Objective To observe the toxicity of fangyouling after one month’s transdermal administration in rabbits and evaluate its security. Methods Forty rabbits were randomly divided into 4 groups including a control group and low,middle and high dose groups of fangyouling. The rabbits in the control group were administered with sunflower oil,and the other rabbits were administrated dermally with fangyouling of 50,300 and 2 000 mg/kg respectively once a day for 4 weeks. The general con?dition,the skin irritation reaction,body weight,food consumption,hematology,blood biochemistry,organ coefficients and his?topathological changes of all the rabbits were observed. Results There was no obvious effect on the general condition in all the rabbits. However,the mild skin irritation was observed in 2 rabbits of the middle dose group and 4 rabbits of the high?dose group. The decreases of body weight and food consumption were noted in the high dose group. No changes were detected of hema?tology,blood biochemistry or viscera pathological at all dose levels. Conclusion The dose of non?toxic response of fangyouling is 50 mg/kg at this study condition.

Objective To investigate the effect of sevoflurane anaesthesiaon the expression of N-methyl-D-aspartate (NMDA ) receptor in the developing hippocampal neurons in neonatal rats .Methods Sixty-four healthy male Sprague-Dawley rats ,aged 7 days ,weighing 10-15 g ,were randomly divided into 2 groups with 32 rats in each group using a random number table:control group (group C ) and sevoflurane anaesthesia group (group S ) . Animals in group C inhaled 30% oxygen for 6 h ,while animals in group S inhaled 3% sevoflurane for 6 h .Y-maze test was performed in the rats at 21 and 28 days after birth to evaluate the memory function .On 7 days after birth (immediately after the end of oxygen inhalation or sevoflurane anesthesia ) ,and 14 ,21 and 28 days after birth ,the expression of 1 ,2A and 2B subunits-containing NMDA receptors in the total protein and membrane protein in hippocampal neurons was determined by Western blot .The ratio of NMDA receptors in the membrane protein to those in the total protein (m/t ratio ) was calculated .Results Compared with group C ,the percentage of spontaneous alternation was significantly decreased on 21 and 28 days after birth ,the expression of 1 ,2A and 2B subunits-containing NMDA receptors in the membrane protein was down-regulated on 7-28 days after birth ,and m/t ratio was decreased in group S ( P<0.05) .There was no significant difference in the number of entries into each arm and expression of 1 ,2A and 2B subunits-containing NMDA receptors in the total protein between group C and group S ( P>0.05 ) .Conclusion The mechaism by which sevoflurane anaesthesia induces memory impairment in neonatal rats is related to inhibition of trafficking of NMDA receptors in the developing hippocampal neurons to the cell membrane ,and down-regulation of the number of NMDA receptors in the membrane protein .

Objective To investigate the clinical features, prognosis and risk factors of patients of acquired immunodeficiency syndrome (AIDS) complicated with cryptococcal meningitis (CM). Methods Totally 26 patients of AIDS with CM who were hospitalized in the No. 8th People's Hospital of Guangzhou were enrolled in this study. The clinical data including diagnosis,experimental and etiological test,treatments and prognosis from all the patients were analyzed retrospectively. The results of cerebrospinal fluid routine test and CD4+ T lymphocyte were compared with those of AIDS patients complicated with tuberculous meningitis. Results Among the 26 patients enrolled in the study, the positive rate of cerebrospinal fluid india ink smear or Crypotococcus neoforrnans euhure was 84.6%. The most common symptoms were fever, headache and meningeal irritation sign. The average CD4+ lymphocyte count was 17.83 × 106/L, which was statistically different from that of tuberculous meningitis patients. All the patients showed concomitant multiple organ infections. The mortality rate was as high as 42.3%. At the end of therapy, the cell counts in the eerebrospinal fluid were remarkably higher in the patients with unfavorable prognosis compared to the patients with good prognosis, which was statistically different. Conclusions CD4+ lymphocyte count is an important marker for differentiating CM from tuberculous meningitis in AIDS patients. The results of cerebrospinal fluid routine test can predict the prognosis.

