Objective To investigate the myelination of rat brain at different developmental stages.Methods Luxol fast blue staining,immunohistochemistry and Western blotting were applied to determine the distribution and alternation of myelin sheath in the brains of SD rats at different developmental stages.Results Negative LFB and MBP staining were shown at the stages of embryonicage 18 d(E18),postnatal age 0 d(P0) and P2.At P7,the corpus callosum was weakly stained by LFB.The positive stain of LFB and MBP occurred in P15 on the corpus callosum.With the development going on,the stain turned out stronger,especially in P30,which was similar with that in P90 and P720.The results of Western blot analysis showed the expression of MBP in rat brain was gradually increased with the development of rat brain.Conclusion The myelination starts before P15 in rat brain and becomes mature in P30.The optimal observation time of myelin is 30 d born after.

Objective To evaluate the in vitro effect of ferulic acid on the proliferation of,as well as melanin synthesis,tyrosinase activity and expressions of c-kit and ERK proteins in cultured normal human epidermal melanocytes.Methods Cultured normal human epidermal melanocytes were treated with various concentrations of ferulic acid for different durations,and those remaining untreated served as the control.Then,3-(4,5-dimethylthiazol2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium,inner salt (MTS) assay was performed to estimate cell proliferative activity at 24,48 and 72 hours,sodium hydroxide solubilization method to quantify melanin content in melanocytes at 72 hours,dopa oxidation assay to evaluate tyrosinase activity at 72 hours,Western blot to measure the expressions of c-kit and ERK1/2 proteins at 72 hours.Results Cellular proliferative activity was significantly inhibited in melanocytes treated with ferulic acid of 0.01,0.1 and 1 mg/ml for 24,48 and 72 hours compared with untreated melanocytes (all P ＜ 0.05),and the 72-hour treatment with ferulic acid of 1 mg/ml showed the strongest inhibitory effect.Ferulic acid at 0.01,0.1 and 1 mg/ml all markedly suppressed melanin synthesis and tyrosinase activity,decreased the expressions of c-kit and ERK1/2 proteins in melanocytes,with significant differences in these parameters between ferulic acid-treated and untreated melanocytes (all P ＜ 0.05).Conclusions Ferulic acid could downregulate the proliferation of,as well as melanin synthesis,tyrosinase activity,and expressions of c-kit and ERK proteins in cultured human epidermal melanocytes.

Objective To evaluate the improvement effect of Nerve Growth Decoction ( NGD) on symptoms in senile Parkinson's disease (PD) patients with multi-scales.Methods Totally 100 PD patients who have previously taken Madopa therapy were divided into NGD treatment group(70 cases) and control group( 30 cases) .The NGD treatment group took a package of NGD oral solution twice a day for 30 days,and went on next period of 30 days after an interval of 1~2 weeks.The Unified Parkinson disease Rating Scale (UPDRSⅡ,UPDRSⅢ),the PD NMS Questionnaire (NMSQuest),Activity of Daily Living Scale (ADL), the 39-item Parkinson's Disease Questionnaire ( PDQ-39 ) and Schwab & England Daily Living Scale ( Schwab) were used to evaluate both NGD treatment and control group,and within NGD treatment group be-fore and after NGD treatment.Results ( 1) There were significant differences comparing NGD treatment group with control group in ADL,Schwab,PDQ-39 and NMSQuest ( P<0.05) ((27.78±10.54)points,(0.87 ±0.14)points,(26.07±19.18)points,(9.95±4.70)points vs ((34.15±13.88)points,(0.77±0.21)points, (37.47±24.05)points,(13.46±4.01)points,respectively).(2)There were significant differences in UPDRSⅡ,UPDRSⅢ,NMSQuest,ADL,PDQ-39 and Schwab (10.68±7.15)points,(23.79±16.64)points,(33.96± 14.06)points,(0.77±0.21)points,(36.96±24.26)points,(11.96±5.17)points vs ((8.50±6.40)points, (16.79±13.48)points,(27.78±10.54)points,(0.87±0.14)points,(26.07±19.18)points,(9.95±4.70) points,respectively) between before and after treatment in NGD treatment group (P<0.05, P<0.01) .Con-clusion Based on Madopa therapy,the NGD may obviously improves the symptoms,the patient's quality of life and ability of daily life in senile PD patients.Multi-scale evaluations can be used systematically to evalu-ate PD patients,and as assessment indicators for effects of Traditional Chinese drugs.

