Objective To investigate the mechanism of the permeability change in the blood brain barrier(BBB)at early stage of infectious brain edema which was pretreated with LPS.Methods PMNs were isolated and purified.The concentration of Glu was measured in the supernatant without cells.The rat BMECs were primarily isolated and cultured.The permeability of BMECs waB evaluated by measuring the passing rate of 125 I-BSA via γ ray counter.The expression of NMDAR1 on BMECs was evaluated.Results The concentration of Glu in the LPS preconditioning PMNs subgroup at 5 rain time point was highest.In each group conditioned for 240 min,the permeability index of Bake average of the LPS preconditioning PMNs extract group was highest(P＜0.01).The expression of NMDARI increased in the LPS preconditioning PMNs extract group was more than that in other groups.Conclusions The research showed for the first time that the concentration of Glu increased after LPS preconditioning and the Glu excreted by PMNs can promote the permeability of BMECs and change the function of BBB,which possibly may be related the mechanism of NMDAR1 expression increased in BMECs.

OBJECTIVETo explore the clinical effects of lattice ultra pulse carbon dioxide laser combined with traditional Chinese medicine ( Fuchunsan ) on the treatment of postburn hyperplastic scar.

METHODSSixty-three patients with hyperplastic scar after burn injury hospitalized from February 2012 to June 2014 in our department were treated with lattice ultra pulse carbon dioxide laser combined with traditional Chinese medicine (Fuchunsan). Patients were divided into early stage group (E, n = 35), middle stage group (M, n = 25), and late stage group ( L, n = 3) according to the formation time of scar, which was respectively 3 weeks to 3 months, longer than 3 months and less than or equal to 6 months, and 3 to 15 years in groups E, M, and L. The number of times of laser treatment of patients in each group was recorded. The degree of scar pain in patients of the three groups was assessed by the Numerical Rating Scale (NRS) before treatment and after treatment for 1, 2, and 3 times. The scar condition of patients in groups E and M was assessed by the Vancouver Scar Scale (VSS) before treatment and after treatment for 1, 3, and 5 times. Patients in group L did not receive VSS assessment but were evaluated by clinical observation only. Photos of scar in treating area were taken before treatment and after treatment for 3 and 5 times to evaluate the clinical effect. Data were processed with t test.

RESULTSPatients in groups E and M were treated with laser for (4.8 ± 1.1) and (7.7 ± 2.1) times respectively. In group L, the treatment was stopped in 2 patients after laser treatment for 5 times, and 1 patient received laser treatment for 12 times. The degree of pain in patients of groups E and M was alleviated significantly after treatment for one time, and the number of patients scoring 1-4 point(s) in NRS increased from 5 cases to 38 cases. After treatment for 2 and 3 times, the increase in the number of patients scoring 1-4 point (s) in NRS was on a small scale. Before treatment and after treatment for 1 time, VSS scores of patients in groups E and M were similar (with values respectively 0.641 and 0. 082, P values above 0. 05). After treatment for 3 and 5 times, VSS scores of patients in group E were respectively (9.2 ± 0.8) and (7.0 ± 1.1) points, which were significantly lower than those in group M [ (9.7 ± 1.0) and (8.2 ± 1.0) points, with values respectively -1.993 and -4.433 , P < 0.05 or P < 0.01]. After treatment for 3 times, the rate of improvement in appearance was respectively 88.6% (31/35) and 72.0% (18/25) in groups E and M, and it was respectively 100.0% (35/35) and 96.0% (24/25) after treatment for 5 times. No significant effect in appearance was found in the 3 patients in group L.

CONCLUSIONSEarly application of lattice ultra pulse carbon dioxide laser combined with traditional Chinese medicine (Fuchunsan) for the treatment of postburn hyperplastic scar is effective.

Burns ; Cicatrix ; etiology ; prevention & control ; Humans ; Lasers, Gas ; therapeutic use ; Medicine, Chinese Traditional ; Postoperative Care ; methods ; Postoperative Complications ; prevention & control ; Treatment Outcome

Aim Toclarifytheeffectofacteosideon proliferation of neural stem cells (NSCs ) from adult mice,as well as the involved signaling pathway.Meth-ods NSCswereisolatedfromthesubventricularzone (SVZ)of adult C57BL/6 mice,then identified by im-munofluorescence staining with Nestin,the marker of NSCs.NSCs were exposed to acteoside (5,10,20,40μmol·L-1 )in absence of mitogen(EGF/bFGF)for 24 h.We employed CCK8 assay to detect NSCs viability and BrdU staining to identify NSCs proliferation.We performed Western blot to quantify the expression level ofp-AktinducedbyacteosideonNSCs.Results With-out mitogen,acteoside increased NSCs proliferation by activating p-Akt,which can be blocked by LY294002, the inhibitor of PI3K/AKT signaling pathway.Conclu-sion ActeosidepromotestheproliferationofNSCsfrom adult mice by activating PI3K/AKT pathway.

