An Anterior Cingulate Cortex-to-Midbrain Projection Controls Chronic Itch in Mice / 神经科学通报·英文版

Itch is an unpleasant sensation that provokes the desire to scratch. While acute itch serves as a protective system to warn the body of external irritating agents, chronic itch is a debilitating but poorly-treated clinical disease leading to repetitive scratching and skin lesions. However, the neural mechanisms underlying the pathophysiology of chronic itch remain mysterious. Here, we identified a cell type-dependent role of the anterior cingulate cortex (ACC) in controlling chronic itch-related excessive scratching behaviors in mice. Moreover, we delineated a neural circuit originating from excitatory neurons of the ACC to the ventral tegmental area (VTA) that was critically involved in chronic itch. Furthermore, we demonstrate that the ACC→VTA circuit also selectively modulated histaminergic acute itch. Finally, the ACC neurons were shown to predominantly innervate the non-dopaminergic neurons of the VTA. Taken together, our findings uncover a cortex-midbrain circuit for chronic itch-evoked scratching behaviors and shed novel insights on therapeutic intervention.

Corythucha marmorata affects growth and quality of Artemisia argyi / 中国中药杂志

This study aims to investigate the impact of the invasive pest Corythucha marmorata on the growth and quality of Artemi-sia argyi. The signs of insect damage at the cultivation base of A. argyi in Huanggang, Hubei were observed. The pests were identified based on morphological and molecular evidence. The pest occurrence pattern and damage mechanism were investigated. Electron microscopy, gas chromatography-mass spectrometry(GC-MS), and high performance liquid chromatography(HPLC) were employed to analyze the microstructure, volatile oils, and flavonoid content of the pest-infested leaves. C. marmorata can cause destructive damage to A. argyi. Small decoloring spots appeared on the leaf surface at the initial stage of infestation. As the damage progressed, the spots spread along the leaf veins and aggregated into patches, causing yellowish leaves and even brownish yellow in the severely affected areas. The insect frequently appeared in summer because it thrives in hot dry conditions. After occurrence on the leaves, microscopic examination revealed that the front of the leaves gradually developed decoloring spots, with black oily stains formed by the black excrement attaching to the glandular hairs. The leaf flesh was also severely damaged, and the non-glandular hairs were broken, disor-ganized, and sticky. The content of neochlorogenic acid, cryptochlorogenic acid, isochlorogenic acids A and B, hispidulin, jaceosidin, and eupatilin at the early stage of infestation was significantly higher than that at the middle stage, and the content decreased at the last stage of infestation. The content of eucalyptol, borneol, terpinyl, and caryophyllin decreased in the moderately damaged leaves and increased in the severely damaged leaves. C. marmorata was discovered for the first time on A. argyi leaves in this study, and its prevention and control deserves special attention. The germplasm materials resistant to this pest can be used to breed C. marmorata-resis-tant A. argyi varieties.

Risk factors for neutropenia of late newborns / 中国当代儿科杂志

OBJECTIVE@#To study the risk factors and treatment for neutropenia of late newborns (NLN).@*METHODS@#Related clinical data were collected from the preterm infants and critically ill neonates who were admitted to the neonatal intensive care unit from July 2019 to January 2020. A total of 46 newborns with a blood absolute neutrophil count (ANC) of 37 weeks. There was no significant difference between the two groups in the incidence rates of gestational hypertension, premature rupture of membranes > 18 hours and intrauterine distress, 5-minute Apgar score, the duration of positive pressure ventilation, the incidence rate of early-onset sepsis, and the type of initially used antibiotics (@*CONCLUSIONS@#The risk of NLN increases with the presence of late-onset sepsis and the increase in the duration of antibiotic use. NLN is generally a benign process. G-CSF appears to be safe and effective for NLN with severe disease conditions or severe reduction in ANC.