Biomedical Research ; Clustered Regularly Interspaced Short Palindromic Repeats ; Humans

Objective To investigate the role of hippocampal α7 nicotinic acetylcholine receptor (α7nAChR) in sevoflurane-induced deficit in long-term cognitive function in neonatal rats.Methods Sixty-four male Sprague-Dawley rats,aged 7 days,weighing 10-15 g,were randomly divided into 4 groups (n =16each) using a random number table:control group (group C),sevoflurane anesthesia group (group S),sevoflurane anesthesia + α7nAChR agonist PNU-282987 group (group PS),and α7nAChR inhibitor MLA group (group M).In C and S groups,the rats inhaled 30％ oxygen and 3％ sevoflurane for 6 h,respectively.In group PS,PNU282987 (5 mg/kg) was injected intraperitoneally and 24 h later the rats were exposed to 3％ sevoflurane for 6 h.In group M,MLA 3 mg/kg was injected intrappritoneally and 24 h later the rats inhaled 30％ oxygen for 6 h.Eight rats in each group were randomly chosen and sacrificed immediately after oxygen or sevoflurane inhalation.The hippocampus was renoved for determination of the expression of α7nAChR and NR1,NR2A and NR2B subunitscontaining NMDA receptors in the total protein and membrane protein in hippocampal neurons.When the left rats in each group were raised to 2 months,Y-maze test was performed to detect the cognitive function.Results Compared with group C,the expression of α7nAChR and NR1,NR2A and NR2B subunits-containing NMDA receptors in the membrane protein was significantly down-regulated,and the percentage of spontaneous alternation was decreased in group S,the expression of NRI and NR2A subunits-containing NMDA receptors in the membrane protein was down-regulated (P ＜ 0.05),and no significant change was found in the expression of NR2B subunitscontaining NMDA receptors in the membrane protein and percentage of spontaneous alternation in group PS (P ＞ 0.05),and no significant change was found in the expression of NR1 and NR2A subunits-containing NMDA receptors in the membrane protein (P ＞ 0.05),and the expression of NR2B subunits-containing NMDA receptors in the membrane protein was down-regulated,and the percentage of spontaneous alternation was decreased in group M (P ＜ 0.05).Compared with group S,no significant change was found in the expression of NR1 and NR2A subunits-containing NMDA receptors in the membrane protein (P ＞ 0.05),and the expression of NR2B subunitscontaining NMDA receptors in the membrane protein was significantly up-regulated,and the percentage of spontaneous alternation was increased in PS group (P ＜ 0.05).There was no significant difference in the expression of α7nAChR and NR1,NR2A and NR2B subunits-containing NMDA receptors in the total protein and the number of entries into each arm in Y-maze test between the four groups (P ＞ 0.05).Conclusion The mechanism by which sevoflurane induces deficit in long-term cognitive function may be related to decreased function of hippocampal α7nAChR and inhibition of function of NMDA receptors in neonatal rats.

Objective To investigate the anatomical structure of the first plantar lumbrical muscle in the foot and to measure the relevant data which could provide anatomical basis for repairing thumb and finger defects with the transplantation of toes accompanied with the first lumbrical muscle,and to explore the marphological function of the first lumbrical muscle of the foot.Methods From March,2016 to January,2018,a systematic and detailed dissection of the 50 formalin-fixed feet was performed to observe the exact position of the starting and ending points of the first lumbrical muscle,and a Vernier caloper was used to measure the relevant record data.Results The first lumbrical muscle originates from the medial portion of the flexor digitorum lungus tendon of the second toe,and the length of the ventral muscle was [55.87±8.67(79.30-41.16] mm.There were 2 endpoints in the tendon.The first one was in the medial tubercle of the proximal phalanges.The second one was aponeurosis of the dorsal toe and the tendon was divided into proximal and distal segments with the medial tubercle as the mark point.The length of the proximal segment was [15.34±4.81(5.52-25.18] mm,the width of the proximal segment was [2.31±1.12(3.28-1.21)] mm,the thickness was [0.44±0.14(0.28-0.68)] mm;the length of the distal segment was [11.51±4.06(3.46-14.90)] mm,the width was [6.10±1.44(9.36-3.70)] mm,and the thickness was [0.18±0.09(1.10-0.38)] mm.The length and thickness of the proximal segment was signifantly larger than those of the distal segment (P＜0.05).Conclusion The first lumbrical muscle has the function of maintaining the balance and stability of both the toe and the arch during movement,flexuring the metatarsophalangeal joint,extending the interosseous joint of the extensor phalangeal,adducting the second toe;also the function of preventing the second toe from pronation during foots' movement.