Objective To explore the correlation between histological chorioamnionitis (HC) and brain injury in preterm infants. Methods Three hundred and forty-seven cases of infants at the gestational age of 28-31 weeks who were admitted to the neonatology department of our hospital were analyzed retrospectively. They were divided into the HC group and the control group according to the pathological examination. Moreover, HC group was divided into FV group and non-FV group according to the pathological findings of fetal vasculitis (FV). Based on the findings of periodical ultrasonography, the incidences of periventricular leukomalacia (PVL), periventricular-intraventricular hemorrhage (PVH-IVH), and the PVL+PVH-IVH were compared among groups. Results The incidences of PVL in the HC group and the control group were 17.9% and 10.3%respectively. The incidences of PVL+PVH-IVH in the two groups were 5.5%and 1.48%respectively, and the difference between two groups was signiifcant (P<0.05). The incidences of PVH-IVH in the two groups were 28.9%and 26.2%respectively, and the difference between two groups was not signiifcant (P>0.05). In the HC group, the incidences of PVL in FV group and non-FV group were 28.1%and 9.87%respectively, and the difference between two groups was signiifcant (P<0.05). The incidences of PVH-IVH in FV group and non-FV group were 34.3%and 24.7%respectively, and the difference between two groups was not signiifcant (P>0.05). The incidences of PVL+PVH-IVH in FV group and non-FV group were 7.81%and 3.70%respectively, and the difference between the two groups was not have signiifcant (P>0.05). Conclusions HC may increase the ncidences of PVL and PVL+PVH-IVH in the preterm infants, while its effect is minimal on PVH-IVH. FV could increase the incidence of brain injury in preterm infants.

Objective To compare two different methods to detect the differences of gene mutation rate, sensitivity, specificity and coincidence rate of epidermal growth factor receptor (EGFR) in non-small-cell lung cancer (NSCLC) so as to assess the clinical value of qRT-PCR method and its environmental-friendly technologyplatforms.One uses environmental fixative poly hydroxyl acrylic acid and green transparent liquid dewaxing Van-clear alone or in combination to replace the traditional fixative 4％ (volume fraction) neutral buffered formalin and the traditional transparent dewaxing liquid xylene in application of quantitative real-time polymerase chain reaction (qRT-PCR).The other uses traditional reagents in direct sequencing.Methods We selected 91 cases of primary NSCLC specimens resected between May 2013 and March 2016 in Zhongshan Bo`ai Hospital and Zhongshan Hospital of Traditional Chinese Medicine.Five samples were taken from the same tumor lesion.We used a random number table to randomly divide these samples into Groups A, B , C, D, and E.Group A received direct sequencing method in detection of EGFR gene mutations.Besides, during the experiment, 4％ neutral buffered formalin was used for fixing, and xylene transparent dewaxing was used to make slices for DNA extraction dewaxing.Group B received qRT-PCR method to detect EGFR gene mutations.Meanwhile, during the experiment, 4％ neutral buffered formalin was used for fixing, and xylene transparent dewaxing was used to make slices for DNA extraction dewaxing.Group C received qRT-PCR method in detection of EGFR gene mutations.At the same time, during the experiment, polyhydroxy acrylic acid was used for fixing, and xylene transparent dewaxing was used to make slices for DNA extraction dewaxing.Group D received qRT-PCR method to detect EGFR gene mutations.In the meantime, 4％ neutral buffered formalin was used for fixing, Van-clear transparent dewaxing was used to make slices for DNA extraction dewaxing.Group E received qRT-PCR method in detection of EGFR gene mutations.In addition, during the experiment, polyhydroxy acrylic acid was used for fixing, and Van-clear transparent dewaxing was used to make slices for DNA extraction dewaxing.In addition, during the experiment, polyhydroxy acrylic acid was used for fixing, and Van-clear transparent dewaxing was used to make slices for DNA extraction dewaxing.The mutations of Exons 18, 19, 20, and 21 in EGFR genes were respectively determined in the five groups of NSCLC.Results ① Groups B, C, D, E and A did not significantly differ in the percentage of people with mutations or target site mutation rates of EGFR genes in NSCLC (P> 0.05).② The detection results of EGFR target site mutation in Groups B, C, D, E and A had good sensitivity, strong specificity, and high compliance rate.Conclusion The green transparent liquid dewaxing Van-clear alone or in combination to replace the traditional fixative 4％ neutral buffered formalin and the traditional transparent dewaxing liquid xylene in the application of qRT-PCR so as to detect EGFR gene mutations in NSCLC has good consistent results compared with the method that uses traditional reagents in direct sequencing.It has the significance and value in clinical application.

