OBJECTIVETo investigate the functions of human periodontal myofibroblast (MFB) in vitro.

METHODSHuman periodontal fibroblast (hPDLFs) was cultured and induced to MFB by transforming growth factor-β1 (TGF-β1). MFB was denoted as the experimental group, whereas the hPDLFs was the control group. The groups were continuously cultured and harvested at 0, 12, 24, 48, and 72 h. The MFB marker α-smooth muscle actin (α-SMA) was examined by immunocytochemistry. The expression of fibronectin (FN) between MFB was examined by immunocytochemistry to detect the MFB contact relationship. The mRNA expression levels of α-SMA, collagen (Col) I, and Col III were measured by reverse transcription-polymerase chain reaction (RT-PCT) to analyze extracellular matrix secretion. The protein expression levels of α-SMA and Col I were also assessed by Western blot.

RESULTSThe experimental group had significantly higher α-SMA expression than the control group at 0 h (P < 0.001). A positive expression of FN was found between MFB. The experimental group had significantly higher expression levels of Col I and Col III than the control group at 24 h (P < 0.001).

CONCLUSIONHuman periodontal MFB presents a continuous, high expression of α-SMA. MFB could interact through FN. MFB is significantly capable of extracellular matrix secretion.

Actins ; Epithelial Cells ; Extracellular Matrix ; Fibroblasts ; Fibronectins ; Humans ; Jaw ; metabolism ; Myofibroblasts ; Transforming Growth Factor beta1

Object To study the expression of miRNA-146a in sepsis-induced acute lung injury (ALI) patients and its effect on the inflammation in mouse models.Methods miRNA-146a expression in peripheral blood was determined in sepsisinduced ALI patients and healthy volunteers by qRT-PCR.Sepsis-induced ALI mouse model were reproduced by LPS treatment and miRNA-146a mimic,miRNA-146a NC and miRNA-146a inhibitors were injected through trachea.The expressions of miRNA-146a,TNF-α,IL-1β and COX-2 mRNA were determined by qRT-PCR 24h after the intervention.Results The miRNA-146a expression in peripheral blood significantly increased in severe sepsis with ALI patients,compared with the control subjects.For mouse model,the expressions of TNF-o,IL-1β and COX-2 mRNA significantly decreased in miRNA-146a group,while increased in miRNA-146a inhibitor group compared with miRNA-146a NC group (P＜0.05).The TNF-α and IL-1β expressions significantly decreased in miRNA-146a inhibitor+COX-2 inhibitor group compared with miRNA-146a inhibitor group (P＜0.05).Conclusion MiR-146a treatment can effectively alleviate the lung inflammation in sepsis-induced ALI mice.

Objective:To clone and express panallergen profilin from the pollen of coco(Cocos nucifera Linnaeus).Methods:RT-PCR and RACE methods were applied to clone the full-length panallergen genes from coco pollen and the sequence was analyzed.The specific primers were designed.The ORF of profilin of coco pollen was amplified with RT-PCR and cloned into the expression vector pET 28a.Expression of the recombinant coco pollen profilin was carried out in E.coli BL21(DE3) and the purification of the recombinant protein was performed via affinity chromatography with Ni2+ coupled to sepharose.IgE reactivity to recombinant coco pollen profilin was investigated by immunoblot.Results:The complete sequence of coco pollen profilin was cloned.The sequence was 608 bp and included an open reading frame(396 bp) coding for 131 amino acids.Sequence analysis showed that the deduced protein was an acidic protein with an estimated molecular mass of 14.19 kD and a pI of 4.61.The GeneBank accession number of the clones was EF173598.After overexpressed in E.coli BL21(DE3),the recombinant protein was purified through affinity chromatography with Ni2+ coupled to sepharose.Immunoassay showed that the recombinant allergen has good IgE binding capacity.Conclusion:The profilin of coco pollen is expressed successfully in BL21(DE3),which will be used as a base for further study on coco pollen related allergy.

AIM: To know the variations of the cytochrome b gene in cancer tissue, paracarinoma tissue and normal tissue and to inquire into the relationship between mutations of mitochondrial genome and carcinogenesis. METHODS: Cellular total DNA was extracted.The cytochrome b genes of three tissues were amplifyed with polymerase chain reaction(PCR). PCR products were analysed by DNA auto-sequencing method. RESULTS: The cytochrome b gene of cancer tissue had the C to G mutation at nt 14931, the C to G mutation at nt 15004 and the T to C mutation at nt15435,respectively. The cytochrome b gene of paracarinoma tissue had the A to C mutation at nt 15436. The cytochrome b gene of normal tissue had not mutation. CONCLUSION: Mitochondrial DNA mutations could be the endogenous factors that induce nuclear genome mutation. It could promoto carcinogenesis. The paracarinoma tissue was abnormal in DNA molecular level.