To explore the diagnostic value of 75 commonly used clinical laboratory markers for differentiation of traditional Chinese medicine syndromes such as liver and gallbladder damp-heat and liver depression and spleen deficiency in patients with chronic hepatitis B.

Objective To evaluate the relationship between diabetic nephropathy(DN)and diabetic retinopathy(DR)in patients with type 2 diabetes mellitus(T2DM)based on imaging and clinical testing data.Methods Totally 600 T2DM patients who visited the First People's Hospital of Ziyang from March 2021 to December 2022 were included.The fundus photography and fundus fluorescein angiography were performed on all these patients and their age,gender,T2DM duration,cardiovascular diseases,cerebrovascular disease,hypertension,smoking history,drinking history,body mass in-dex,systolic blood pressure,diastolic blood pressure and other clinical data were collected.The levels of fasting blood glu-cose(FPG),triglyceride(TG),total cholesterol(TC),high-density lipoprotein cholesterol(HDL-C),low-density lipo-protein cholesterol(LDL-C),glycosylated hemoglobin(HbA1c),24 h urinary albumin(UAlb),urinary albumin to creati-nine ratio(ACR),serum creatinine(Scr)and blood urea nitrogen(BUN)were measured.Logistic regression was used to analyze the risk factors associated with DR.DR staging was performed according to fundus images,and the convolutional neural network(CNN)algorithm was used as an image analysis method to explore the correlation between DR and DN based on emission computed tomography(ECT)and clinical testing data.Results The average lesion area rates of DR and DN detected by the CNN in the non-DR,mild-non-proliferative DR(NPDR),moderate-NPDR,severe-NPDR and pro-liferative DR(PDR)groups were higher than those obtained by the traditional algorithm(TCM).As DR worsened,the Scr,BUN,24 h UAlb and ACR gradually increased.Besides,the incidence of DN in the non-DR,mild-NPDR,moderate-NPDR,severe-NPDR and PDR groups was 1.67％,8.83％,16.16％,22.16％and 30.83％,respectively.Logistic regression analysis showed that the duration of T2DM,smoking history,HbA1c,TC,TG,HDL-C,LDL-C,24 h UAlb,Scr,BUN,ACR and glomerular filtration rate(GFR)were independent risk factors for DR.Renal dynamic ECT analysis demonstrated that with the aggravation of DR,renal blood flow perfusion gradually decreased,resulting in diminished renal filtration.Conclusion The application of CCN in the early stage DR and DN image analysis of T2DM patients will improve the diag-nosis accuracy of DR and DN lesion area.The DN is worsening as the aggravation of DR.

Objective:To screen important genes and characterize their functions and signaling pathways by weighted gene co-expression network analysis (WGCNA) on the gene expression profile of brain tissue after traumatic brain injury (TBI) so as to provide a reference for the mechanism research and treatment of TBI.Methods:The rat TBI gene expression profile GSE2871 was downloaded from the Gene Expression Omnibus (GEO). The expression profile of 8 799 genes of all 47 rat brain tissue samples was analyzed by WGCNA. After calculating and selecting the β-weighted soft threshold, undirected weighted gene network was constructed to identify gene sets with a high degree of correlation. Sample information was obtained from the database to calculate the correlation between each trait of the samples and modules. Gene ontology (GO) analysis and KEGG pathway analysis were performed for the genes in modules related to injury severity and sampling side in order to unvail the biological processes and pathways involved. The gene-module correlation and gene-trait correlation in these key modules were calculated and hub genes were selected.Results:All the rat brain tissue samples and genes in GSE2871 were included in the WGCNA analysis. A total of 22 modules were obtained, which were marked as modules A to V. Modules E, G, T and U were significantly associated with the sampling side. Modules E and G were significantly related to injury severity . GO analysis and KEGG pathway analysis indicated that the genes in modules E and G with significant relation to injury severity and sampling side were mainly implicated in leukocyte migration, cell chemotaxis, various immune response regulation, etc. The involved pathways included antigen processing and presentation pathways, cell factor-cytokine receptor interaction, interleukin-17 signaling pathway, etc. While modules T and U with significant relation to the sampling side were mainly implicated in hypoxia response, cell metabolism, cell membrane ion channel regulation, signal transduction, etc. The pathways involved were neurodegenerative disease signaling pathways, ribosomes, autophagy, neuroactive ligand-receptor interactions, etc. Among the key modules significantly relating to traits, Tuba1b/1c, Ifitm3, Cebpd, Nfkbia, Serinc3, Pmpcb and Cyp4a8 were selected as hub genes of the above key modules.Conclusion:The genes significantly relating to rat TBI are mainly involved in pathophysiological links such as immune activation, inflammatory response, abnormal energy metabolism, calcium channel disorders, abnormal autophagy and cell apoptosis.

