Objective To study the effect of conditioned media for rat bone marrow mesenchymal stem cells (BMSCs-CM) on palmitic acid (PA)-induced insulin resistance (IR) in HepG2 cells and its underlying molecular mechanisms.Methods HepG2 cells were treated with or without BMSCs-CM and L-DMEM in the presence or absence of PA.Glucose utilization in HepG2 cells were detected with PAS,glucose and glycogen measurements.Western blotting was used to assess the expression of phospho-insulin receptor substrate (p-IRS),phosphatidylinositol 3-kinase (PI3 K) and p-AKT.Results (1) Incubation of HepG2 cells with 0.25 mmol/L PA for 24 hours significantly increased the glucose concentration and decreased the glycogen content (P ＜ 0.05) in the media.(2) Treatment with BMSCs-CM significantly ameliorated the glucose and glycogen alteration in cells pretreated with PA (P ＜ 0.05),however,no obvious effect of BMSCs-CM on the cell glucose and glycogen production.(3) BMSCs-CM treatment also increased protein expression of p-IRS,PI3K and p-AKT in PA incubated HapG2 cells (P＜ 0.05).The effect of BMSCs-CM on PI3K and p-AKT expression could be mimicked upon addition of 740Y-P,a PI3K agonist,but abolished by LY294002,a PI3K specific inhibitor.Conclusions BMSCs-CM could improve the insulin sensitivity in HepG2 cells pretreated with PA through upregulation of insulin signaling component expression.

Objective To investigate the feasibility of congenital cytomegalovirus (CMV)infection screening by saliva polymerase chain reaction.Methods From November 1,2010 to February 29,2012,6733 newborns born in Changzhou Maternal and Child Health Care Hospital were enrolled.Saliva samples (0.2 ml) were collected within 3 days after birth,CMV-DNA was detected by real time-polymerase chain reaction and hearing screening was done with EroScan transient-evoked otoacoustic emissions at the same time.The positive rate of congenital CMV infection screening was calculated and clinical manifestations were analyzed.Chi square test was applied to statistical analysis.Results Totally 6733 newborns were screened and 107 of them were found to be positive with CMV DNA,the positive rate was 1.59％ (107/6733),among which 88 were asymptomatic (82.2％) and 19 were symptomatic (17.8 ％).The major clinical manifestations of the neonates with positive CMVDNA were pathological jaundice (13 cases),hepatomegaly (5 cases),granulocytopenia,thrombocytopenic purpura,anemia and small for gestational age (two cases each).Fourteen newborns had only one major clinical manifestation,three newborns had two major clinical manifestations and two newborns had three major clinical manifestations.There was no statistical difference between newborns with positive and negative CMV DNA on hearing screening [hearing loss in one ear:8.4％ (9/107) vs 5.8％ (382/6626); hearing loss in two ears:3.7 ％ (4/107) vs 2.4 ％ (159/6626),x2 =2.776,P=0.241].Conclusion It is feasible to screen congenital CMV infection with saliva sample.

ObjectiveTo evaluate prognosis and its clinical factors in patients with primary pontine hemorrhage. Methods Patients with primary pontine hemorrhage who were hospitalized in the First Affiliated Hospital of Wenzhou Medical College within 24 hours after stroke onset between April 2007 and April 2009 were registered conscutively. The patients were followed up for one year. Kaplan-Meier methods were used to analyze survival rate. Cox proportional hazards model was used to study risk factors for 1-year mortality. ResultsA total of 41 patients with primary pontine hemorrhage were studied. Their mean age was (63.5 ± 10. 1 ) years.The overall 1-year mortality rate was 61.0％, the median survival time was (80. 0 ±54.4) days (95％ CI 0-186. 64). After one-year follow-up, the mortality rate in patients with primary dorsal pontine hemorrhage( 18.2％ ) was significantly lower than that in patients with primary ventral pontine hemorrhage(72. 7％ ; x2 = 8. 800, P = 0. 003 ). Patients with massive primary pontine hemorrhage had significantly higher mortality rate than patients with dorsal primary pontine hemorrhage( x2 = 8. 927, P =0. 003). The average hematoma volume of the survivor group and mortality group was (3. 043 ± 1. 718) ml and (5. 984 ± 2. 707) ml, respectively, showing statistical significance (t = 3. 661, P = 0. 001 ). Analysis with Cox proportional hazards model showed that the risk factors associated with mortality were hematoma location ( RR = 2. 428, 95 ％ CI 1. 055-5. 587 ), hematoma volume ( RR = 1. 283, 95 ％ CI 1. 044-1. 577 ),GCS score on admission(RR =3. 389, 95％ CI 1. 177-9. 756). Patients with pontine hematomas in dorsal had a significantly better outcome than in other locations.Conclusions The survival and prognosis in primary dorsal pontine hemorrhage are better than with hemorrhaging in other parts of pontine. A significant correlation was observed between poor prognosis and hematoma volume, hematoma location and GCS score on admission.

