[Microglia differential genes and their functions in paraquat-induced Parkinson's disease-like in mice's brains based on single-cell RNA sequencing].

Objective: To analyze the differential genes and related signaling pathways of microglia subpopulations in Parkinson's disease (PD) -like mouse brains induced by paraquat (PQ) based on single-cell RNA sequencing, and provide clues to elucidate the mechanism of PQ-induced PD-like changes in the brain of animals. Methods: In September 2021, six male 6-week-old C57BL/6 mice were randomly divided into control group and experimental group (three mice in each group) . The mice were injected with saline, 10.0 mg/kg PQ intraperitoneally, once every three days, and 10 consecutive injections were used for modeling. After infection, the brains of mice were taken and 10×Genomics single-cell RNA sequencing was performed. Microglia subpopulations were screened based on gene expression characteristics, and Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses were performed. The differential genes of microglia subpopulations between the experimental group and control group were further screened, and functional enrichment analysis was performed using bioinformatics tools. Mouse microglia (BV2 cells) were treated with 0, 60, 90 µmol/L PQ solution, respectively. And real-time fluorescence quantitative PCR experiments were conducted to validate the expressions of differential genes hexokinase 2 (Hk2) , ATPase H+ Transporting V0 Subunit B (Atp6v0b) and Neuregulin 1 (Nrg1) . Results: Cluster 7 and Cluster 20 were identified as microglia subpopulations based on the signature genes inositol polyphosphate-5-phosphatase d, Inpp5d (Inpp5d) and transforming growth factor beta receptor 1 (Tgfbr1) , and they reflected the microglia-activated M2 phenotype. The bioinformatics analysis showed that the characteristic genes of identified microglia subpopulations were enriched in endocytosis. In terms of molecular function, it mainly enriched in transmembrane receptor protein kinase activity and cytokine binding. The up-regulated genes of Cluster 7 were mainly enriched in lysosomal pathway, endocytosis pathway, and down-regulated genes were mainly enriched in neurodegenerative disease and other signaling pathways. The up-regulated genes of Cluster 20 were mainly enriched in signaling pathways related to PD, and down-regulated genes were mainly enriched in cyclic adenosine 3', 5'-monophosphate (cAMP) signaling pathways, neurological development, synaptic function and other signaling pathways. The results of real-time fluorescence quantitative PCR showed that the expressions of Hk2 mRNA and Atp6v0b mRNA increased and the expression of Nrg1 mRNA decreased in the 90 µmol/L PQ-treated BV2 cells compared with the 0 µmol/L, and the differences were statistically significant (P<0.05) . Conclusion: Microglia are activated in the PQ-induced PD-like mouse model and polarized toward the M2 phenotype. And their functions are associated with lysosomal (endocytosis) , synaptic functions and the regulation of PD-related pathways.

[The effection of white matter abnormality to auditory and speech rehabilitation after cochlear implantation in prelingual deafness children].

Objective:To investigatethe effection of white matter abnormality to auditory and speech rehabilitation after cochlear implantation in prelingual deafness children.Method:Thirty-five children with white matter abnormality were included in this study. The degree of leukoaraiosis was evaluated by Scheltens scale based on MRI.The hearing and speechrecovery level was rated by auditory behavior grading standards(CAP) and speech intelligibility grading standards(SIR) at 6 months, 12 months, and 24 months post operation. Result:The CAP scores and SIR scores of the children with white matter abnormality were lower than those of the control group at 6 months after operation (P<0.05).The SIR scores of the children with white matter abnormality at 12 months and 24 months post operation were significantly lower than those of the control group.There was no statistically significant difference between the CAP scores of the two groups at 12 and 24 months after operation(P>0.05).Schelten classification had a greater impact on SIR scores than on CAP scores. Conclusion:The effect of white matter abnormality on auditory and speech rehabilitation after cochlear implantation was related to the degree of leukoencephalopathy. When the lesion of white matter abnormality was larger, the level of hearing and verbal rehabilitation was lower, and the speech rehabilitation was more significantly impacted by white matter lesions degree.

Genetic polymorphisms in the prostaglandin pathway genes and risk of head and neck cancer.