Effect of Sulfur Fumigation on Content of Chemical Constituents in Panacis Quinquefolii Radix and Preliminary Study on Its Identification Method / 中国实验方剂学杂志

Objective: To elucidate the effect of sulfur fumigation on the content of chemical composition and quality of Panacis Quinquefolii Radix, and to discuss the identification method of sulfur fumigated Panacis Quinquefolii Radix. Method: The content changes of ginsenoside Rb1, Re, Rg1 in Panacis Quinquefolii Radix were determined by HPLC-DAD before and after sulfur fumigation. The content changes of 11 inorganic elements in Panacis Quinquefolii Radix before and after sulfur fumigation were detected by microwave digestion method and inductively coupled plasma emission spectrometry tandem mass spectrometry (ICP-MS). The quality difference of Panacis Quinquefolii Radix before and after sulfur fumigation was qualitatively distinguished combined with the principal component analysis. Result: Three ginsenosides were well separated and showed good linearity with R2 ≥ 0.999.Before sulfur fumigation, the contents of ginsenoside Rg1, Re and Rb1 in Panacis Quinquefolii Radix were (0.897±0.045), (13.092±0.639), (12.896±0.430) g ·kg-1, respectively. After sulfur fumigation, the contents of these three components in Panacis Quinquefolii Radix were (0.807±0.032), (11.968±0.627), (11.752±0.696) g ·kg-1, respectively. After fumigation, the contents of inorganic elements (Cr, Mn, Co, Cu, As, Se, Ag, Cd, Sb, Pb and Fe) from Panacis Quinquefolii Radix increased in varying degrees. According to the distribution results of three-dimensional (3D) scattered points, sulfur and non-sulfur fumigation products of Panacis Quinquefolii Radix could be effectively distinguished. Conclusion: After sulfur fumigation, the contents of ginsenoside Re and Rb1 in Panacis Quinquefolii Radix decrease significantly, but the contents of 11 inorganic elements increase, suggesting that sulfur fumigation may have a great influence on the quality of Panacis Quinquefolii Radix.

DNA methylation of HDAC4 gene affects the trans-differentiation process of hMSCs into sweat gland like cells / 中国应用生理学杂志

OBJECTIVE@#To investigate the changesof DNA methylation in histone deacetylases 4 gene (HDAC4) and its effectduring the trans-differentiation process of human mesenchymal stem cells (hMSCs) into sweat gland like cells (SGLCs).@*METHODS@#Selected cell lines of human mesenchymal stem cells (hMSCs) were cultured and expended , the third generation ofhMSCs and heat-shocked sweat gland cells were picked up, and were co-culturedwith adding inducible factor in the transwell chamber. The sweat gland like cells (SGLCs)in experiment group and the hMSCs in control group were collected, the changes of DNA methylation degree of CpG dinucleotide sitesin histone deacetylases 4 gene (HDAC4) promotor were detected by methylation specific PCR (MSP)andMaldi-TOF Mass Array. And then, the hMSCs in experiment group were treated with 5-aza-CdR (5-aza-2-deoxycytidine, 10 μmol/L), while the hMSCsin control group were culturedwith PBS at the same time. ThemRNA expressions of HDAC4 gene and carcino-embryonic antigen (CEA)gene in the two groups were measured by RT-PCR.@*RESULTS@#The methylation of HDAC4gene in hMSCs was in high level before induction, the methylation degreeof CpG dinucleotide sites located in cg2463009 was 0.901, and the methylation degree of HDAC4gene in SGLCs was markedly decreased by 37% after induction, which was 0.531. The methylationlevel of CpG dinucleotide sites located in cg14823429was changed from 0.687to 0.386 after induction. The mRNA expression of HDAC4 gene was upregulated in test group after treated with 5-aza-CdR for 48 hours, the mRNA expression of CEA gene related with transdifferentiation was enhanced too at the same term, there was significantly statistic difference compared with control group (<0.05).@*CONCLUSIONS@#Methylation of HDAC4 gene participates in the regulation of the trans-differentiation of hMSCs into sweet gland like cells.