Abstract: Objective To compare the application value of metagenomic next generation sequencing (mNGS) with traditional culture in diagnosis of pulmonary infection pathogens. Methods　The clinical documents of 310 patients with suspected pulmonary infection admitted to the General Hospital of Center Theater Command from February 2021 to September 2022 were retrospectively analyzed. The results of mNGS and traditional culture were analyzed, followed by comparison on the positive rate, sensitivity, specificity, accuracy (ACC), positive predictive value (PPV) and negative predictive value (NPV) between the two methods. Results　 The study revealed that mNGS can simultaneously detect multiple pathogens, with the highest efficiency of detection for bacteria and the lowest for fungi. And the sequencing numbers of bacteria, fungi and viruses shown by mNGS were significantly different (H=70.361, P<0.001). In comparison, mNGS displayed a higher positive detection rate (88.40%) than traditional culture (29.70%) (χ2=162.373, P<0.001), but the consistency between the two methods was not significant (Kappa = -0.003, P=0.902). The sensitivity, specificity, ACC, PPV and NPV of mNGS were 91.29%, 28.26%, 81.94%, 87.96%, and 36.11% respectively, compared to corresponding 30.30%, 73.91%, 36.77%, 86.96% and 15.60% of traditional culture respectively. Through analysis, it is confirmed that the sensitivity and specificity between the two methods were statistically significant (91.29% vs 30.30%, χ2=148.120, P<0.001 and 28.26% vs 73.91, χ2=13.793, P<0.001). Conclusions　mNGS can significantly improve the detection rate of pathogens in pulmonary infections and provide a complementary tool besides to traditional culture method for accurate anti-infection therapy. Furthermore, both traditional culture and mNGS pathogen detection methods are highly dependent on sample quality and detection quality control. mNGS requires the correct interpretation of comprehensive, non-destructive pathogenic genetic information to accurately identify pathogens.

Objective To explore the Oneomehnia distribution on electronic map based on gridding orientation pattern of marshlands in Changsha along Xiangjiang River,in order to manage snail information in a visual and digital way.Methods The data of geographic information and snail information were collected by global positioning system(GPS)and database was established using systematic sampling,and then an information management system of electronic map about the distribution of Oncomelania was designed.The degree of Oncomelania was expressed in different colors,the Oncomelania information in the database on the digital map wag reflected accurately using the geographic information system(GIS)technology.Results The electronic map of the distribution information of 20 marshlands in Changsha along Xiangjiang River were drawn.At the same time,the information could be grasped compreheusively.Digit map contains geographic information and database of snail information,such as the row and rnum,the density of alive and infectious snails,the area and geographic information et al.A method of managing and analyzing snail information with higher efficiency was offerred in this article.Conclusion Conventional methods of managing and analyzing Oncomelania information are reformed into digitalization and visualization.

OBJECTIVE To investigate the neuroprotective effects of quercetin on central neurons against chronic high glucose in central neurons, in relation to Nrf2/ARE/Glo-1 activation. METHODS SH- SY5Y cells were cultured with high glucose (HG, 70 mmol · L- 1), 4- fold of the normal glucose (17.5 mmol · L- 1). Quercetin was set three concentrations (5, 10, 20 μmol · L- 1), with Nrf2 activator sulforaphane (SFN) as a positive group (2.5 μmol·L-1). After 72 h, cells were collected for glyoxalase 1 (Glo-1) activity and GSH level were by spectrophotometry; advanced glycation end-products (AGEs) as well as nuclear Nrf2 and p-Nrf2 levels by immunofluorescence; Glo-1, γ-glutamycysteine synthase (γ-GCS), Nrf2 and p-Nrf2 protein levels by Western blotting, and Glo-1 and γ-GCS mRNA levels by real-time qPCR. RESULTS Quercetin increased the cell viability of SH-SY5Y cells, and upregulated the levels of Glo-1 activity, protein, and mRNA in SH-SY5Y cells cultured with HG, accompanied by the elevated levels of glutathione, a cofactor of Glo-1 activity, and the reduced levels of AGEs. Meanwhile, quercetin could increase p- Nrf2 and Nrf2 levels in nucleus as well as p- Nrf2 levels in cytosol of SH-SY5Y cells exposed to chronic HG, accompanied by the elevated protein expression and mRNA levels of γ- GCS, a known target gene of Nrf2/ARE signaling. Moreover, a PKC activator or a p38 MAPK inhibitor pretreatment could significantly increase the protein expression of γ-GCS in HG condition, but an alkylating agent for sulfydryl of cysteine in Keap 1, a negative regulator of Nrf2, pretreatment only showed an increased tendency of γ-GCS protein, compared with without pretreatment; however, after pretreatment with those tool drugs, co-treatment with quercetin and HG had similar results to those of single tool drug pretreatment followed by HG exposure. CONCLUSION Firstly, quercetin can enhance Glo-1 function in central neurons, which is mediated by activation of Nrf2/ARE pathway, then exerts the neuroprotection against HG induced damage; moreover, PKC and p38 MAPK pathways may be involved in Nrf2 inactivation in chronic HG condition.