OBJECTIVE: Previous studies examining the association between genetic variations in prostaglandin pathway and risk of head and neck cancer (HNC) have only included polymorphisms in the PTGS2 (COX2) gene. This study investigated the association between genetic polymorphisms of six prostaglandin pathway genes (PGDS, PTGDS, PTGES, PTGIS, PTGS1 and PTGS2), and risk of HNC. METHODS: Interviews regarding the consumption of alcohol, betel quid, and cigarette were conducted with 222 HNC cases and 214 controls. Genotyping was performed for 48 tag and functional single-nucleotide polymorphisms (SNPs). RESULTS: Two tag SNPs of PTGIS showed a significant association with HNC risk [rs522962: log-additive odds ratio (OR) = 1.42, 95% confidence interval (CI): 1.01-1.99 and dominant OR = 1.58, 95% CI: 1.02-2.47; rs6125671: log-additive OR = 1.49, 95% CI: 1.08-2.05 and dominant OR = 1.96, 95% CI: 1.16-3.32]. In addition, a region in PTGIS tagged by rs927068 and rs6019902 was significantly associated with risk of HNC (global P = 0.007). Finally, several SNPs interacted with betel quid and cigarette to influence the risk of HNC. CONCLUSIONS: Genetic variations in prostaglandin pathway genes are associated with risk of HNC and may modify the relationship between use of betel quid or cigarette and development of HNC.

[Experimental study for repair of cranial defects with bone marrow stromal cells and modified alginate].

OBJECTIVE: The primary aim of this investigation was to determine whether expanded BMSCs in vitro mixed with modified alginate gelatin could repair critical defects in rats without the addition of exogenous growth or bone morphogenetic factors. METHODS: Bone marrow stem cells from syngeneic rats cultured in vitro and mixed with modified alginate gel to paint the cranial critical size defect. A full-thickness cranial plate defect was created without damage of dura mater. Modified alginate gelatin with or without BMSCs were painted over the cranial defects. Animals being made cranial defect but received no implant served as sham-operated controls. Craniotomy defects were divided into three groups, which included defects left unpainted (group I, n=6), defects painted with modified alginate gelatin alone (group II, n=6), and defects painted with a modified alginate mixed with BMSCs (group III, n=6). A total of 18 implant experiments were carried out, with postsurgical radiographic and histological analysis completed at 12 week. RESULTS: None of the implants exhibited extrusion or infection. Radiographs showed a likely increased calcification in group III, without finally new calcification in group I and in group II. Histology showed that group I and group II were featured by thinning of the bone at the edges of the defect margins with minimal bone growth inward and dense fibrous tissue with rudimentary alginate material spanning the intervenient gap. The results demonstrated that a great amount of new bone in growth took place in BMSCs-alginate group, stemming from cranial defect edges and proceeding inward. CONCLUSION: Transplantation of syngeneic BMSCs with alginate gel can serve as an example of a cell-based treatment for skeletal reformation and would be especially useful for augmenting or regenerating bone in skeletal defects. So syngeneic BMSCs with alginate gel demonstrate a potential technique to regenerate a variety of skeletal defects that occur in different clinical scenaries.

[Autologous tissue engineered skin for full thickness skin replacement].

OBJECTIVE: To evaluate the feasibility of regenerating or repairing damaged skin utilizing tissue engineering techniques. MATERIALS AND METHODS: Yorkshire pigs were used in the study. A critical size defect of six, full thickness, 4 cm in diameter round wounds, which were marked on the paravertebral region of the animal were excised. The skin defects were randomly divided into three groups. Group I as control group received no cells or polymer. Group II as a second control group received pluronic hydrogel with no cells. Group III as the experimental group received a mixture of cells (keratinocytes and fibroblasts) and pluronic hydrogel. All specimens were harvested at 4 and 6 weeks in vivo, and underwent gross, histological, and transmission electron microscope evaluation. RESULTS: Histologically, the skin in the experimental group was similar to normal skin with stratified epidermis overlying a moderately thick collageneous dermis. The interface of the tissue was apparently demarcated by epidermal ridges and dermal papillae. The control groups showed no skin formation except for granulation with infiltrating inflammatory cells in the wound. Transmission electron microscopy showed that the basal lamina of the experimental group was well developed and attached to the extracellular matrix. CONCLUSION: This finding demonstrates the successful use of tissue engineered skin with Pluronic F-127 as a cell carrier.

[The experimental study of tissue engineered mandible condyle in the shape of human].

