ObjectiveThis paper aims to explore the action and molecular mechanism of modified Tongluo Tangtai Formula（MTLTT） on myelin damage in diabetic peripheral neuropathy （DPN） based on network pharmacology and in vitro experiments. MethodsThe chemical components of the MTLTT were retrieved from the Traditional Chinese Medicine Systems Pharmacology （TCMSP） database and literature， and the component targets were collected from the SwissTargetPrediction database. The targets of DPN were collected from the GeneCards， OMIM， Disgenet， and GEO databases. Gene ontology （GO） and Kyoto encyclopedia of genes and genomes （KEGG） enrichment analyses were performed using the Metascape database， and a network diagram was constructed using Cytoscape software. The binding actions of core components with glycogen synthase kinase 3 beta （GSK-3β） and β-catenin were analyzed by Autodock Vina. An in vitro DPN model was established by high glucose-induced Schwann cells and dorsal root ganglion cells （SCs/DRGs）. The ultrastructural morphological changes of SCs and DRGs were observed by scanning electron microscope（SEM）， and the expressions of myelin-associated glycoprotein （MAG） and myelin basic protein （MBP） were detected by immunofluorescence staining. The mRNA and protein expression levels of MAG， MBP， myelin protein 0 （P0）， peripheral myelin protein 22 （PMP22）， and Wnt/β-catenin signaling pathway-related protein β-catenin， GSK-3β， Wnt family member 3α （Wnt3α）， and Wnt inhibitory factor-1 （Wif-1） were detected by real-time polymerase chain reaction （Real-time PCR） and Western blot. ResultsNetwork pharmacology analysis revealed that MTLTT components may treat DPN via the Wnt signaling pathway， involving key proteins such as GSK-3β， β-catenin and Wif-1. The molecular docking results indicate that atropine， apigenin， baicalein， isoflavanone， and albiflorin have good binding activity with GSK-3β， and that all 13 core components have stable binding activity with β-catenin. Cell experiments showed that compared with the blank group， SCs and DRGs in the model group exhibited severe morphological and structural abnormalities such as disintegration， shrinkage and axonal rupture， while these abnormal changes were improved after MTLTT intervention. Immunofluorescence results indicated that the fluorescence intensity of MAG and MBP was markedly decreased in the model group relative to the blank group（P<0.01）， while MTLTT treatment obviously upregulated the expression of MAG and MBP compared with the model group （P<0.01）. Real-time PCR and Western blot assays revealed that the expression levels of myelin-related molecules MAG， MBP， P0 and PMP22 were significantly reduced in the model group （P<0.05，P<0.01）， and MTLTT remarkably increased their expression levels （P<0.05）. In the Wnt/β-catenin signaling pathway， the mRNA levels of GSK-3β， Wif-1 and Wnt3α were elevated and β-catenin mRNA expression was declined in the model group （P<0.01）. Meanwhile， the protein expressions of GSK-3β and Wif-1 were upregulated， whereas those of Wnt3α and β-catenin were downregulated （P<0.01）. Compared with the model group， MTLTT at different doses reduced the mRNA and protein levels of GSK-3β and Wif-1 to varying degrees （P<0.05）， and distinctly enhanced the protein expression of Wnt3α and β-catenin（P<0.01）. ConclusionMTLTT can alleviate high glucose-induced myelin damage. Its protective mechanism may promote myelin repair by upregulating the expression of MAG， MBP， P0 and PMP22， and the therapeutic effect is possibly associated with the activation of Wnt/β-catenin signaling pathway.

