Objective To set up a method for determining the quercetin in Pyrola calliatha H.Andres.Method HPLC method was carried on Agilent Eclipse XDB-C18 column(4.6 mm?150 mm,5 ?m) with mobile phase of methanol-0.1% phosphoric acid(50:50),and the detecting wavelength was at 370 nm.Result The standard curve was linear within the range of 0.092～0.46 ?g.The average recovery rate was 99.24% and RSD was 0.70%.Conclusion The method is simple,accurate,highly sensitive and reproducible.It can be used for the quality determination of quercetin in Pyrola calliatha H.Andres.

OBJECTIVE To provide the investigative data for timely changing medical gloves according to the study on the dynamic change of physical property of the gloves during abdominal open procedure.METHODS The gloves used in the abdominal open procedures were collected.The tensile strength,break elongation and the morphological values of three types of the gloves(No.6.5,7 and 7.5) were measured and recorded before and after their using at the time point of 0.0h,1.5h,2.5h,3.5h,4.5h,5.5h and 6.5h during operation.RESULTS The tensile strength decreased significantly(P

Objective To observe the tumorigenicity of High Five insect cell line in Balb/c nude mice,and make sure the safety of the cells when used in vaccine production.Methods Balb/c nude mice were randomly divided into 5 groups:the basic cell bank of High Five group,the highest limited passages of High Five group,HEp-2 cell group as positive control,CEF cell group as negative control,and blank control.Except of the blank control,cell suspension was injected subcutaneously into the nude mice in the different groups,respectively.At 3 and 12 weeks after injection,anatomical observation and histopathologic examination were performed to detect the tumor formation.Results At 3 and 12 weeks after injection,the tumorigenicity study results showed that no tumor developed at the transplantation site in the blank control group,negative group,and High Five groups.Histopathological examinations also showed no abnormality in these groups.Otherwise,squamous cell carcinoma was developed in the positive group at 3 weeks after injection.Conclusions High Five cells of basic cell bank and highest limited passages are not tumorigenic,and can be used in vaccine production safely.

Background Retinal microglia (RMG) plays an important role in the pathogenesis of retinal degenerative diseases,while chemokine CX3CL1 participates in the regulation of steady-state of microglia.It has been determined that bone marrow-derived mesenchymal stem cells (BMSCs) have a remarkable role to modulate the immune response and protect the central nervous system through the release of soluble factors in a paracrine fashion and further affect the functional behavior of cells.However,whether BMSCs are able to interact with RMG and activate related signaling pathway for the maintaining of homeostasis in the retina is still unclear.Objective The aim of this study was to investigate the interaction between BMSCs and lipopolysaccharide (LPS)-activated RMG in vitro,and dissect the effects of CX3CL1/CX3CR1 signaling pathway on the biological behavior of BMSCs and RMG.Methods RMG was isolated from SD rats,cultured with mixed culture of retinal glial cells and purified by shaking.The cells were identified by detecting the expression of CD111b,Iba1 and glutamamine synthetase (GS) with indirect immunofluorescence assay.LPS (1 mg/ml,2 μl) was added in the medium for 24 hours to stimulate RMG,and then the cells were divided into LPS control group,BMSCs group (cocultured with BMSCs for 24 hours) and CB-BMSCs group (cocultured with CX3CL1-blocking-BMSCs for 24 hours).The cells without LPS stimulation served as the blank control group.The functions of RMG,including the release content of tumor necrosis factor-α (TNF-α) and interleukin-1 β (IL-1β),the proliferation,phagocytosis,and migration of RMG were examined.Results RMG was successfully isolated and harvested from SD rats by using mixed culture of retinal glial cells and purified by shaking.CD11b and Iba1 showed the positive expression with the green fluorescence in the cells and GS was absent.The contents of TNF-αt in the cell supernatant were (2.55 ±0.97) ng/ml,(24.91 ±3.07) ng/ml,(20.38 ±2.97) ng/ml and (24.90 ± 1.88) ng/ml in the blank control group,LPS control group,BMSCs group and CB-BMSCs group,respectively,showing a significant difference among the groups (F=119.90,P＜0.05).The contents of IL-1 β in the cell supernatant were (1.12±0.36) ng/ml,(10.40±2.76) ng/ml,(7.00± 1.75) ng/ml and (9.55 ± 1.11) ng/ml in the blank control group,LPS control group,BMSCs group and CB-BMSCs group,respectively,showing a significant difference among the groups(F =34.96,P＜0.05).The secretory volume of TNF-α and IL-1 β were evidently lower in the BMSCs group than those in the LPS control group (both at P＜0.05),and no significant differences were found in the secretory volume of TNF-α and IL-1β between CB-BMSCs group and LPS control group (both at P＞0.05).The proliferative rate of RMG was lower in the BMSCs group than that in the LPS control group (P＜0.05),while there was no statistical difference between BMSCs group and CB-BMSCs group (P＞0.05).The mean fluorescence intensity (MFI) and the number of migrated RMG were considerably different among the four groups (F=70.55,15.49,both at P＜0.05),and those in the BMSCs group were significantly increased in comparison with the LPS control group (both at P＜0.05),while there was no significant difference between CB-BMSCs group and LPS control group (both at P＞0.05).Conclusions BMSCs could suppress the proliferation of LPS-activated RMG.Moreover,BMSCs might inhibit proinflammatory cytokines releasing,enhance phagocytosis and migration capabilities of RMG via CX3CL1/CX3CR1 signaling pathway.