Construction of expression vector pLCK-CD69-IRES-EGFP and generation of CD69 transgenic mice / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To construct the expression vector pLCK-CD69-IRES-EGFP that contains mouse cell surface activation protein CD69 and enhanced green fluorescent protein(EGFP),and to generate CD69 transgenic mice based on this vector.</p><p><b>METHODS</b>First, RNA was extracted from mouse lung tissue and cDNA was synthesized via reverse transcription. PCR primer was designed through the PubMed searching, then mouse CD69 DNA fragment was amplified with PCR. Second, this DNA fragment was subcloned to the pInsulater-LCK-IRES-EGFP plasmid and constructed the transgenic vector after the verification of nucleotide sequence. Third, the expression vector was then transfected into 293 T cells and its expression in 293 T cells was observed under fluorescence microscope. Last, microinjection was performed to transfer the expression vector pLCK-CD69-IRES-EGFP into fertilized eggs, which were implanted into pseudo-pregnant recipient mice. After birth the tail samples of the pups were obtained for the purpose of genotyping to determine the transgenic founders. Fluorescence microscope and flow cytometer were used to measure the expression of CD69 on cells.</p><p><b>RESULTS</b>The construction of the expression vector pLCK-CD69-IRES-EGFP was verified by enzyme digestion and DNA sequencing. The transfected 293 T cell showed expression of the protein under fluorescence microscope. Identification of PCR for the tail tissue of the pups confirmed the present of CD69 transgene and resting lymphocytes demonstrated the expression of CD69.</p><p><b>CONCLUSION</b>The construction of expression vector pLCK-CD69-IRES-EGFP and generation of CD69 transgenic mice have been successfully processed, which lays a foundation of the solid pattern studies in inflammatory diseases.</p>

Correlation of Eps8 with proliferation, metastasis and prognosis of malignant tumors / 中国实验血液学杂志

Epidermal growth factor receptor pathway substrate 8 (Eps8) is one of crucial kinase substrates for the epidermal growth factor receptors. Eps8 is related to mitosis and differentiation of normal cells. In recent years, it has been demonstrated that Eps8 involves in proliferation, metastasis and prognosis of many malignant tumors. Experiments have shown that Eps8 involves in Ras-Rac pathway of EGFR signaling by forming Eps8-Abi1-Sos1 tri-complex or participates in endocytosis mediated by rab5. Furthermore, Eps8 has also been found to regulate cell cycle. In conclusion, it may become a monitor and a new target for the treatment of malignant tumors. This review briefly introduces molecular structure and physiological function of Eps8, focusing on its function and molecular mechanism in proliferation, metastasis and prognosis of malignant tumors.

Tripterygium polyglycosid attenuates the established airway inflammation in asthmatic mice / 中国结合医学杂志

<p><b>OBJECTIVE</b>To investigate the effect of Tripterygium polyglycosid on establishing airway eosinophil infiltration and related airway hyperresponsiveness of asthmatic mice.</p><p><b>METHODS</b>A mature murine asthmatic model was made with ovabulmin sensitized and challenged C57BL/6 mice. Forty mice were divided into four groups with 10 mice in each group: mice sensitized and challenged with saline (WS group), mice sensitized and challenged with ovalbumin (WO group), mice sensitized and challenged with ovalbumin and treated with Tripterygium polyglycosid (TP group) and Dexamethasone (DXM group). The mice were intraperitoneally injected with 20 μg chicken ovabulmin emulsified in injected alum on days 0 and 14, then were challenged with an aerosol generated from 1% ovabulmin on days 24, 25 and 26. Tripterygium polyglycosid was injected intraperitoneally at 50 mg/kg on days 25, 26 and 27 after ovabulmin challenge. Dexamethasone was administrated to mice at 2 mg/kg on day 21, 23 before ovabulmin challenge. The airway hyperresponsiveness, mucus production, eosinophils in parabronchial area and bronchoalveolar lavage fluid and the level of interleukin-5, granulo-macrophage clone stimulating factor in bronchoalveolar lavage fluid were measured as indexes of inflammation.</p><p><b>RESULTS</b>Tripterygium polyglycosid treatment after ovabulmin challenge completely inhibited eosinophil infiltration in bronchoalveolar lavage fluid [(0.63 ± 0.34)× 10(4) vs. (75.0 ± 14.8)× 10(4), P<0.05] and the peribrochial area (12.60 ± 3.48 mm(2) vs. 379.0 ± 119.3 mm(2), P<0.05), mucus overproduction in airway (2.8 ± 1.7 vs. 7.1±5.6, P<0.05), and increased interleukin-5 levels in bronchoalveolar lavage fluid (28.8 ± 2.8 pg/mL vs. 7.5 ± 3.5 pg/mL, P<0.05). Meanwhile, Tripterygium polyglycosid treatment after ovabulmin challenge also partially inhibited airway hyperresponsiveness. The level of granulo-macrophage clone stimulating factor in bronchoalveolar lavage fluid didn't change with drugs intervention.</p><p><b>CONCLUSIONS</b>The administration of Tripterygium polyglycosid could inhibit the established airway inflammation and reduce the airway hyperresponsiveness of allergic asthmatic mice. It provides a possible alternative therapeutic for asthma.</p>