Objective:To explore the genotype and the clinical phenotype of SCN2A-related developmental delay in children. Methods:A case series study was adopted. Collect clinical data from 10 cases of children with SCN2A gene variants diagnosed with global developmental delay/intellectual disability who were admitted to the Children′s Hospital between July 2019 and March 2023. Summarize the clinical phenotype and genotype based on clinical data such as general information, clinical manifestations, imaging examinations, laboratory tests, genetic testing results, and comprehensive pediatric neuropsychological development assessment. Results:A total of 10 patients were recruited, including 7 males and 3 females, with an age range of 27 days to 5 years and 9 months. 9 patients underwent children′s neuropsychological and behavioral assessments, and the results were consistent with global developmental delay, including 2 mild cases, 4 moderate cases, and 3 severe cases. 3 cases had autism spectrum disorder, and 2 cases had epilepsy. 6 patients underwent complete head MRI examination, and 4 of them showed abnormalities, including delayed myelination, widening of the local extra brain space in the frontal lobe, and abnormal frontal lobe morphology. All 10 cases had point variants. Among them, 9 cases are de novo and 1 case is maternal inheritance. Out of 10 cases, there were 5 cases with copy number variations, but all of them were of unknown significance. Among the 10 variants, 8 have been reported and 2 have not been reported, namely c.4145A>T(p.N1382I) and c.4937T>A(p.I1646N). In this study, 4 out of 10 patients with SCN2A variants had variation sites located in the S4 segment of domain which constitute Nav1.2, the sodium ion channel encoded by SCN2A. The developmental quotient level was lower when the variation sites were located in the S4 segment of domain, and the difference was statistically significant ( t=-3.101, P=0.017), indicating that the severity of developmental delay may be related to the localization of amino acids corresponding to variant sites within the protein domain. Conclusion:SCN2A mutations are strongly associated with diverse neurodevelopmental disorders. In this study, the phenotypic spectrum of SCN2A variants encompassed epilepsy, global developmental delay, and autism spectrum disorder. Affected individuals exhibited early-onset developmental delays, predominantly moderate to severe in severity. Voltage-sensing domain dysfunction in sodium channels may constitute a critical pathomechanism underlying neurodevelopmental impairments. Further electrophysiological characterization and molecular mechanistic studies are warranted todelineate the genotype-phenotype correlations between specific variant loci and clinical severity.

Objective: To investigate the effects of cranial electrotherapy stimulation(CES) with varying parameter configurations on sleep quality in patients diagnosed with chronic insomnia disorder. Methods: Seventy-two participants meeting diagnostic criteria for chronic insomnia disorder were randomly allocated to a four-arm parallel study design.The intervention protocol comprised:Group 1(G1) received CES at 0.5 Hz,300 μA;Group 2(G2) underwent CES treatment at 1.5 Hz,300 μA;Group 3(G3) administered 100 Hz,300 μA stimulation;and Group4(G4) received sham stimulation with identical device placement but no current delivery.Primary outcomes were quantified through polysomnography(PSG) recordings conducted at baseline and post-intervention,whereas secondary outcomes were assessed via standardized sleep questionnaires including the Pittsburgh sleep quality index(PSQI) and Insomnia Severity Index(ISI). Results: Following a 10-day intervention protocol,significant clinical improvements were observed across all active treatment groups(G1-G3) as evidenced by reductions in PSQI.Insomnia severity index(ISI) scores quantitative polysomnographic analysis revealed that both G2(1.5 Hz) and G3(100 Hz) cohorts demonstrated statistically significant enhancements in Flinders Fatigue Scale(FFS) scores,total sleep time(TST),and sleep efficiency(SE),accompanied by reduced sleep onset latency(SOL) compared to baseline measurements.However,no statistically significant differences were detected between the G2 and G3 intervention arms across all measured parameters.CES exerted no significant effect on sleep architecture. Conclusion CES can effectively improve the sleep of patients with chronic insomnia.Within a certain range,a higher frequency of CES leads to better sleep improvement effects.