OBJECTIVE: This study evaluates the feasibility of creating a tissue engineered adult human mandible condyle composite of bone and cartilage. METHODS: A polymer template formed in the shape of the human mandible condyle using a scaffold composed of polyglycolic acid (PGA) and polylactic acid (PLA) was seeded with osteoblasts isolated from a bovine periosteum suspended in calcium alginate. Chondrocytes isolated from the same calf suspended in 30% pluronic were then "painted" onto the articular surface of the scaffold and implanted into subcutaneous pockets on the dorsa of athymic mice. Animals were divided into three groups: Group I (n=6) received PGA/PLA scaffold saturated with hydrogels not containing cells. Group II (n=6) received scaffolds seeded with both cell types suspended in saline rather than hydrogels. Group III (n=6) received scaffolds seeded with both cell types suspended in hydrogel composites. Constructs were harvested after 12 weeks, and evaluated grossly and microscopically using histologic stains. RESULTS: In group I, constructs formed a small mass without evidence of new bone or cartilage. In group II, constructs were small and irregular in shape. Microscopically they contained scattered islands of bone and cartilage. All specimens in group III retained their original condylar shapes and were quite firm in consistency. Microscopic evaluation revealed trabecular bone interfacing with hyaline cartilage on its articulating surface. CONCLUSION: These findings demonstrate that the composites of bone and cartilage can be engineered to serve as mandibular condylar substitutes. The interdigitation of bone and cartilage at their interface is similar to the normal interface of these composite tissues seen in articulating joints.

Cytotoxic terpenoids from the Formosan soft coral Nephthea brassica.

Two new cytotoxic cembranoid diterpenes, brassicolide (1) and brassicolide acetate (2); a new cytotoxic sesquiterpene, (-)-4alpha-O-acetyl-selin-11-en (3); and six cytotoxic terpenoids, (-)-selin-11-en-4alpha-ol (4), 2-hydroxynephthenol (5), nephthenol (6), cembrene A (7), epoxycembrene A (8), and (-)-beta-elemene (9), have been isolated from the Formosan soft coral Nephthea brassica. The structures of compounds 1-9 were determined by spectral, chemical, and X-ray crystallographic analysis.

[Experimental research on preparation of small root canals with EDTA and cirtric acid]

OBJECTIVE:To study the effects of EDTA and citric acid on small root canals so as to provide experimental basis for clinical applications.METHODS:17% EDTA and 50% citric acid are applied to Group A and Group B respectively for three days, each group having 14 extracted teeth with small root canals. The data obtained during the three days' experiment are analyzed with statistic method.RESULTS:The 14 small root canals applied with 17% EDTA solution for three days are all effective, the overall effectiveness rate being 100% and 11 small root canals show an expansion level of 4mm or over, 9 of which reachthe length required for treatment, 11 out of the 14 small root canals applied with 50% citric acid solution for three days are effective (three are not effective), the overall effectiveness rate being 78.57%. 8 small root canals show an expansion level of over 4 mm and all of them reach the length required for treatment.Based upon statistic treatment, the expansion level of small root canals of both Group A and Group B has an obvious difference (P<0.01) after the application.CONCLUSION:EDTA should be the recommended medicine for chemical preparation of small root canals.

[Application of tricalcium phosphate in one visit root canal treatment].

This paper reports the successful use of tricalcium phosphate (TCP) as periapical barrier material in animal experiments and clinics. The results of animal experiments showed that TCP had good biocompatibility with periapical tissues. At 6 weeks there were osteoids deposited on the surface of TCP. At 12 weeks osteogenesis occurred around the TCP particles and combined each other closely. At 24 weeks around the periapex there were bone deposits and a tendency of apical closure. On the basis of experiments TCP was used to treat 17 chronic periapicitis in 15 patients. All cases were young permanent teeth with immature roots. It is suggested that TCP can be applied directly in chronic periapicitis with open apices of young permanent teeth. From 6 to 12 months after filling with TCP the X-rays showed that radiolucencies reduced or eliminated, bone trabecula formed and periapical lamina dura reestablished.

[Studies on ecology of Fasciolopsis buski and control strategy of fasciolopsiasis].

The development of eggs of Fasciolopsis buski requires oxygen and the eggs cannot survive anaerobic conditions. The eggs have some resistance to low temperature and can be maintained at 4 degrees C for 3 to 4 months; however, the eggs are killed at 50 degrees C in four hours. The presence of salts can influence the development time of the eggs and reduce their hatching rate. Encysted cercariae exist not only on aquatic plants, but also on the surface of the water. The number of encysted cercariae floating on the water surface is about 3.6% of that of the total encysted cercariae. By inquiring into the case history we found that 10.3-12.8% of the patients and 35.1-40% of the infested pigs were possibly infected by drinking water contaminated with encysted cercariae. The authors suggest the use of fermented silage to feed pigs instead of fresh aquatic green fodders to prevent infection in the animals. In addition, aquatic plants such as water chestnut should be boiled for 1 to 2 minutes before eating to kill the encysted cercariae on the plants.