Objective To prepare a nano drug(PFOB@Lip-MMC)with liposome as the carrier,liquid perfluorooc-tyl bromide(PFOB)as core and mitomycin C(MMC)loading on the liposome shell and study its inhibitory effect on the proliferation of human pterygium fibroblasts(HPFs).Methods The thin film dispersion-hydration ultrasonic method was used to prepare PFOB@Lip-MMC and detect its physical and chemical properties.Cell Counting Kit-8,Cam-PI cell viability staining and flow cytometry were employed to detect the impact of different concentrations of PFOB@Lip-MMC on the via-bility of HPFs.DiI fluorescence labeled PFOB@Lip-MMC was used to observe the permeability of the nano drug to HPFs under a laser confocal microscope.After establishing HPF inflammatory cell models,they were divided into the control group(with sterile phosphate-buffered saline solution added),PFOB@Lip group(with PFOB@Lip added),MMC group(with MMC added),PFOB@Lip-MMC group(with PFOB@Lip-MMC added)and normal group(with fresh culture medi-um added)according to the experimental requirements.After co-incubation for 24 h,flow cytometer was used to detect the apoptosis rate of inflammatory cells,and the gene expression levels of interleukin(IL)-1β,prostaglandin E2(PGE2),tumor necrosis factor(TNF)-α and vascular endothelial growth factor(VEGF)in cells were analyzed by PCR.Results The average particle size and Zeta potential of PFOB@Lip-MMC were(103.45±2.17)nm and(27.34±1.03)mV,respec-tively,and its entrapped efficiency and drug loading rate were(72.85±3.28)％and(34.27±2.04)％,respectively.The sustained-release MMC of drug-loaded nanospheres reached(78.34±2.92)％in vitro in a 24-hour ocular surface environ-ment.The biological safety of PFOB@Lip-MMC significantly improved compared to MMC.In terms of the DiI fluorescence labeled PFOB@Lip-MMC,after co-incubation with inflammatory HPFs for 2 h,DiI fluorescence labeling was diffusely dis-tributed in the cytoplasm of inflammatory HPFs.The apoptosis rate of inflammatory HPFs in the PFOB@Lip-MMC group[(77.23±4.93)％]was significantly higher than that in the MMC group[(51.62±3.28)％].The PCR examination results showed that the gene transcription levels of IL-1 β,PGE2,TNF-α and VEGF in other groups were significantly reduced com-pared to the control group and PFOB@Lip group,with the most significant decrease in the PFOB@Lip-MMC group(all P＜0.05).Conclusion In this study,a novel nano drug(PFOB@LIP-MMC)that inhibited the proliferation of HPFs was successfully synthesized,and its cytotoxicity was significantly reduced compared to the original drugs.It has good bio-compatibility and anti-inflammatory effects,providing a new treatment approach for reducing the recurrence rate after pte-rygium surgery.

The vegetative state is a complex condition with unclear mechanisms and limited diagnostic, prognostic, and therapeutic methods. In this study, we aimed to explore the proteomic profile of tears from patients in a traumatic vegetative state and identify potential diagnostic markers using tears-a body fluid that can be collected non-invasively. Using iTRAQ quantitative proteomic technology, in the discovery phase, tear samples collected from 16 patients in a traumatic vegetative state and 16 normal individuals were analyzed. Among 1080 identified tear proteins, 57 were upregulated and 15 were downregulated in the patients compared to the controls. Bioinformatics analysis revealed that the differentially-expressed proteins were mainly involved in the wound response and immune response signaling pathways. Furthermore, we verified the levels of 7 differentially-expressed proteins in tears from 50 traumatic vegetative state patients and 50 normal controls (including the samples used in the discovery phase) using ELISA. The results showed that this 7-protein panel had a high discrimination ability for traumatic vegetative state (area under the curve = 0.999). In summary, the altered tear proteomic profile identified in this study provides a basis for potential tear protein markers for diagnosis and prognosis of the traumatic vegetative state and also provides novel insights into the mechanisms of traumatic vegetative state.
Adult ; Aged ; Aged, 80 and over ; Biomarkers ; metabolism ; Chromatography, Liquid ; Enzyme-Linked Immunosorbent Assay ; Eye Proteins ; metabolism ; Female ; Humans ; Male ; Mass Spectrometry ; Middle Aged ; Persistent Vegetative State ; metabolism ; Proteome ; Proteomics ; ROC Curve ; Tears ; metabolism ; Young Adult