Objective:To explore the genotype and the clinical phenotype of SCN2A-related developmental delay in children. Methods:A case series study was adopted. Collect clinical data from 10 cases of children with SCN2A gene variants diagnosed with global developmental delay/intellectual disability who were admitted to the Children′s Hospital between July 2019 and March 2023. Summarize the clinical phenotype and genotype based on clinical data such as general information, clinical manifestations, imaging examinations, laboratory tests, genetic testing results, and comprehensive pediatric neuropsychological development assessment. Results:A total of 10 patients were recruited, including 7 males and 3 females, with an age range of 27 days to 5 years and 9 months. 9 patients underwent children′s neuropsychological and behavioral assessments, and the results were consistent with global developmental delay, including 2 mild cases, 4 moderate cases, and 3 severe cases. 3 cases had autism spectrum disorder, and 2 cases had epilepsy. 6 patients underwent complete head MRI examination, and 4 of them showed abnormalities, including delayed myelination, widening of the local extra brain space in the frontal lobe, and abnormal frontal lobe morphology. All 10 cases had point variants. Among them, 9 cases are de novo and 1 case is maternal inheritance. Out of 10 cases, there were 5 cases with copy number variations, but all of them were of unknown significance. Among the 10 variants, 8 have been reported and 2 have not been reported, namely c.4145A>T(p.N1382I) and c.4937T>A(p.I1646N). In this study, 4 out of 10 patients with SCN2A variants had variation sites located in the S4 segment of domain which constitute Nav1.2, the sodium ion channel encoded by SCN2A. The developmental quotient level was lower when the variation sites were located in the S4 segment of domain, and the difference was statistically significant ( t=-3.101, P=0.017), indicating that the severity of developmental delay may be related to the localization of amino acids corresponding to variant sites within the protein domain. Conclusion:SCN2A mutations are strongly associated with diverse neurodevelopmental disorders. In this study, the phenotypic spectrum of SCN2A variants encompassed epilepsy, global developmental delay, and autism spectrum disorder. Affected individuals exhibited early-onset developmental delays, predominantly moderate to severe in severity. Voltage-sensing domain dysfunction in sodium channels may constitute a critical pathomechanism underlying neurodevelopmental impairments. Further electrophysiological characterization and molecular mechanistic studies are warranted todelineate the genotype-phenotype correlations between specific variant loci and clinical severity.

Objective : To explore the characteristics of impaired attention network function in chronic insomnia pa- tients． Methods : Polysomnography ( PSG) ，sleep scale ，attention network test ( ANT) and neuropsychological cognitive scale were used to evaluate the sleep quality，attention network function and cognitive function of 73 pa- tients with chronic insomnia and 65 healthy subjects. Results : Compared with normal control group，the scores of pittsburgh sleep quality index(PSQI) and insomnia severity index (ISI) in the chronic insomnia group increased， total sleep time reduced，sleep latency prolonged，sleep efficiency decreased，wake after sleep onset and arousal index increased，the proportion of non-rapid eye movement sleep stage 1 increased，the proportion of non-rapid eye movement sleep 3 and the proportion of rapid eye movement sleep decreased，and differences were statistically sig- nificant (all P＜0．05) ．In the attention network test，the efficiency of executive control network decreased in the chronic insomnia group，and the difference was statistically significant (all P＜0. 05) ．In neuropsychological tests， patients in the chronic insomnia group had spent more time on the Stroop color word test-word and trail making test- B．Correlation analysis showed that executive control function was associated with decreased non-rapid eye move- ment sleep stage 3 andincreased PSQI scores in chronic insomnia group． Conclusion Chronic insomnia patients have a certain degree of cognitive impairment，mainly executive control network impairment，which is associated with slow-wave sleep reduction.

Objective:To explore the clinical efficacy of the double eyelid plasty with incision in eyelids margin position and internal fixation.Methods:A retrospective analysis was carried out in 47 patients who underwent double eyelid surgery in the outpatient department of our hospital from September 2015 to June 2017. There were 5 males and 42 females, aged from 17 to 32 (25±4) years. The skin was incised above the eyelid margin of 1-2 mm after anesthesia, orbicularis oculi muscle under the eyelid line, loose organization, and orbital septum fat were stripped and removed. The dermis and the tarsus were sutured under the double eyelid line, and the skin was sutured without any tension.Results:After collecting and analyzing 47 cases from September 2015 to June 2017 in our hospital with this method, we found that all 47 patients achievedⅠincision healing, which showed slight swell, natural and smooth radian of double eyelid, and without complications such as infection, hematoma and ptosis. And this method showed no obvious scar hyperplasia around incision line and thus possessed high patient satisfaction.Conclusions:With the advantage of less trauma, quick recovery, unobvious scar, natural and beartiful double eyelids, this method can be used as a supplementary method for the reconstruction of the blepharoplasty, which is worth promoting.