Objective To determine the genotype and clustering characteristics of multidrug-resistant (MDR) mycobacterium tuberculosis in Wuhan City. Methods A total of 149 mycobacterium tuberculosis strains from MDR tuberculosis (MDR-TB) patients were genotyped by DTM-PCR, mycobacterial interspersed repetitive unit–variable number tandem repeat typing (MIRU-VNTR) and sequencing of drug-resistant associated genes. The Hunter-Gaston discriminatory index (HGDI), clustering rate and recent minimum estimated infection rate were calculated. Results The cumulative HGDI of all MIRU loci was 0.9944. Gene locus with the highest HGDI（0.6467）was QUB11b. Combing MIRU-VNTR and DTM-PCR, 111 unique genotypes and 11 clustered genotypes（38 strains）from 149 isolates were identified. The clustering rate was 25.50%, and the recent minimum infection rate was estimated to be 18.12%. After further genotyping by sequencing of drug-resistant associated genes, the results showed that 17 (44.74%) strains in each cluster shared the same mutation sites, while the other strains had different mutation sites. Conclusion The combination of DTM-PCR, 24 locus MIRU-VNTR typing and drug resistance gene sequencing demonstrates a relatively high discriminatory power, which is suitable for the genotyping of multi drug resistant mycobacterium tuberculosis in Wuhan. The epidemic of MDR-TB in Wuhan is mainly caused by reactivation, and the recent transmission rate is not high.

Occupational exposure limits for hazardous agents in the workplace—Part 1: Chemical hazardous agents (GBZ 2.1—2019) has been implemented since April 1, 2020. The document redefines the evaluation of occupational exposure by introducing a "reduction factor" to adjust the occupational exposure limits of chemical hazardous agents for long working hour systems to ensure a protection level reasonable for the workers and equivalent to conventional occupational exposure limits. This paper discussed common problems encountered in the context of using this adjustment strategy, such as the adjustment being unable to cover all working hours, and the adjustment of values of terms associated with occupational exposure limits. Corresponding solutions were proposed. Working cycle and average exposure time of each working cycle were introduced to evaluate the actual working hours of workers. Regarding involved terms, adjustments of their corresponding values were clarified according to their definitions and application scenarios. Examples were provided for occupational health service providers so that their occupational health practices could be carried out in a more scientific and reasonable manner.

OBJECTIVETo investigate the effects of the volume of 4% neutral phosphate buffered formalin fixative solution on the detection of human epidermal growth factor receptor 2 () gene by fluorescencehybridization (FISH) in primary invasive breast cancer.

METHODSTissue samples were collected from 109 patients with primary invasive breast cancer admitted in Zhongshan Boai Hospital from June 2014 to October 2016. The ratios of 4% phosphate buffered formalin fixative solution to sample volume samples were 3:1, 6:1, 9:1, 10:1, 15:1, 20:1 or 25:1 (groups A, B, C, D, E, F and G), respectively. Paraffin sections were made after 15 h of fixation. The amplification ofgene was detected by FISH. The gene amplification results ofwere observed and compared in different groups.

RESULTSFluorescence microscope showed that the tissue contour in groups A, B and C was vague, cell debris appeared, and the probe was positioned poorly; while the tissue contour was clear and complete in groups D, E, F and G and the probe was positioned accurately. The positive rate ofwas gradually increased from group A to D(=8.601,<0.01), and that remained stable at 24.77% in groups D to G. The positive rate of gene amplification in groups D, E, F and G was significantly higher than that in groups A, B and C (all<0.05).

CONCLUSIONSWhen using FISH to detectgene in samples of primary breast invasive carcinoma, the volume of fixative solution should be at least 10 times of the sample volume to obtain accurate and stable results.

OBJECTIVE: To investigate paraffin section thickness in immunohistochemical detection of p16 expression in cervical tissue samples. METHODS: p16 immunohistochemical staining was performed in 150 cases of chronic cervicitis, 126 cases of low-grade squamous intraepithelial lesions(LSIL), 96 cases of high-grade squamous intraepithelial lesions (HSIL) and 78 cases of cervical cancer from January 2014 to March 2018 in Zhongshan Boai Hospital. The results of p16 protein expression in paraffin sections with thickness of 2, 3, 4, 5 and 6 μm were compared using Logistic regression analysis. RESULTS: With the increase of slice thickness, the staining of p16 protein in nucleus gradually increased. The thickness of cervical slices in chronic cervicitis and cervical cancer samples had no significant effect on the positive rate of p16 protein(=7.817 and 1.332, both >0.05), while the thickness of slices in LSIL and HSIL samples had significant effect on the positive rate of p16 protein (=17.688 and 10.182, <0.05 or <0.01). The stable and reliable results were obtained when the slices were between 3 and 5 μm thick. CONCLUSIONS Paraffin sections with thickness of 3.0-5.0 μm are recommended for immnohistochemical staining of p16 protein in cervical tissue samples.
Biomarkers, Tumor ; genetics ; metabolism ; Cervical Intraepithelial Neoplasia ; Cyclin-Dependent Kinase Inhibitor p16 ; genetics ; metabolism ; Female ; Histocytological Preparation Techniques ; standards ; Humans ; Immunohistochemistry ; Paraffin ; Squamous Intraepithelial Lesions of the Cervix ; Uterine Cervical Neoplasms ; physiopathology