Prognostication of coma patients after brain injury is important, yet challenging. In this study, we evaluated the predictive value of amplitude-integrated electroencephalography (aEEG) for neurological outcomes in coma patients. From January 2013 to January 2016, 128 coma patients after acute brain injury were prospectively enrolled and monitored with aEEG. The 6-month neurological outcome was evaluated using the Cerebral Performance Category Scale. aEEG monitoring commenced at a median of 7.5 days after coma onset. Continuous normal voltage predicted a good 6-month neurological outcome with a sensitivity of 93.6% and specificity of 85.2%. In contrast, continuous extremely low voltage, burst-suppression, or a flat tracing was correlated with poor 6-month neurological outcome with a sensitivity of 76.5% and specificity of 100%. In conclusion, aEEG is a promising predictor of 6-month neurological outcome for coma patients after acute brain injury.
Brain ; physiopathology ; Brain Injuries ; complications ; diagnosis ; physiopathology ; Coma ; diagnosis ; etiology ; physiopathology ; Electroencephalography ; methods ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Neurophysiological Monitoring ; methods ; Prognosis ; Prospective Studies ; Sensitivity and Specificity ; Severity of Illness Index

AIM: To prepare a nanodrug MMC-ATS-@PLGA using polylactic acid hydroxyacetic acid copolymer(PLGA)as a carrier and mitomycin C(MMC)loaded on PLGA, and to analyse the biological safety and treatment effect of this nanodrug on inhibiting the proliferation of filtering bleb scarring after glaucoma surgery in vivo.METHODS: The thin-film dispersion hydration ultrasonic method was used to prepare the MMC-ATS-@PLGA, and its physical and chemical properties were detected. The effect of MMC-ATS@PLGA on rabbit corneas was analysed through corneal fluorescence staining and HE staining, and tear film rupture time(BUT), Schirmer test and intraocular pressure data were collected to analyse ocular surface biosafety. A slit lamp was used to observe and calculate the filtration bubble size, and the tissue morphological changes were analysed by conjunctival HE staining. In addition, immunohistochemistry and Elisa were used to compare the anti-inflammatory effects of Flumiolone Eye Drops(FML), MMC, and MMC-ATS-@PLGA nanoparticles on inhibiting the formation of filtering bleb scarring after glaucoma surgery from multiple perspectives via comparative proteomic analysis.RESULTS: The average particle size and zeta potential of MMC-ATS-@PLGA were 128.78±2.54 nm and 36.49±4.25 mV, respectively, with an encapsulation efficiency and a drug loading rate of(78.49±2.75)% and(30.86±1.84)%, respectively. At 33°C(the ocular surface temperature), the cumulative release rate of the MMC-ATS-@PLGA nanoparticles reached(76.58±2.68)% after 600 min. Moreover, corneal fluorescence staining, HE, BUT, Schirmer, and intraocular pressure results showed that MMC-ATS-@PLGA had good biocompatibility with the ocular surface of rabbits. At 3 wk after surgery, the area of filtering blebs in the MMC-ATS-@PLGA group was significantly larger than that in the FML group and MMC group, and the filtering blebs in the control group had basically disappeared. Pathological tissue analysis of the conjunctiva in the filtering blebs area of the eyes of the rabbits revealed that compared with that in the normal group, the morphology of the collagen fibres in the MMC-ATS-@PLGA group was relatively regular, the fibres were arranged neatly, and the tissue morphology was similar to that of the normal group. Immunohistochemistry and Elisa confirmed that compared with those in the normal group, the expression levels of α-SMA, CTGF, and type Ⅲ collagen fibre antibodies were significantly increased in the control group. After FML, MMC, or MMC-ATS-@PLGA treatment for 3 wk, the expression of inflammatory factors gradually decreased. Among the groups, the MMC-ATS-@PLGA group showed the most significant decrease(P<0.05).CONCLUSION: This study successfully synthesized a nanomedicine(MMC-ATS-@PLGA)that inhibits scar proliferation after glaucoma filtration surgery. The drug had stable physicochemical properties, good biocompatibility, and better anti-inflammatory effects by inhibiting the expression of α-SMA, CTGF, and type Ⅲ collagen fibres, which can prevent the formation of scarring in the filtering blebs area, thereby improving the success rate of glaucoma filtering surgery.