Objective To investigate the clinical features and risk factors of multidrug－resistant bacteria (multi drug resistant organisms, MDROs) infection in Department of Neurosurgery, and to provide evidence for the prevention and control of MDRO infection. Methods Data from 437 cases of infection in hospitalized patients on January 2012－2016 year in December Third Affiliated Hospital of Guizhou Medical University were retrospectively analyzed. Patients were divided into MDROs group and non MDROs group based on the results of MDROs detection. Multi factor Logistic regression analysis model was used to analyze risk factors. Results The infection rate of MDROs was 35.51%, and the detection rate of MDROs was 33.23% . ESBLs, CR－AB and MRSA were the most common bacterial species, and the infection of respiratory tract, urinary tract and wound infection were the main infection sites. Multivariate logistic regression analysis showed that hospitalization time ＞20 d, level of consciousness (coma), occupancy of ICU ≥7 d, ventilation (invasive), number of antibiotics used≥3, combined use of antibiotics≥3, mechanical ventilation Time≥7 d were possible risk factors for MDROs infection in neurosurgical patients (P＜0.05). Conclusion The situation of MDROs infection in neurosurgery is severe. To reduce MDROs infection, it is important to shorten unnecessary hospitalization time, promptly assess and transfer out of ICU as soon as possible, improve microbial examination, avoid frequent change of antibiotics or unnecessary use of use of broad－spectrum antibiotics, reduce unnecessary mechanical ventilation time, change to non－invasive ventilation as far as possible when the condition permits, focus on patients with poor consciousness, and prevent aspiration by mistake.

OBJECTIVE:To provide reference for rational drug use and hospital infection control. METHODS:AmpC enzyme-producing Enterobacter cloacae were isolated from non-sputum specimen of a hospital during Jan. 2011-Oct. 2017. Drug sensitivity test was conducted by using MIC. The situation of AmpC enzyme production was confirmed by three dimensional test, and that of ESBLs-producing stain was detected with double-disk synergy test. RESULTS:There were 546 strains of AmpC enzyme-producing E. cloacae isolated from non-sputum specimen of the hospital,accounting for 4.80% of non-sputum specimen (546/11 375)and 38.97% of E. cloacae(546/1 401). Top 3 non-sputum samples in the list of detection rate were wound secretion (27.29%),midstream urine(25.82%)and blood(21.79%),and the departments with high detection rate were ICU(22.89%), neurosurgery department(18.68%)and general surgery department(16.67%). Resistance rate of AmpC enzyme-producing E. cloacae to most commonly used antibiotics was higher than 40%. There was statistical significance in resistant rate of the bacteria to ceftriaxone, cefotaxime, gentamicin, nitrofurantoin, levofloxacin, piperacillin/tazobactam, cefoperazone, ceftazidime,cefepime,tobramycin and minocycline among different years (P<0.05). The resistant rate to imipenem and meropenem was lower than 2%. Among 546 strains of AmpC enzyme-producing E. cloacae,68 strains of ESBLs were detected,and detection rates were 5.77%,6.06%,8.70%,10.26%,13.79%,17.35%,18.75% during 2011-2017. CONCLUSIONS:AmpC enzyme-producing E. cloacae are mainly isolated from samples as wound secretion and midstream urine,and mainly come from ICU and neurosurgery department. The drug resistance of the bacteria is severe,and drug resistance of the bacteria to antibiotics as β-lactams and quinolones is increased significantly. The detection rate of ESBLs-producing strain increases year by year. The bacteria are sensitive to carbapenems antibiotics,which can be regarded as first choice. It is necessary to strengthen drug resistance and enzyme production monitoring of AmpC enzyme-producing E. cloacae,select antibiotics combined with results of drug sensitivity test so as to prevent or delay the rapid increase of its resistance rate.