Objective:To investigate the relationship between the changes in inflammatory markers levels and the onset of post-traumatic stress disorder (PTSD) in the early stage of acute trauma..Methods:From January 2018 to June 2020, patients with acute trauma who were admitted to the Affiliated Hospital of Xuzhou Medical University were selected as subjects. Peripheral venous blood was collected on admission, on the 3rd and 7th day after trauma for routine blood test, C-reactive protein (CRP) and procalcitonin (PCT). The neutrophil to lymphocyte ratio (NLR) was calculated. The PCL-5 scale was used to evaluate PTSD symptoms one month later. The patients were divided into the PTSD group and non-PTSD group with the score of 38 as the boundary. The change rule of NLR in the PTSD group and the non-PTSD group were analyzed.Results:Ninety-one trauma patients were enrolled, including 23 patients in the PTSD group and 68 patients in the non-PTSD group. Compared with the healthy control group, the NLR of 91 trauma patients on admission, on the 3rd and 7th day were significantly higher (all P< 0.01). The NLR of the PTSD group was increased on the 7th day after trauma, which was significantly higher than that of the non-PTSD group ( P= 0.025). The non-PTSD group showed a decreasing trend, of which NLR on the 7th day was significantly lower than that on admission ( P= 0.001). In addition, high level of NLR on the 7th day after trauma (β= 0.206, P= 0.01) was a risk factor for PTSD onset. Conclusions:Dynamic monitoring of the changes in NLR after acute trauma would be of great clinical value to early warning of PTSD.

The data of a child with early-onset epileptic encephalopathy in Qilu Children′s Hospital of Shandong University in February 2020 were analyzed retrospectively.The child was a 4-month-old girl, who was admitted to the hospital because of " repeated convulsions for 4 months and feeding difficulty for 1 month" at the age of 4 months.The patient suffered from epilepsy 1 day after birth, and the epilepsy type was tonic seizures.Severe developmental retardation was observed in the patient.Electroencephalogram showed multifocal discharge, which then turned to hypsarrhythmia.The cranial imaging was negative.Feeding difficulty occurred at the age of 3 months.The genetic testing revealed a de novo heterozygous missense mutation in the FGF12 gene (Arg114His). Various antiepileptic drugs and ketogenic diet were ineffective.There was no attack in 2 months after adding Phenytoin.The child could eat on her own after seizure control, but there was no progress in intellectual and motor development.Mutations in the FGF12 gene lead to poor prognosis of early-onset epileptic encephalopathy, and the seizures are difficult to control.Sodium ion channel blockers such as Phenytoin should be used as soon as possible.

2 mmx3 mm experimental windows were prepared in adamant slippery surfaces of 7 fresh uprooted permanent teeth. The teeth surfaces in the windows area were demineralized to create artificial caries mould of early stage by aciding the experimental teeth surfaces of 0, 12, 24, 48, 72, 96, 120 hours with demineralized liquid (pH 4.5) in vitro. The demineralized changes on the experimental teeth surfaces were detected by dental Optical Coherence Tomography (OCT) system, which were newly developed by our research group recently, and the detecting results were compared with clinical digital photomicrography and scanning electron microscopy on the same tooth sample, to checkout the efficacy and feasibility of dental OCT for early quantification detecting of artificial enamel demineralization in vitro. The dental OCT system can safelydetect early enamel demineralization of micron level and noninvasively obtain fine resolution quantification information both in surfaces view and sectional view; OCT could accurately detect surface demineralization changes on the experimental windows of artificial dental caries as early as after 12 hours aciding treatment, earlier than the visual inspection and clinical digital photomicrography. OCT could obtain both superficial view and sectional view of quantificational demineralization in early enamel caries homeochronously, and had high correlation to the results of ultramicromorphological changes detected by scanning electron microscopy. Dental OCT system developed by our group could accurately detect early artificial dental caries atraumaticly with high sensitivity and safety. Moreover, it can obtain quantification data in micron level without damaging the experimental teeth samples.