cluster centers were located in Wujing Town， Huacao Town and Xinzhuang Town， respectively. The proportion of EV71 in common cases was generally decreasing， and reduced to zero in 2019. The proportion of CoxA6 had increased year by year， and reached 75.00% in 2020. CoxA6 became the dominant pathogen in recent years. The number of severe cases had decreased year by year since 2010， and the dominant pathogen was EV71 （90.03% on average） in severe cases. ConclusionThe incidence of HFMD in Minhang District of Shanghai has a downward trend from 2014. The dominant pathogen changes from EV71 to CoxA6， and the dominant pathogen in severe cases is EV71. The discovered temporal and spatial clustering pattern is helpful for in-depth understanding of the distribution and epidemic trend of HFMD in Minhang District， and provides a scientific basis for epidemic prevention and control.

Objective To investigate the correlation between smoking, white blood cell counts, and serum bilirubin levels in male radiation workers, and to explore the role of systemic inflammatory response in the changes of serum bilirubin level induced by smoking. Methods Occupational health examination data of 1 320 male radiation workers in a medical institution was collected. Linear regression analysis method was used to analyze the correlations between smoking and white blood cell counts, between smoking and serum bilirubin levels, and between white blood cell counts and serum bilirubin levels. Results There was a significant positive correlation between smoking and white blood cell counts (P< 0.05). Both smoking and white blood cell counts were significantly negatively correlated with serum bilirubin levels (P < 0.05). However, after adjusting the white blood cell counts, the correlation between smoking and serum bilirubin levels was not statistically significant (P > 0.05). By dividing white blood cell counts into two groups, an inverse correlation was found between smoking and serum bilirubin levels in the high white blood cell count group (P < 0.05). Conclusion Serum bilirubin may be an effective indicator of early health damage caused by smoking in male radiation workers. Smoking may induce inflammatory reaction, thus deplete serum bilirubin and cause its levels to drop.

Objective: To investigate thyroid hormone concentration and associated factors among pubertal girls in Minhang District of Shanghai. Methods: From January to March 2019, a stratified sampling method was used to select junior high schools from the east, south, north, and middle areas in Minhang district. A total of 386 girls of grade 6 in selected schools were included in the study. Physical examination was conducted, and their urine and blood samples were collected to determine urinary iodine concentration (UIC) and thyroid function. Puberty Development Self-rating Scale (PDS) was used to define the pubertal stage. Logistic regression models were conducted to analyze the associations between pubertal stage and thyroid function. Results: The median urinary iodine concentration was 163.57(106.57, 232.96) μg/L. The geometric mean values of TSH, TT3, FT3 and FT4 were 0.29 mU/L, 0.26 nmol/L, 0.68 pmol/L and 1.18 pmol/L.The mean value of TT4 was 91.64 nmol / L. The abnormal rates of TGAb and TPOAb were 6.22% and 4.15%. The rate of abnormal TGAb combined with abnormal TPOAb was 3.68%. Girls in puberty and post-puberty had the lower level of TT4 (OR=0.47,0.43) as compared with girls in pre-puberty stage. Obese girls had higher level of TT3(OR=9.08, 95%CI=1.52-54.07). With the increase of exercise time(0.5-1, >1 h/d), FT4 level was increased (OR=2.45, 2.19). TSH levels were significantly higher in girls with higher TGAb and TPOAb. Girls had higher TT4 or FT4 levels if their TGAb levels were higher and TPOAb levels were normal. Conclusion There is an association between pubertal stage, obesity, exercise and thyroid function in school-aged girls during puberty in iodine sufficient areas. TSH, FT4 and TT4 levels are correlated with TGAb and TPOAb levels.

Objective To find small molecules binding specifically to signal transducer and activator of transcription3 (STAT3) based on surface plasmon resonance (SPR) technology and confirm their inhibitory activities to STAT3. Methods The biomolecular interaction analysis T200 system based on SPR technology was used to couple the purified protein STAT3 to CM5 chip under the optimal pH conditions. The compounds with high binding response value were screened out from 50 candidate compounds derived from traditional Chinese medicines and the binding specificity was then confirmed. Biological experiments were performed to confirm the inhibitory effects of the screened compounds on STAT3. The binding pattern of STAT3 and the compound was fitted by molecular docking technique. Results More than 10 candidate molecules exhibited binding activities to STAT3 and kinetics assays revealed that only one candidate molecule, apigenin, showed specific binding. Western-blot analysis exhibited that apigenin inhibited the phosphorylation of STAT3 dose-dependently. Luciferase reporter gene assays demonstrated that apigenin also inhibited IL-6-induced STAT3 transcriptional activity in a dose-dependent manner. Molecular docking results showed that apigenin binds to the SH2 domain of STAT3, and interacts with key residues Glu638, Gln644, Gly656 and Lys658 by hydrogen bonds and with Tyr657 through π-π interactions. Conclusion Apigenin was a direct inhibitor of STAT3.