Objective To evaluate the antitumor effect of dacarbazine (DTIC) on B16-F1 melanoma after CDK2 gene silencing.Methods Cultured B16-F1 melanoma cells were divided into 4 groups:control group receiving no treatment,CDK2-shRNA group infected with a recombinant lentivirus pUL-CDK2-shRNA,DTIC group cultured in 96-well plates followed 12 hours later by the treatment with 250 μmol/L DTIC,CDK2-shRNA + DTIC group infected with pUL-CDK2-shRNA followed 12 hours later by the treatment with 250 μmol/L DTIC.MTT assay was performed to evaluate the growth inhibition of B16-F1 melanoma cells,and coefficient of drug interaction (CDI) was calculated.AnnexinV-FITC/PI double staining was conducted to detect cell apoptosis.C57BL/6 mice were subcutaneously injected with B16-F1 cells at exponential growth phase into the right groin to establish melanoma-bearing mouse models.Twenty mouse models were randomly and equally divided into 4 groups:control mouse group injected with phosphate-buffered solution (PBS) into tumors,CDK2-shRNA mouse group injected with pUL-CDK2-shRNA into tumors,DTIC mouse group injected with DTIC into the abdominal cavity,and CDK2-shRNA + DTIC mouse group treated with pUL-CDK2-shRNA and DTIC.The animal experiment lasted 18 days,and the tumor growth curve was drawn.After 18-day treatment,all the mice were sacrificed,and tumors were isolated and weighed.The tumor growth inhibition rate was calculated,and the tumor cell apoptosis was detected by terminal deoxynucleotidyl transferase-mediated dUTP nick-end-labeling (TUNEL).Results After 72-hour culture,compared with the control group,the CDK2-shRNA group,DTIC group,and CDK2-shRNA + DTIC group showed significantly decreased relative cell survival rates (40.6％ ± 2.8％,45.2％ ± 3.7％,28.7％ ± 2.1％,respectively;F =458.04,P ＜ 0.05),but significantly increased cell apoptosis rates (25.1％ ± 3.3％,15.6％ ± 2.2％,45.6％ ± 3.5％,respectively;F =115.46,P ＜ 0.05).Additionally,CDK2-shRNA + DTIC group showed significantly lower relative cell survival rates (P ＜ 0.01),but higher cell apoptosis rates (P ＜ 0.01) compared with the DTIC group.The CDI value was less than 0.7.On the sixth day after the in vivo treatment,the tumor volumes in the control mouse group,CDK2-shRNA mouse group,DTIC mouse group,and CDK2-shRNA + DTIC mouse group were (185.44 ± 68.97) mm3,(83.91 ± 14.33) mm3,(123.70 ± 20.85) mm3,and (34.54 ± 10.72) mm3 respectively.From then on,the CDK2-shRNA mouse group,DTIC mouse group,and CDK2-shRNA + DTIC mouse group showed significantly decreased tumor growth rates compared with the control mouse group (F =11.819,P ＜ 0.05),and the tumor growth rate was significantly lower in the CDK2-shRNA + DTIC mouse group than in the DTIC mouse group (P =0.04).The calculated tumor growth inhibition rates in the CDK2-shRNA mouse group,DTIC mouse group and CDK2-shRNA + DTIC mouse group were 52.2％,41.2％ and 86.4％ respectively.Compared with the control mouse group,the CDK2-shRNA mouse group,DTIC mouse group,and CDK2-shRNA + DTIC mouse group showed significantly increased tumor cell apoptosis indice (32.93％ ± 3.72％,21.62％ ± 3.54％,63.29％ ± 4.74％ respectively;F =222.25,P ＜ 0.05).Moreover,the tumor cell apoptosis index was significantly higher in the CDK2-shRNA + DTIC mouse group than in the DTIC mouse group (P ＜ 0.01).Conclusion CDK2 gene silencing can enhance the inhibitory effect of DTIC on the growth of melanoma,and show a synergistic effect with DTIC,likely by increasing the apoptosis of tumor cells.

Objective To study the removal effect of Moringa seedprotein on turbidity water. Methods The protein of Moringaoleifera seed was extracted by salting out and salting out methodand the protein concentration of Moringa oleifera seed wasdetermined by Coomassie blue staining;The removal effect of Moringa seed protein on turbidity water was determined by coagulation test.The contents of COD, ammonia nitrogen and nitrate nitrogen in the water were determined to determine the effect of Moringa seed protein on water quality. Results The experimental results showed that Moringa oleifera seed protein has good removal effect on high and medium turbidity water, and its removal effect is in a dose - dependent manner.The removal rate of 7 mg/L of Moringa seed protein to high and medium turbidity water reached 92.25 ％ and 64.71 ％ respectively. But the removal efficiency of low turbidity water was less than 7 mg/L, the removal rate of low turbidity water was only 31.91％.The results of determination of COD, ammonia nitrogen and nitrate nitrogen showed that the Moringa seed protein did not increase the content of organic matter in the water while removing turbidity effectively. Conclusion Moringa oleifera seed protein has a certain removal effect on turbidity water, among which the removal effect of high and medium turbidity water is strong, and the removal effect of low turbidity water is poor.Moringa oleifera seed protein had little effect on water quality.