To explore the roles of astrocytes in the epileptogenesis, astrocytes and neurons were isolated, purified and cultured in vitro from cerebral cortex of rats. The astrocytes were activated by ciliary neurotrophic factor (CNTF) and astrocytic conditioned medium (ACM) was collected to treat neurons for 4, 8 and 12 h. By using Western blot, the expression of calmodulin dependent protein kinase Ⅱ (CaMK Ⅱ), inducible nitric oxide synthase (iNOS) and adenylate cyclase (AC) was detected in neurons. The results showed that the expression of CaMK Ⅱ, iNOS and AC was increased significantly in the neurons treated with ACM from 4 h to 12 h (P＜0.05), and that of iNOS and AC peaked at 8 h and 12 h respectively. It was suggested that there might be some epileptogenic factors in the ACM and such signal pathways as NOS-NO-cGMP, Ca2+/CaM-CaMK Ⅱ and AC-cAMP-PKA might take part in the signal transduction of epileptogenesis.

BACKGROUND:Vacuum sealing drainage is currently considered as a safe and effective for fasciotomy wounds in the treatment of compartment syndrome. But the wounds after treatment are often not self-closed, which needs skin grafts that can cause secondary injury. Studies have shown that shoelaces technology is useful for leg fasciotomy wounds in the surgical treatment of lower leg compartment syndrome, but so far there are few domestic reports. OBJECTIVE:To compare the vacuum sealing drainage and shoelace technique for treatment of leg fasciotomy wounds. METHODS:A total of 36 patients, with 46 leg fasciotomy wounds, were randomized into vacuum sealing drainage and shoelace technique groups, 23 wounds in each group. In the vacuum sealing drainage group, patients were subjected to vacuum sealing drainage after fasciotomy based on fracture reduction and external fixation;in the shoelace technique group, the fasciotomy wounds were covered with polyethylene/ethanol hydrated seaweed salt foam fol owed by shoelace technique. After 1 month, wound size, wound closure time, infection, further intervention and daily treatment cost were compared between the two groups. RESULTS AND CONCLUSION: Wound closure time was significantly higher in the vacuum sealing drainage group compared to the shoelace technique group (P < 0.05). Eight wounds in the vacuum sealing drainage group required skin grafts; while, no skin graft was necessary in the shoelace technique group. In the shoelace technique group, the vessel loops had to be replaced in five wounds. There was no wound infection, increased compartment pressure and skin flap necrosis postoperatively in both two groups. Both vacuum sealing drainage and the shoelace technique are safe, reliable and effective methods for closure of leg fasciotomy wounds. Vacuum sealing drainage requires longer time to definite wound closure and is far more expensive than the shoelace technique, especially when additional skin grafting is required.

Objective To study the stability of the toxicity,antigenicity and immunogenicity of Neisseria meningitidis serogroup A/C[CMCC(B)29201/29205],and to analyze the quality of the capsular polysaecharide extracted from Neisseria meningitidis.Methods The toxicity of the 3rd,5th,10th,15th,20th,25th and 30th passage of the Neisseria meningitidis was assayed in mice,and the antigenicity of each passage was measured by the tube agglutination test(TAT)and ELISA.The effect of individual 30 passages of Neisseria nveningitidis on the brain tissue and the immunogenicity of each passage were tested in mice,and the capsular polysaccharide was purified and analyzed.Resuits The LD50 of the strains CMCC(B)29201/29205 of each passage was low(≥109/ml),and all the 30 passages of the two strains had no effect on the brain tissue of the mice.The titer of each passage was 1∶320 in TAT and more than 1∶3752 in ELISA.After immunization with individual 30 passages of the Neisseria meningitidis the titers in serum bactericidal assay (SBA)were all more than 1∶32.The capsular polysaccharide purified from Neisseria meningitidis met the quality standard of the requirement.Conclusion The strains of Neisseria meningitidis serogroup A/C used in the manufacture of the meningococcal conjugate vaccine,are stable in the toxicity,antigenicity and immunogenicity.And the capsular polysaccharide has met the quality standard.

Objective: To investigate the effect of salivary adenoid cystic carcinoma (SACC) derived exosomes on PD-L1 expression in fibroblasts. Methods: Exosomes of SACC-83 cell line were extracted by exosome isolation kit, and its particle size, density and phenotypes were identified by electron microscope.After being labeled with PKH67 fluorescence, exosomes were co-incubated with HPLF to observe whether the exosomes could be ingested by fibroblasts under confocal microscope. After co-incubation with the exosomes, the differentially expressed genes (DEGs) in HPLF cells were detected by whole transcriptome sequencing. GO analysis together with KEGG enrichment analysis was used to clarify the biological functions and related signaling pathways related with the DEGs. qPCR, Western blotting and Flow cytometry were used to detect the effect of tumor exosomes on the mRNAand protein expressions of PD-L1, LAG3 and IDO1 in HPLF fibroblasts. Results: SACC exosomes specifically expressed CD63, CD81 and TSG101 molecules, and could be ingested by fibroblasts. After the treatment of fibroblasts by exosomes, the expression of PD-L1 molecule was significantly up-regulated (Fold change=10.19), and the DEGs were significantly enriched in the immune response signaling pathways, such as TNF, NF-κB and cGAS-String pathway, etc. In vitro experiments showed that exosomes could significantly promote the expression of PD-L1 in HPLF cells at both mRNA and protein levels (24.7±4.75 vs 1.03±0.11,P<0.05). Conclusion: Exosomes of SACC can promote the expression of immunocheckpoint ligand PD-L1 in fibroblasts.