PURPOSE: Hrd1 has recently emerged as a critical regulator of B-cells in autoimmune diseases. However, its role in the pathogenesis of chronic rhinosinusitis with nasal polyps (CRSwNP) remains largely unexplored. This study aimed to examine Hrd1 expression and B-cell accumulation and their possible roles in CRSwNP. METHODS: Quantitative real-time polymerase chain reaction, immunohistochemistry, enzyme-linked immunosorbent assay and Western blotting were used to assess gene and protein expression in nasal tissue extracts. Cells isolated from nasal tissues and peripheral blood mononuclear cells were characterized by flow cytometry. Local antibody production was measured in tissue extracts with a Bio-Plex assay. Additionally, changes in Hrd1 expression in response to specific inflammatory stimuli were measured in cultured dispersed polyp cells. RESULTS: Nasal polyps (NPs) from patients with eosinophilic CRSwNP (ECRS) had increased levels of Hrd1, B-cells and plasma cells compared with NPs from patients with non-eosinophilic CRSwNP (non-ECRS) or other control subjects (P < 0.05). The average Hrd1 levels in B-cells in NPs from ECRS patients were significantly higher than those from non-ECRS patients and control subjects (P < 0.05). NPs also contained significantly increased levels of several antibody isotypes compared with normal controls (P < 0.05). Interestingly, Hrd1 expression in cultured polyp cells from ECRS patients, but not non-ECRS patients, was significantly increased by interleukin-1β, lipopolysaccharide and Poly(I:C) stimulation, and inhibited by dexamethasone treatment (P < 0.05). CONCLUSIONS: Differential Hrd1 expression and B-cell accumulation between the ECRS and non-ECRS subsets suggests that they can exhibit distinct pathogenic mechanisms and play important roles in NP.
Antibody Formation ; Autoimmune Diseases ; B-Lymphocytes* ; Blotting, Western ; Dexamethasone ; Enzyme-Linked Immunosorbent Assay ; Eosinophils* ; Flow Cytometry ; Humans ; Immunity, Innate ; Immunohistochemistry ; Nasal Polyps* ; Plasma Cells ; Polyps ; Real-Time Polymerase Chain Reaction ; Tissue Extracts

PURPOSE: Breast cancer is the most frequently diagnosed malignancy in women worldwide. MicroRNAs (miRNAs) are thought to serve as potential biomarkers in various cancers, including breast cancer. METHODS: We evaluated the miRNA expression profiles in 1,083 breast cancer samples and 104 normal breast tissues from The Cancer Genome Atlas database. We used the edgeR package of R software to analyze the differentially expressed miRNAs in normal and cancer tissues, and screened survival-related miRNAs by Kaplan-Meier analysis. A receiver operating characteristic curve was generated to evaluate the accuracy of these miRNAs as molecular markers for breast cancer diagnosis. Furthermore, the functional role of these miRNAs was verified using cell experiments. Targets of candidate miRNAs were predicted using 9 online databases, and Gene Ontology (GO) functional annotation and pathway analyses were conducted using Database for Annotation, Visualization and Integrated Discovery online tool. RESULTS: A total of 68 miRNAs showed significantly different expression patterns between the groups (p < 0.001), and 13 of these miRNAs were significantly associated with poor survival (p < 0.05). Three miRNAs with high specificity and sensitivity, namely, miR-148b-3p, miR-190b, and miR-429, were selected. In vitro experiments showed that the overexpression of these 3 miRNAs significantly promoted the proliferation and migration of MDA-MB-468 and T47D cells and reduced the apoptosis of T47D cells. GO and pathway enrichment analyses revealed that the targets of these dysregulated miRNAs were involved in many critical cancer-related biological processes and pathways. CONCLUSION: The miR-148b-3p, miR-190b, and miR-429 may serve as potential diagnostic and prognostic markers for breast cancer. This study demonstrated the roles of these 3 miRNAs in the initiation and progression of breast cancer.
Apoptosis ; Biological Phenomena ; Biological Processes ; Biomarkers ; Breast Neoplasms ; Breast ; Diagnosis ; Female ; Gene Ontology ; Genome ; Humans ; In Vitro Techniques ; Kaplan-Meier Estimate ; MicroRNAs ; ROC Curve ; Sensitivity and Specificity

