Objective: To analysis the pathogenic mutation and the clinical characteristics of a three generation family with congenital pulverulent cataract. Methods: A congenital cataract family was chosen from the First Affiliated Hospital of AnHui Medical University, 5 ml peripheral blood was obtained from each family member to extract genomic DNA.Next generation sequencing was used to detect the mutation in proband (Ⅱ5), Ⅱ6 and Ⅲ8, and Sanger sequencing was applied to verify pathogenic mutation in the whole family members.The mutation site was compared with the gene sequence of 10 000 normal Chinese.PolyPhen-2 and SIFT were applied to analysis the alteration on the protein structure and function and its possible pathogenesis.This study followed the Declaration of Helsinki and was approved by the Ethics Committee of AnHui Medical University (NO.PJ2017-5-17). All patients signed informed consent. Results: The pedigree consisted of 19 members of three generations, including 10 patients and 9 normal family members.Heterozygous mutation of GJA3 gene c. 427G>A (p.G143R) was detected in all patients of the pedigree, but was not found in normal members of the pedigree and 10 000 normal Chinese.The score calculated from SIFT and PolyPhen-2 indicated that the mutation probably had malignant effect on normal protein structure, Swiss-model website analysis showed that the mutation likely altered the secondary structure of the protein CX 46 by reducing an α-helix between 107-115 amino acids.Meanwhile, c.1325-1G>T mutation of HSF4 gene were detected in Ⅱ5 and Ⅲ8, which was not found in other family members and 10 000 normal Chinese.HSF and MaxEntScan results showed that the mutation probably had serious effect on the splicing of mRNA.The cataract development rates of Ⅱ5 and Ⅲ8 were faster than that of the same age in the same generation of the pedigree, and the morphology of lens opacity was changed. Conclusions The heterozygous c. 427G>A mutation in GJA3 gene is responsible for pulverulent cataract in this family, meanwhile the c. 1325-1G>T mutation in HSF4 gene may change the type of phacoscotasmus and accelerate the progress of disease.

Objective:To evaluate the effects of bosutinib on acute lung injury in mice with endotoxemia.Methods:Sixty clean-grade healthy male C57BL/6 mice, aged 8-12 weeks, weighing 20-25 g, were divided into 4 groups ( n=15 each) using a random number table method: control group (group C), bosutinib group (group B), endotoxemia group (group lipopolysaccharide [LPS]) and bosutinib plus endotoxemia group (group B+ LPS). Septic acute lung injury model was developed by intraperitoneal injection of LPS.Bosutinib 5 mg/kg was injected via the tail vein at 0.5 h before establishing the model in group B+ LPS and at the corresponding time point in group B. At 24 h after developing the model, the mice were sacrificed for microscopic examination of the pathological results of lung tissues which were scored for calculation of the lung coefficient (LI) and wet/dry lung weight (W/D) ratio, and for determination of the content of Evans blue in lung tissues (by Evans blue staining), expression of vascular endothelial cadherin (VE-cadherin), vascular cell adhesion molecule 1 (VCAM-1), phosphorylated nuclear transcription factor κB p65 (p-NF-κB p65), phosphorylated nuclear factor κB inhibitory protein α (pIκB-α) (by Western blot) and expression of interleukin-1beta (IL-1β), IL-6 and tumor necrosis factor-alpha (TNF-α) mRNA (using real-time polymerase chain reaction). Results:Compared with group C, the LI, W/D ratio, Evans blue content in lung tissues and lung injury score were significantly increased, and the expression of IL-1β mRNA, TNF-α mRNA, IL-6 mRNA, VCAM-1, p-NF-κB p65 and pIKB-α was up-regulated, and the expression of VE-cadherin was down-regulated in group LPS ( P<0.05), and no significant change was found in the parameters mentioned above in group B ( P>0.05). Compared with group B, the LI, W/D ratio, Evans blue content in lung tissues and lung injury score were significantly decreased, and the expression of IL-1β mRNA, TNF-α mRNA, IL-6 mRNA, VCAM-1, p-NF-κB p65 and pIKB-α was down-regulated, and the expression of VE-cadherin was up-regulated in group LPS ( P<0.05). Conclusions:Bosutinib can ameliorate the acute lung injury in mice with endotoxemia.

Objective:To discuss the mechanism of Yueju Pill in the treatment of functional dyspepsia (FD) based on network pharmacology and molecular docking.Methods:The chemical components and action targets of Yueju Pill were screened out by TCMSP platform and HERB, BATMAN-TCM database combined with literature were used to supplement effective components of Vietnam bow. The targets of FD were screened out by GeneCards database and OMIM database, and the intersection of the two targets was used to analyze the protein interactions using the STRING platform to construct the PPI network. Metascape platform was used for GO and KEGG pathway enrichment analysis, and Cytoscape 3.7.2 software was used to construct a network of "Yueju Pill components-functional dyspepsia targets-pathways". Online mapping tools were used to obtain the Venn plot of the intersection targets of Yueju Pill, FD and its related pathogenesis. Finally, AutoDock software was used for molecular docking.Results:The main active components of Yueju Pill in the treatment of FD are quercetin, wogonin, luteolin, kaempferol, etc. The main targets are AKT1, MAPK1, JUN, RELA, IL6, BCL2, BAX, MAPK8, EGFR, ESR1, etc. Molecular docking shows that the targets and the active components of the Yueju Pill have better binding abilit. The GO enrichment analysis result shows that there are 2 273 biological processes, 152 molecular functions and 91 cell components. KEGG enrichment analysis shows that there are 344 pathways associated with FD. According to literature review, the pathways related to FD include PI3K-Akt signaling pathway, AGE-RAGE signaling pathway, IL-17 signaling pathway, etc.Conclusion:Yueju Pill might act on AKT1, MAPK1, JUN, RELA, IL6, BCL2, BAX, MAPK8, EGFR, ESR1 and other targets to regulate PI3K-Akt signaling pathway, AGE-RAGE signaling pathways and IL-17 signaling pathway and it could treat FD.