To investigate the material foundation and mechanism of Zi Bu Pi Yin Recipe in the prevention and treatment of mild cognitive impairment. The integrated pharmacology platform was used to study the active ingredients, drug targets, and disease targets of 12 Chinese traditional medicines of Zi Bu Pi Yin Recipe, and to construct the target of"Zi Bu Pi Yin Recipe-Chinese herbal medicine-chemical composition-core Target-the key path "of the visual network and network analysis. Results:The platform screened 687 kinds of active ingredients of Zi Bu Pi Yin Recipe including glycosides, sugars and alcohols. and 595 key targets for treating mild cognitive impairment, Including HADHA, HADH, NAD (P) -dependent steroid dehydrogenase-like (NSDHL), and et al. The enrichment analysis of GO and KEGG showed that these key targets were mainly localized in the cytoplasm and mitochondria. The most common molecular functions were ATP binding and protein binding, and participated in purine metabolism, nucleotide metabolism, carbon metabolism pathway, Carbohydrate metabolism and so on. The platform could predict the key target of Zi Bu Pi Yin Recipe in preventing and treating Mild Cognitive Impairment and its related pathways, which lay a good foundation for further revealing its mechanism.

Objective: To observe the clinical effects of sequential treatment with extensive lesion resection, vacuum sealing drainage (VSD) combined with irrigation of oxygen-loaded fluid, and tissue transplantation on hidradenitis suppurativa (HS) in buttocks which recurred after multiple surgeries. Methods: From January 2012 to March 2017, 15 male patients (aged 26-53 years) hospitalized in our burn ward with Hurley′s stage Ⅲ HS in the buttocks recurred after 2-5 operations who met the inclusion criteria were enrolled in the prospective self pre- and post-control study. After extensive resection of the lesion, continuous VSD combined with intermittent irrigation of oxygen-loaded fluid was given, with negative pressure of -16.7 kPa and flow rate of pure oxygen of 1.0 L/min. After 7 days of treatment with VSD combined with irrigation of oxygen-loaded fluid, the negative pressure device was removed and autologous posterior thigh split-thickness skin grafts and/or local flaps were transplanted to repair the wounds. Six patients were performed with split-thickness skin grafting, 4 patients with local flap transplantation, and 5 patients with split-thickness skin grafting together with local flap transplantation. The donor sites of local flaps were sutured directly or transplanted with autologous posterior thigh split-thickness skin grafts, and the donor sites of split-thickness skin grafts with autologous thin split-thickness scalp. The wound tissue or wound granulation tissue was collected before lesion resection and 7 days after treatment with VSD combined with irrigation of oxygen-loaded fluid respectively for bacterial culture and detecting of the content of tumor necrosis factor-alpha (TNF-α) by enzyme-linked immunosorbent assay. Survival of skin grafts and flaps after operation was observed, hospitalization time was recorded, and recurrence and complications of HS were followed up. Data were processed with paired sample t test. Results: Bacterial culture result of wound tissue before lesion resection was positive in all patients, mainly Staphylococcus, Streptococcus, Proteus mirabilis, and anaerobic bacteria, etc., while that of wound granulation tissue after 7 days of treatment with VSD combined with irrigation of oxygen-loaded fluid was negative. The content of TNF-α in wound granulation tissue after 7 days of treatment with VSD combined with irrigation of oxygen-loaded fluid was (10.1±2.9) pg/L, significantly lower than (73.6±5.6) pg/L before lesion resection (t=33.47, P<0.01). The skin grafts and/or flaps of 15 patients survived post operation, and the wounds were healed. The patients were hospitalized for 17-31 days. During follow-up of 1 to 5 years, no recurrence of HS occurred in operative site of buttocks of 15 patients, but 1 patient had ulceration of healed perianal incision caused by high anal fistula and was healed after treatment. Conclusions Sequential treatment with extensive resection, VSD combined with irrigation of oxygen-loaded fluid, and tissue transplantation can thoroughly remove HS lesions in the buttocks and improve the condition of wound bed for skin acceptance after debridement, which is conducive to the cure of HS in the buttocks which has undergone multiple operations but still recurs after operation.