Skin irritation and sensitization of swine acellular dermal matrix treated with hyaluronic acid / 中华烧伤杂志

<p><b>OBJECTIVE</b>To evaluate the skin irritation and sensitization potential of the swine acellular dermal matrix treated with hyaluronic acid (SADM-HA).</p><p><b>METHODS</b>(1) Skin irritation test. Twelve New Zealand rabbits were divided into SADM-HA group, allogeneic skin group, and (human) xeno-skin group according to the random number table, with 4 rabbits in each group. Four test sites were designed on the back of each rabbit. Two test sites of each rabbit in the three groups were covered with SADM-HA, allogeneic skin, and xeno-skin, respectively. Another test site was covered with gauze containing 200 g/L sodium dodecyl sulfate solution as positive control. The last test site was covered with gauze containing normal saline as negative control. The primary irritation index and cumulative irritation index of each material were calculated. (2) Skin closed-patch test. Sixty guinea pigs were used. Fifty-four guinea pigs were divided into SADM-HA group, allogeneic skin group, and (human) xeno-skin group according to the random number table, with 18 guinea pigs in each group. Twelve guinea pigs in each of the three groups were correspondingly induced and stimulated by SADM-HA, allogeneic skin, and xeno-skin, with 6 guinea pigs in each group treated with ethanol-soaked gauze to serve as negative control. The remaining 6 guinea pigs were treated with gauze containing 25% α-hexylcinnamaldehyde ethanol solution as positive control. The rating scales of Magnusson and Kligman were used to grade the condition of skin after being treated with above-mentioned materials to evaluate skin sensitivity to them at post stimulation hour 24 and 48. Data were processed with the non-parametric test of independent samples.</p><p><b>RESULTS</b>(1) In the skin irritation test, the primary irritation indexes of the three dressings in SADM-HA group, allogeneic skin group, and xeno-skin group were respectively -0.04, 0.13, and 0.08. The cumulative irritation indexes of the three dressings in SADM-HA group, allogeneic skin group, and xeno-skin group were respectively 0.27, 0.10, and 0.25, which were close to those of negative control within the three groups. The skin irritation of each of the three materials was negligible. (2) In the skin closed-patch test, all scores of the three dressings in SADM-HA group, allogeneic skin group, and xeno-skin group were between 0 and 1. The scores of SADM-HA group and allogeneic skin group were close to those of negative control within the two groups (with U values respectively 188.00 and 90.00, P values both above 0.05). The differences were statistically significant between each material of the three groups and positive control (with U values respectively 19.00, 59.00, 21.50, P values all below 0.01).</p><p><b>CONCLUSIONS</b>The SADM-HA is safe and reliable without skin irritation and sensitization, and it has encouraging prospect in clinical application.</p>

Influence of the number of removed axillary lymph nodes on the prognosis of node-negative primary breast cancer / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To analyze the relationship between the number of removed axillary lymph nodes and prognosis of axillary node-negative breast cancer.</p><p><b>METHODS</b>The clinicopathological data of 655 patients with breast cancer were analyzed retrospectively. The disease-free survival curves were generated according to the number of removed axillary lymph nodes using Kaplan-Meier plots. The correlation between the co-variables and rate of breast cancer-related events was analyzed using Cox model.</p><p><b>RESULTS</b>The overall five year-disease free survival rate of the 655 cases was 94.4%. The rate of patients with lymph node number ≤ 12 was 90.3%, and that of lymph node number > 12 was 96.5%, with a statistically significant difference (P = 0.009). Significantly less breast cancer-related events were observed in patients with lymph node number > 12 (15/426, 3.5%) than that in patients with lymph node number ≤ 12 (22/229, 9.6%) (P = 0.009).</p><p><b>CONCLUSIONS</b>When axillary node dissection is indicated, dissection of lymph nodes >12 leads to much less breast cancer-related events than that in patients with dissected lymph node ≤ 12. The more lymph nodes are dissected, the more accurate prognosis can be estimated.</p>

Retinal morphologic change and related factor analysis in rats with early stage of diabetes mellitus / 中华实验眼科杂志