Carbon nanotubes/Au nanoparticles ( CNT/AuNP ) composite film was fabricated on glassy carbon electrode ( GCE) by first dropping CNTs on the electrode surface and then electrodeposition of AuNPs by multi-potential step. The antibody of microcystin-( leucine-arginine ) ( anti-MCLR ) was immobilized on the modified electrode surface through adsorption on AuNPs. Subsequently, bovine serum albumin ( BSA) was used to block the non-specific adsorption to obtain the immunosensor for MCLR assay. The immunosensor could effectively capture MCLR by the specific immunoreaction between the electrode surface-confined antibody and MCLR, followed by the attachment of the anti-MCLR HRP-labeled to form a sandwich-type system. The analysis of MCLR was performed based on the catalytic reaction of HRP toward the oxidation of hydroquinone ( QH2 ) by H2 O2 . Under the optimal experimental conditions, the peak current response increased linearly with the concentration of MCLR in the range of 0 . 50-12 μg/L with a detection limit of 0. 30 μg/L (S/N=3). The developed immunosensor was used to determine MCLR in real water samples, and the recoveries of standard addition experiments were in the range of 93 . 0%-108 . 5%, with the relative standard deviation of 3 . 8%-5 . 0%.

Objective:To explore the method of establishing a modified demyelination and myelination regeneration model induced by dicyclohexanone oxalyl dihydrazone (CPZ) in mice with multiple sclerosis (MS), and to analyze the image markers of demyelination and myelination regeneration in mouse MS model.Methods:After the intragastrically administered with sodium carboxymethyl cellulose (CMCNa) for one week, a total of 30 C57BL/6 male mice were randomly divided into the control group ( n=10), the demyelination group ( n=10), and the remyelination group ( n=10). The mice of the control group were immediately performed MR scanning and pathological specimen obtaining; the mice in the demyelination group were administered with intragastrical CPZ-CMCNa once a day for 6 weeks for inducing demyelination, then received MR scanning and specimen obtaining with the same protocols used in control group; the mice in the remyelination group were administered with intragastrical CPZ-CMCNa once a day for six weeks for demyelination, then CPZ was withdrawn and normal diet was given for another four weeks. Then MR scanning and specimen obtaining were performed with the same protocols used in the other two groups. Regions of interest (ROIs) were set at the rostrum of corpus callosum (rCC), the bilateral normal appearing white matters (NAWM) of the rostrum of corpus callosum, and the bilateral cerebral cortex (Cx). The normalized T 2WI (T 2-normalized), fractional anisotropy (FA), mean diffusivity (MD), axial diffusivity (AD), and radial diffusivity (RD) values were compared among the three groups by one-way ANOVA. Results:The demyelination and remyelination mice model of MS were successfully established. The T 2-normalized values of rCC in control group, demyelination group and remyelination group were 0.47±0.03, 0.72±0.04, 0.54±0.04, respectively, with statistically significant difference found ( F=90.511, P<0.05). Post-hoc multiple comparisons showed significant differences among those groups ( P<0.05). There was no significant difference of T 2-normalized value in NAWM and Cx among the three groups ( P>0.05). Moreover, there were significant differences in the FA values (0.36±0.04, 0.29±0.03, and 0.32±0.05), the MD values [(0.572±0.015), (0.598±0.034), and (0.626±0.043)×10 -3 mm 2/s], the AD values [(0.79±0.04), (0.77±0.06), and (0.83±0.04)×10 -3 mm 2/s], and the RD values [(0.46±0.02), (0.51±0.03), and (0.53±0.05)×10 -3 mm 2/s] of rCC of the control group, the demyelination group, and the remyelination group (all P<0.05). Significant difference was found in FA values between the demyelination group and the control group ( P<0.05), and in MD values between the remyelination group and the control group ( P<0.05), as well as in AD values between the remyelination group and the demyelination group ( P<0.05). There were also significant differences in RD values between the remyelination group and the control group, and the demyelination group and the control group (all P<0.05). However, no significant difference was found in all diffusion tensor imaging (DTI) metrics of NAWM and Cx among the three groups (all P>0.05). The LFB-eosin staining showed that the myelin sheath of rCC was lost in the demyelination group, and the rCC was partially regenerated and repaired in the remyelination group. Conclusion:The modified CPZ-CMCNa model can selectively induce demyelination and remyelination of rCC, and the changes of demyelination and remyelination of rCC in the modified CPZ-CMCNa model can be quantitatively detected by T 2WI combined with DTI, which might provide related theoretical basis for the study on dynamic changes of MS lesions.