OBJECTIVE: To observe the effect of occlusal interference on the afferent pathway of the trigeminal nerve and neuronal excitability in the trigeminal subnucleus caudalis (SPVC) of rats by electrical stimulation of the trigeminal ganglion (TG) and extracellular recordings of SPVC activities. METHODS: Twenty male Wistar rats were randomly divided into control group and model group (=10). In the model group, occlusal interference for 30 consecutive days was induced using light-cured flowable resin on the right maxillary molars. During occlusal interference, the pain sensitivity was scored with von Frey Fibers in the masseter. Simultaneous recordings of electrical activities from the SPVC, electrocardiogram, body temperature and electromyogram of the breath muscles of the anesthetized rats were performed, and the responses evoked by electrical stimulation of the TG were analyzed. RESULTS: Compared with the control rats, the rats in the model group showed significantly increased pain sensitivity scores ( < 0.05) and increased spontaneous discharge frequency of the SPVC ( < 0.05). The amplitude of the SPVC responses induced by electrical stimulation of the TG showed stimulus intensity-dependent changes ( < 0.05), and the amplitude evoked by 4 mA and 8 mA stimulation was similar between the model group and the control group (>0.05). Train stimulation (0.2 ms, 1 mA, 30 s, 100 Hz) of the TG significantly increased the discharge frequency of the SPVC only in the rats in the model group ( < 0.05). CONCLUSIONS The functional activities of the pain afferent pathway of the trigeminal nerve can be electrophysiologically monitored by electrical stimulation of the TG and extracellular recordings of SPVC activities in rats. Occlusal interference can increase the excitability of the neurons in the SPVC and enhance their sensitivities to TG afferent activation, suggesting the neural plasticity of the pain afferent pathway.

ObjectiveTo explore the mechanism of Broussonetiae Fructus （BF） in preventing and treating drug-induced liver injury （DILI） induced by acetaminophen （APAP） through the endoplasmic reticulum stress pathway. MethodSixty C57BL/6N mice were randomly divided into normal group， model group， silybin group （3.4 g·kg^-1）， and high-， medium- and low-dose BF groups （3.0， 1.5， 0.75 g·kg^-1）， with 10 mice in each group. The DILI model was induced by intragastric administration of APAP at 800 mg·kg^-1， and drugs were administered simultaneously for 10 consecutive days. The serum contents or activities of alanine aminotransferase （ALT）， aspartate aminotransferase （AST）， total bilirubin （TBIL）， and direct bilirubin （DBIL） were measured. Hematoxylin-eosin（HE） staining was performed to observe the pathological changes in liver tissues. The morphological changes in liver mitochondria were observed by transmission electron microscopy. The activities or content of superoxide dismutase （SOD）， malondialdehyde （MDA）， total antioxidant capacity （T-AOC）， glutathione （GSH）， glutathione disulfide （GSSG）， glutathione peroxidase （GSH-Px）， and adenosine triphosphate （ATP） in the serum and liver tissues were detected by the colorimetric method. The expression of reactive oxygen species （ROS） in liver tissues was detected by immunofluorescence. The gene expression of glucose-regulated protein 78 （GRP78）， CCAAT/enhancer-binding protein homologous protein （CHOP）， and c-Jun N-terminal kinase （JNK） in liver tissues was detected by Real-time quantitative polymerase chain reaction （PCR）. ResultCompared with the normal group， the model group showed increased serum activities or content of ALT， AST， TBIL， and DBIL （P<0.01）， increased MDA and GSSG contents （P<0.01）， decreased contents or activities of SOD， T-AOC， GSH， GSH-Px， and ATP （P<0.01）， swollen hepatocytes with inflammatory infiltration and lamellar necrosis， swollen and broken mitochondria of hepatocytes， and increased mRNA expression of GRP78， CHOP， and JNK （P<0.01）. Compared with the model group， the groups with drug intervention showed decreased serum content or activities of ALT， AST， TBIL， and DBIL （P<0.05， P<0.01）， reduced MDA and GSSG contents（P<0.05， P<0.01）， and increased contents or activities of SOD， T-AOC， GSH， GSH-Px， and ATP （P<0.05， P<0.01）， improved swollen hepatocytes， inflammatory infiltration， and lamellar necrosis， recovered bilayer membrane structure in mitochondria of hepatocytes， and decreased mRNA expression of GRP78， CHOP， and JNK （P<0.05， P<0.01）. ConclusionBF has preventive and therapeutic effects on APAP-induced DILI mice， and the mechanism may be related to the reduction of endoplasmic reticulum stress and oxidative stress level in vivo.