Background The pathological foundation of diabetic retinopathy is the breakdown of the bloodretina barrier induced by multifactors.Objective This study was to investigate the retinal morphologic change and the levels of serum vascular endothelial growth factor(VEGF),endothelin(ET)and nitric oxide(NO)in diabetic rats.Methods Forty healthy female SD rats were randomly divided into experimental group and control group and 20 rats for each group.Acute diabetes models were established by the intraperitoneal injection of 60 mg/kg streptozotocin in experimental group and the equal volume of buffer solution was injected at the same way in the control group.The serum VEGF level,ET level and NO concentration of diabetic and control rats were detected using ELISA double antibody sandwich method,125I radioimmune method and nitrate reduction method respectively at 2,4,6 and 8 weeks after injection.The eyeballs of rats were enucleated at the eighth week for retinal pathologic examination.This experiment followed the Measures for the administration of experimental animals of Shanghai City.Results The retinal structure was normal throughout the experimental duration in the control group.However,retinal edema and cellular disorganization appeared at 4 months and retinal blooding could be seen with the extending of diabetes course.The levels of serum VEGF and ET in each experimental group were significantly higher than those in control groups(P＜0.05).The levels of serum NO elevated in 2-month experimental group compared with same-phase control group(Z =-2.193,P＜0.05),and those in 6-and 8-month experimental groups were significantly lower than in corresponding control groups(Z =-2.449,Z =-2.236,P＜0.05).With the progression of the disease,the levels of VEGF and ET increased gradually,but the levels of NO decreased gradually,showing statistically significant differences(P＜0.05).The level of serum VEGF showed a positive correlation with serum ET level and a negative correlation with the serum NO concentration(r=-0.814,r=-0.803,P＜0.01)in the experimental group.A negative leaner relation was also found between serum ET level and serum NO concentration(r=0.821,P＜0.01).Conclusions The serum VEGF,ET and NO levels are closely associated with the degree of retinal lesion in early diabetic models.These results suggest that serum VEGF,ET and NO levels may be the important indexes predicting the course of retinal disease in diabetic rats.

CD69 expression on airway eosinophils and airway inflammation in a murine model of asthma / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Asthma is a chronic airway disease with inflammation characterized by physiological changes (airway hyper-responsiveness, AHR) and pathological changes (inflammatory cells infiltration and mucus production). Eosinophils play a key role in the allergic inflammation. But the causative relationship between eosinophils and airway inflammation is hard to prove. One of the reasons is lack of activation marker of murine eosinophils. We investigated the expression of CD69 on murine eosinophils in vitro, the relationship between the expression of CD69 on eosinophils from peripheral blood and bronchoalveolar lavage fluid and on airway inflammation in asthmatic mice.</p><p><b>METHODS</b>Eosinophils from peripheral blood of IL-5 transgenic mice (NJ.1638) were purified. Mice were divided into five groups: wild type mice sensitized and challenged with saline (WS group), wild type mice sensitized and challenged with ovalbumin (WO group), IL-5(-/-) mice sensitized and challenged with saline and transferred with purified eosinophils (ISE group), IL-5(-/-) mice sensitized and challenged with OVA and transferred with purified eosinophils (IOE group), IL-5(-/-) mice sensitized and challenged with OVA and transferred with purified eosinophils, pretreated with anti CD4 monoclonal antibody (IOE+antiCD4mAb group). IL-5(-/-) mice were sensitized with OVA at day 0 and day 14, then challenged with OVA aerosol. On days 24, 25, 26 and 27 purified eosinophils were transferred intratracheally to IL-5(-/-) mice. On day 28, blood and BALF were collected and CD69 expression on eosinophils measured by flowcytometry.</p><p><b>RESULTS</b>Purified eosinophils did not express CD69. But eosinophils cultured with PMA + MA, IFN-gamma, IL-5 or GM-CSF expressed CD69 strongly. Eosinophils from blood of WO, WS group did not express CD69 at all. The numbers of eosinophils in BALF of WO group, IOE group, ISE group and IOE + antiCD4mAb group were significantly higher than in mice of WS group which did not have eosinophils at all. CD69 expression on eosinophils in BALF of IOE and WO groups was strong. Eosinophils in BALF of ISE and IOE + antiCDmAb groups did not express CD69. The mucus production result was similar to CD69 expression. There were eosinophils infiltration in lung slides of all groups except WS group.</p><p><b>CONCLUSION</b>Activation in airway of eosinophils could directly lead to airway inflammation.</p>