Objective:To explore the value of Neoseq in screening and diagnosis of neonatal fatty acid oxidation disorders (FAOD).Methods:A retrospective case-control study was conducted on 163 500 live births in Changzhou city from April 2015 to April 2021. The following two models were adopted for FAOD screening and diagnosis. (1) Traditional mode: Heel blood samples were obtained from all subjects for initial screening using tandem mass spectrum (TMS), followed by next-generation sequencing (NGS) and other differential diagnostic testings for those with positive results. (2) Neoseq: Neoseq was performed on the true positive, negative and false positive cases according to the traditional mode screening results. The detection rate, additional discovery, reporting period, and other parameters of the two models for FAOD were described and compared.Results:(1) Detection and diagnosis of FAOD: A total of 18 confirmed cases of FAOD were detected through the traditional model, with an incidence of 1/9 083 in Changzhou city. The positive rate was 0.55% (907/163 500) for initial TMS and 0.04% (73/163 500) for the second. The positive predictive value was 2.0%(18/907), with a false positive rate of 98%(889/907) in the initial screening. (2) The results of Neoseq: ①Pathogenic mutations were detected in 16 of the 18 confirmed cases, and the coincidence rate of mutation sites between the two methods was 16/18. The other two confirmed cases were missed diagnosed by Neoseq, including one β-ketothiolase deficiency with only one detected pathogenic mutation and one medium-chain acyl-CoA dehydrogenase deficiency without any detected pathogenic mutation. ②No pathogenic mutations were detected in the 57 false-positive cases by Neoseq. ③Among the 100 negative cases in initial screening, DUOX2 heterozygous mutation, and MTTL1 hemizygous mutation were detected in one case each. ④The median period of results reporting was 43.5 d (28-104 d) for the traditional mode and 12 d (10-15 d) for the Neoseq mode. Conclusions:Neoseq has a high detection rate for FAOD. Combined with TMS screening, Neoseq reduces the false-positive rate of biochemical screening, rapidly identifies genetic causes by shortening the results waiting time and covers diseases that couldn't be detected by traditional biochemical methods.

Objective To investigate the influences of maternal gestational diabetes mellitus (GDM) on amino acid levels in neonates.Methods From June 2016 to May 2017,393 pregnancies diagnosed with GDM in Changzhou Maternity and Child Health Care Hospital Affiliated to Nanjing Medical University and 3 924 normal pregnancies were enrolled in this study.Clinical data of the gravidas and their newborns were collected.Heel blood samples were collected at 72 to 96 hours after birth.Tandem mass spectrometry was performed to detect the levels of 11 amino acids including alanine,arginine,citrulline,glycine,leucine/ isoleucine/hydroxyproline,methionine,ornithine,phenylalanine,proline,tyrosine and valine in neonatal heel blood.Differences in amino acid levels between the two groups were compared by t test.Influences of GDM on neonatal amino acid levels were analyzed by multivariate linear regression.Results Compared with the healthy pregnancy group,neonates in the GDM group had higher levels of methionine [(21.01 ±6.30) vs (19.93±6.47) μmol/L,t=3.159,P=0.002] and phenylalanine [(47.19±9.19) vs (45.78±8.58) μ mol/L,t=3.076,P=0.002],but lower levels of alanine [(280.51 ±64.54) vs (290.15±68.40) μ mol/L,t=2.678,P=0.007],proline [(147.64±30.64) vs (152.36±33.57) μ mol/L,t=2.680,P=0.007],tyrosine [(85.21 ±29.50) vs (90.60± 33.32) μ mol/L,t=3.089,P=0.002] and ornithine [(101.22±28.79) vs (105.83±30.10) μmol/L,t=2.906,P=0.004].Multivariate linear regression analysis showed that GDM was responsible for the increase of methionine (β=0.69,95％CI:0.02 to 1.37,P=0.044) and phenylalanine (β=1.60,95％CI:0.69 to 2.51,P=0.001),and the decrease of tyrosine (β=-4.98,95％CI:-8.42 to-1.54,P=0.005) and ornithine (β=-3.16,95％CI:-6.30 to-0.02,P=0.048) in neonates.Conclusions GDM neonates has increased of methionine and phenylalanine levels and decreased tyrosine and omithine levels.