OBJECTIVE：To establish the quality standard of the Tibetan medicine Ma lus Fructus （known as the fruit of Ese ）， and to provide scientific reference for quality control and utilization of resource. METHODS ：The qualitative analysis of 20 batches of Malus Fructus with different producing areas and different origins was carried out by morphological observation ，microscopic and TLC identification. According to the method stated in 2015 edition of Chinese Pharmacopoeia ，water content ，total ash content and extract content were determined. HPLC method was used to determine the contents of 6 kinds of flavones （chlorogenic acid ， hyperin，isoquercitrin，quercitin，phloridzin，quercetin）in Malus Fructus. RESULTS ：The properties of this medicinal material were obovate ，spheroidal or oval ，its surface was reddish brown to dark red ，with slight shrinkage. Its taste was sweet ，sour and slightly astringent. The microscopic characteristics of the powder were clear ，the stone cells were scattered individually or clustered，and the pulp parenchyma cells contained calcium oxalate cube crystal and cluster crystal. The results of TLC showed that there were fluorescent spots of the same color in the chromatogram of the tested product and the corresponding position of the reference substance （ursolic acid ）. The content ranges of moisture ，total ash and extracts were 9.02％-11.39％，2.29％-3.35％， 37.12％-58.41％ respectively. The linear ranges of chlorogenic acid ，hyperin，isoquercitrin，，quercetin phlorizin and quercetin were 3.12-50，1.25-20，1.25-20，3.12-50，18.7-300，0.5-8 μg/mL（all r were not less than 0.999 7）. RSDs of prescision ，stability and reproducibility tests were all lower than 3.0％（n＝6）. The average recoveries were 97.56％，98.28％，106.14％，103.74％， 98.37％，98.76％；RSDs were 1.57％，1.62％，1.38％，1.99％，2.15％，2.31％（n＝6）. The content ranges of the above six components in 20 baches of samples were 0.001 5％-0.028 8％，0.005 9％-0.022 0％，0.008 7％-0.030 4％，0.019 4％- 0.048 5％，0.068 5％-0.199 8％ and 0.005 1％-0.081 4％. CONCLUSIONS ：Established method is accurate and stable ，and can be used for quality control of Malus Fructus. It is preliminarily Δ 基金项目：四川省科技计划项目（No.2020YFS0495）；“杏林学 proposed that moisture and total ash content should not exceed 者”学科人才科研提升计划（No.CXTD2018016） 12％ and 3.5％，and extract （by dry product ）and phlorizin ＊硕士研究生 。研究方向 ：中药品种 、质量与资源 。E-mail： content should not be less than 40％ and 0.1％ respectively. 1159020313@qq.com # 通信作者：教授。研究方向：中药品种、质量与资源。E-mail：

A sensitive, rapid, simple and economical ultra-performance liquid chromatography–tandem mass spectro-metric method (UPLC–MS/MS) was developed and validated for simultaneous determination of imatinib, dasatinib and nilotinib in human plasma using gliquidone as internal standard (IS). Liquid-liquid extraction method with ethyl acetate was used for sample pre-treatment. The separation was performed on an Xtimate Phenyl column using isocratic mobile phase consisting of A (aqueous phase: 0.15% formic acid and 0.05% ammonium acetate)and B(organic phase:acetonitrile)(A:B=40:60,v/v).The flow rate was 0.25 mL/min and the total run time was 6 min. The multiple reaction monitoring (MRM) transitions, m/z 494.5→394.5 for imatinib, 488.7→401.5 for dasatinib, 530.7→289.5 for nilotinib and 528.5→403.4 for IS, were chosen to achieve high selectivity in the simultaneous analyses. The method exhibited great improvement in sensitivity and good linearity over the concentration range of 2.6–5250.0 ng/mL for imatinib, 2.0–490.0 ng/mL for dasatinib,and 2.4–4700.0 ng/mL for nilotinib.The method showed acceptable results on sensitivity,specificity, recovery, precision, accuracy and stability tests. This UPLC–MS/MS assay was successfully used for human plasma samples analysis and no significant differences were found in imatinib steady-state trough concentra-tions among the SLC22A5?1889T>C or SLCO1B3 699G>A genotypes(P>0.05).This validated method can provide support for clinical therapeutic drug monitoring and pharmacokinetic investigations of these three tyrosine kinase inhibitors(TKIs).