Objective To explore the diagnostic value of Young's modulus obtained by real-time shear wave elastography (SWE) for liver fibrosis in autoimmune hepatitis (AIH) patients. Methods A total of 75 AIH patients in the First Affiliated Hospital of Zhengzhou University from January 2013 to April 2022 were retrospectively enrolled. Scheuer scoring system was used to evaluate degrees of liver fibrosis (S0-S4). By using pathological examination of liver tissues as the golden standard, the receiver operating characteristic curve (ROC) was plotted and the area under the curve (AUC) was used to evaluate the diagnostic value of SWE for the significant fibrosis (≥S2), advanced liver fibrosis (≥S3), and liver cirrhosis (S4), respectively. Independent sample t test was used for comparison of continuous data with normal distribution between the two groups. The Kruskal-Wallis H test was used for comparison of non-normally distributed continuous data between multiple groups and Bonferroni method was used for further comparison between two groups. The Spearman correlation coefficient was used for correlation analysis. The logistic regression analysis was used to predict the impact factors in diagnosis accuracy. Results The Young's modulus measured by SWE was statistically significant different among various fibrosis groups ( H =35.186, P < 0.001) although there was no statistical significance in patients' age and platelet, alanine aminotransferase, aspartate aminotransferase, total bilirubin, alkaline phosphatase, and glutamyl transpeptidase levels (all P > 0.05). The Young's modulus measurement was positively correlated with liver fibrosis ( r =0.675, P < 0.05). The AUCs of SWE in the diagnosis of≥S2, ≥S3, and S4 were 0.839, 0.820 and 0.898, respectively and the corresponding optimum cut-off values were 9.2, 10.9, and 14.4 kPa, respectively. The overall concordance rate of the liver Young' s modulus measurements vs . fibrosis stages was 57.33%. Moreover, the alkaline phosphatase level was an independent predictor for diagnostic accuracy of SWE for stage 0-1 fibrosis ( OR =1.009, 95% CI : 1.001-1.018, P =0.029). Conclusion The SWE possessed a diagnosis value for the significant fibrosis (≥S2), advanced liver fibrosis (≥S3) and liver cirrhosis (S4), although there was a low overall concordance rate in the liver Young's modulus measurements obtained using SWE vs. fibrosis stages.

Tripartite motif (TRIM) family proteins are important effectors of innate immunity against viral infections. Here we identified TRIM35 as a regulator of TRAF3 activation. Deficiency in or inhibition of TRIM35 suppressed the production of type I interferon (IFN) in response to viral infection. Trim35-deficient mice were more susceptible to influenza A virus (IAV) infection than were wild-type mice. TRIM35 promoted the RIG-I-mediated signaling by catalyzing Lys63-linked polyubiquitination of TRAF3 and the subsequent formation of a signaling complex with VISA and TBK1. IAV PB2 polymerase countered the innate antiviral immune response by impeding the Lys63-linked polyubiquitination and activation of TRAF3. TRIM35 mediated Lys48-linked polyubiquitination and proteasomal degradation of IAV PB2, thereby antagonizing its suppression of TRAF3 activation. Our in vitro and in vivo findings thus reveal novel roles of TRIM35, through catalyzing Lys63- or Lys48-linked polyubiquitination, in RIG-I antiviral immunity and mechanism of defense against IAV infection.
A549 Cells ; Animals ; Apoptosis Regulatory Proteins/immunology* ; DEAD Box Protein 58/immunology* ; Dogs ; HEK293 Cells ; Humans ; Influenza A Virus, H1N1 Subtype/immunology* ; Madin Darby Canine Kidney Cells ; Mice ; Mice, Knockout ; Orthomyxoviridae Infections/pathology* ; Proteolysis ; RAW 264.7 Cells ; Signal Transduction/immunology* ; THP-1 Cells ; TNF Receptor-Associated Factor 3/immunology* ; Ubiquitination/immunology* ; Viral Proteins/immunology*

OBJECTIVE: Our previous research showed that Naotaifang (a compound traditional Chinese herbal medicine) extract (NTE) has clinically beneficial effects on neurological improvement of patients with acute cerebral ischemia. In this study, we investigated whether NTE protected acute brain injury in rats and whether its effects on ferroptosis could be linked to the dysfunction of glutathione peroxidase 4 (GPX4) and iron metabolism. METHODS: We established an acute brain injury model of middle cerebral artery occlusion (MCAO) in rats, in which we could observe the accumulation of iron in neurons, as detected by Perl's staining. Using assay kits, we measured expression levels of ferroptosis biomarkers, such as iron, glutathione (GSH), reactive oxygen species (ROS) and malonaldehyde (MDA); further the expression levels of transferrin receptor 1 (TFR1), divalent metal transporter 1 (DMT1), solute carrier family 7 member 11 (SLC7A11) and GPX4 were determined using immunohistochemical analysis, real-time quantitative polymerase chain reaction and Western blot assays. RESULTS: We found that treatment with NTE reduced the expression levels of TFR1 and DMT1, reduced ROS, MDA and iron accumulation and reduced neurobehavioral scores, relative to untreated MCAO rats. Treatment with NTE increased the expression levels of SLC7A11, GPX4 and GSH, and the number of Nissl bodies in the MCAO rats. CONCLUSION Taken together, our data suggest that acute cerebral ischemia induces neuronal ferroptosis and the effects of treating MCAO rats with NTE involved inhibition of ferroptosis through the TFR1/DMT1 and SCL7A11/GPX4 pathways.