Endometriosis（EMs） is a common chronic inflammatory gynecological disease，affecting about 5%-10% of women of childbearing age worldwide，and has always been a major challenge in clinical treatment. Huposan， derived from the Experiential Prescriptions for Universal Relief （《普济本事方》）， has the effects of moving qi， activating blood，expelling blood stasis， and relieving pain. It is often used to treat EMs clinically and has achieved good curative effect. The relevant studies on the treatment of EMs by Huposan were retrieved from databases， such as CNKI，PubMed，Wanfang Data， and VIP for summarizing the mechanisms of action and clinical application of Huposan in the treatment of EMs， aiming to provide ideas and references for the basic research and clinical application of Huposan. As revealed by basic experiments，Huposan could exert therapeutic effects on EMs through resisting cell adhesion by reducing intercellular adhesion molecule 1（ICAM-1）content，decreasing concentrations of matrix metalloproteinase（MMP）-2 and MMP-9 against ectopic endometrial invasion，inhibiting vascular endothelial growth factor（VEGF）expression for antiangiogenesis，inhibiting the expression of tumor necrosis factor-α（TNF-α）， interleukin （IL）-6， and IL-1，reversing helper T cell（Th）1/Th2 balance shifts to regulate the body's immune mechanism，and reducing the serum levels of nitric oxide（NO）and nitric oxide synthase（NOS）. In clinic practice，Huposan has the effects of reducing ectopic lesions，relieving pain symptoms，reducing serum carbohydrate antigen 125（CA125）content，improving hormone levels，regulating endocrine and immune factors，and reducing postoperative recurrence rate. Huposan plays a therapeutic role in EMs through multiple targets and mechanisms，which is worthy of further exploration and clinical promotion.

Autophagy is a lysosome-mediated catabolic process that captures and degrades dysfunctional organelles and useless proteins during cellular stress process， which plays a dual role in cervical cancer. In the early stage of cervical cancer， autophagy inhibits the occurrence and development of cervical cancer by prohibiting the accumulation of oncogenic p62 protein. In the advanced stage of cervical cancer， inhibition of autophagy of cancer cells enhances the sensitivity of cancer cells to chemotherapeutic drugs， thus inhibiting their proliferation. In recent years， the research on Chinese medicine monomers regulating autophagy in the treatment of cervical cancer has attracted extensive attention from scholars at home and abroad. Chinese medicine monomers regulate the autophagy of cervical cancer cells through multiple pathways and multiple targets， so as to increase the apoptosis rate and reduce the resistance of cancer cells to chemotherapeutic drugs. Therefore， this paper reviewed the mechanism of Chinese medicine monomers in inhibiting cervical cancer through autophagy， expecting to find new breakthroughs in the discovery and development of preventive and therapeutic drugs for cervical cancer. By reviewing the literature， it was found that in the early stage of cervical cancer， Chinese medicine monomers activated autophagy to promote apoptosis of cancer cells， and the main mechanism was to increase lysosomal membrane permeability and chemotherapeutic sensitivity and activate intact autophagy flow. In the advanced stage of cervical cancer， inhibition of autophagy reduced the sensitivity of cancer cells to chemotherapy drugs by inhibiting the formation of autophagosomes and autolysosomes. The treatment of cervical cancer by Chinese medicine monomers regulating autophagy has achieved certain effect， but there are few clinical experimental studies and lack of reliable clinical theoretical basis. Therefore， it is essential to carry out more clinical experimental studies on Chinese medicine monomers regulating autophagy to treat cervical cancer， thus finding more reliable theoretical basis for the treatment of tumors.

Objective:To provide epidemiological data and clinical evidence for cosmetic adverse reactions.Methods:A retrospective clinical analysis was carried out on a total 820 outpatients (23 males and 797 females) suspected to be with cosmetic adverse reactions from January 2014 - October 2017, and average age of these patients was 7~75 (32.66±8.09) years. Suspicious cosmetics patch tests were performed in some patients. Suspicious cosmetics patch tests were performed in 687 patients.Results:Among 820 patients with cosmetic adverse reactions, women accounted for 97.20% and men accounted for 2.80%. Age distribution was most common among young people aged 21-40 years, accounting for 71.34%. The highest level of education was higher education, accounting for 59.69%. Occupational distribution was most commonly concentrated in employees and unemployed persons, accounting for 28.54% and 18.66%, respectively. A history of cosmetics allergies accounted for 17.28%. Cosmetic contact dermatitis was the most common clinical type of cosmetic adverse reactions, accounting for 92.70%. A total of 1682 suspected pathogenic cosmetics were involved. The positive rate of the cosmetic original patch test was 42.39%. Among the cosmetics with a positive patch test, moisturizing, anti-wrinkle and whitening freckle cosmetics accounted for the highest proportion, 31.59%, 15.09%, and 12.68%, respectively.Conclusions:Cosmetic contact dermatitis is the most common type of cosmetic adverse reaction. Patch testing is helpful in identifying the contact allergens in cosmetic adverse reaction.