Objective:To examine the clinical experience and efficacy of unrelated cord blood transplantation (UCBT) in the treatment of recurrent refractory Epstein-Barr virus-associated hemophagocytic lymphohistiocytosis (EBV-HLH) in children.Methods:The clinical data of a patient with recurrent refractory EBV-HLH and intestinal perforation who was treated by UCBT in Department of Pediatrics, the First Affiliated Hospital of Zhengzhou University in September 2015 and finally cured were retrospectively analyzed.Meanwhile, literature was reviewed.Results:The patient, male, 1 year and 6 months, was admitted to the hospital with " fever for 15 days, rash for 9 days" as the main complaint, mainly manifested as high fever, large liver, spleen, lymph nodes, rapidly progressing pancytopenia, liver function damage, phagocytic blood cells on bone marrow smear, diagnosed as EBV-HLH in September 2015.The patient received chemotherapy according to the HLH-2004 protocol developed by the International Association of Cell Societies.During the treatment, he suffered two recurrence during the maintenance period, and a second-line rescue treatment was adopted, namely, " Pegaspargase, Doxorubicin liposome, Etoposide and Methylprednisolone" (L-DEP regimen) chemotherapy.The complete relief of diagnostic indexes for hemophagocytic lymphohistiocytosis was evaluated after chemotherapy.The patient developed sudden intestinal perforation and underwent emergency surgical surgery, enteroenterostomy.After the condition was stabilized, the patient was pretreated with the " Fludarabine+ Busulfan+ Cyclophosphamide" (Flu+ BU+ CY) therapy and then treated with UCBT, with intravenous nutritional support provided during the entire process.Neutrophil and platelet implantation was implemented on day 13 and day 35 after transplantation, respectively.The chimeric rate was 100%, and the implantation was a success.Hepatic veno-occlusive disease, fungal pneumonia and skin graft-versus-host disease (GVHD) Ⅱ occurred on the 15 th day, 22 nd day and 26 th day after transplantation, respectively.The corresponding symptoms improved after treatment.On day 49 after transplantation, phase Ⅱ " enterostomy fistula" was performed.The patient was followed up to 70 months after transplantation, and generally in good condition.His symptoms relieved, and no chronic GVHD and other comorbidities occurred. Conclusions:Allogeneic hematopoietic stem cell transplantation is the only possible effective means of treating relapsed refractory EBV-HLH in children.In the absence of a suitable sibling or unrelated donor, unrelated cord blood stem cells can be used as a graft source.Enterostomy after intestinal perforation is not contraindicated for transplantation.

Objective:To investigate the clinical value of the expression levels of biological protein markers regenerating islet-derived protein 3-alpha(REG3α), soluble tumor suppressor factor 2(sST2) and tumor necrosis factor receptor 1(TNFR1) in peripheral blood in the diagnosis and efficacy evaluation of intestinal acute graft-versus-host disease (aGVHD) in children after allogeneic hematopoietic stem cell transplantation (allo-HSCT).Methods:Retrospective analysis of 50 children who underwent allo-HSCT, in the Department of Pediatrics, the First Affiliated Hospital of Zhengzhou University from January 2020 to February 2021 were enrolled, including 39 males and 11 females [median age: 8.5 (1-13) years]. The expression levels of above 3 biological proteins were detected before transplantation, 1 week, 2 weeks, 3 weeks, 5 weeks, 7 weeks, 9 weeks, 11 weeks and 13 weeks after transplantation, when intestinal aGVHD occured, and after treatment.Children with intestinal aGVHD were taken as the observation group, and children without intestinal aGVHD were taken as the control group.Whether differences in the expression levels of the 3 biological proteins in the peripheral blood of the 2 groups of children were statistically significant was analyzed.The receiver operating characteristic(ROC) curve was used to evaluate the diagnostic value of the above three biological proteins for intestinal aGVHD, and independent sample t test was performed to compare the expression levels of the 3 biological proteins before and after treatment in children with intestinal aGVHD. Results:(1) The concentrations of REG3α, sST2, and TNFR1 in the peripheral blood of the observation group were (33 985.42±24 631.33) ng/L, (139 899.66±115 825.65) ng/L, (3 041.65±2 418.72) ng/L, respectively, which were higher than the control group of (7 457.39±4 547.49) ng/L, (32 059.57±23 452.85) ng/L, (1 944.51±1 170.35) ng/L, the difference was statistically significant ( t=6.04, 5.19, 2.17, all P<0.05). (2) The area under ROC curve (AUC) of REG3α combined with sST2 in the diagnosis of intestinal aGVHD was 0.952 (95% CI: 0.851-0.992, P<0.001), the maximum Youden index was 0.894, the corresponding sensitivity was 83%, and the specificity was 99%.Its diagnostic value was better than REG3α, sST2 and TNFR1 ( Z=1.763, 1.332, 3.001, all P<0.05). (3) The concentrations of REG3α, sST2, and TNFR1 before treatment in the peripheral blood of children having received effective treatment were (31 343.01±25 364.71) ng/L, (146 629.52±110 501.04) ng/L and (2 489.00±859.70) ng/L, respectively, which were (12 104.37±11 704.60) ng/L, (93 539.55±81 920.93) ng/L and (2 048.15±813.47) ng/L after treatment, lower than those before treatment.The expression levels of REG3α and sST2 were significantly reduced ( t=-3.23, -2.10, all P<0.05), while the difference of the expression level of TNFR1 before and after treatment was not statistically significant ( P>0.05). Conclusions:REG3α and sST2 can be used as important reference indicators for clinical auxiliary diagnosis of intestinal aGVHD, and have good auxiliary diagnostic value.REG3α and sST2 can be used as objective indicators to evaluate the efficacy of clinical treatment of intestinal aGVHD.