Objective To explore and analyze the clinical effect of enhanced recovery after surgery to tibial plateau fractures patients applied arthroscopic minimally invasive treatment. Methods A total of 60 tibial plateau fractures patients were selected in our orthopedics department from January 2016 to July 2017 and who applied arthroscopic minimally invasive treatment, according to the last two-digit number of patient ID, divided them into observation group (n=30) and control group (n=30) randomly. The control group used regular perioperative strategies. The observation group used multidisciplinary cooperation fast track surgery idea, through preoperative assessment and education, nutrition and fasting, advance pre-rehabilitation and preventive analgesia; intraoperative optimization of anesthesia, body fluid management and body temperature control; postoperative nutritional support, multimodal analgesia, early ambulation and rehabilitation exercises, implied standardized and professional perioperative overall optimization management. The differences of the condition of 6 h, 12 h, 24 h after surgery, VAS score on discharge, time of first ambulation, active knee flexion 120°days; self-care ability at discharge and AKSS score one month after surgery between 2 groups were compared. Results The VAS scores 6 h, 12 h, 24 h after surgery and at discharge were 4.48 ± 1.18, 3.81 ± 1.68, 3.05 ± 1.63, 2.65 ± 1.65 in the observation group, and were 5.45±1.15, 4.15±1.05, 3.71±1.15, 3.23±1.68 in the control group. The differences were statistically significant (t=0.796~0.902 , P<0.05). The time of first ambulation, active knee flexion 120° days, self-care ability at discharge and AKSS scores one month after surgery were (5.61±1.4) hours, (4.01± 1.1) days, 80.22±3.6, 71.89±6.56 and 64.13±6.15 in the observation group, and (35.8±8.1) hours, (6.82± 1.6) days, 64.25±3.8, 63.45±8.36 and 60.95±8.98 in the control group. The differences were statistically significant (t=2.789~10.200, P<0.05). Conclusion Enhanced recovery after surgery to tibial plateau fractures patients applied arthroscopic minimally invasive treatment is worthy of being popularized as it’s beneficial to tibial plateau fractures patients. It can also fasten recovery and improve quality of life for postoperative patients.

In this study, apoptosis was induced by new type gosling viral enteritis virus (NGVEV) in experimentally infected goslings is reported in detail for the first time. After 3-day-old goslings were orally inoculated with a NGVEV-CN strain suspension, the time course of NGVEV effects on apoptotic morphological changes of the internal tissues was evaluated. These changes were observed by histological analysis with light microscopy and ultrastructural analysis with transmission electron microscopy. DNA fragmentation was assessed with a terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) assay and DNA ladder analysis. A series of characteristic apoptotic morphological changes including chromatin condensation and margination, cytoplasmic shrinkage, plasma membrane blebbing, and formation of apoptotic bodies were noted. Apoptosis was readily observed in the lymphoid and gastrointestinal organs, and sporadically occurred in other organs after 3 days post-infection (PI). The presence and quantity of TUNEL-positive cells increased with infection time until 9 days PI. DNA extracted from the NGVEV-infected gosling cells displayed characteristic 180~200 bp ladders. Apoptotic cells were ubiquitously distributed, especially among lymphocytes, macrophages, monocytes, and epithelial and intestinal cells. Necrosis was subsequently detected during the late NGVEV-infection phase, which was characterized by cell swelling, plasma membrane collapse, and rapidly lysis. Our results suggested that apoptosis may play an important role in the pathogenesis of NGVE disease.
*Adenoviridae/classification/pathogenicity ; Adenoviridae Infections/pathology/*veterinary/virology ; Animals ; *Anseriformes ; *Apoptosis ; Bird Diseases/*virology ; DNA Fragmentation ; Enteritis/*veterinary/virology ; Epithelial Cells/cytology/virology ; In Situ Nick-End Labeling ; Intestines/cytology/virology ; Leukocytes/cytology/virology ; Lymphoid Tissue/cytology/virology ; Macrophages ; Microscopy, Electron, Transmission

Objective:To investigate the safety and efficacy of haplo-identical hematopoietic stem cell transplantation (haplo-HSCT) conditioning with the same dosage form of antithymoglobulin (ATG) in patients with severe aplastic anemia (SAA) failure to ATG.Methods:This was a retrospective cohort study. A total of 65 patients with SAA who failed ATG treatment and received haplo-HSCT conditioning with the same dosage of ATG at the Institute of Hematology, Peking University People′s Hospital between July 2008 and October 2020 were included as the ATG treatment failure group. An additional 65 SAA patients who applied ATG for the first time during haplo-HSCT were randomly selected by stratified sampling as the first-line haplo-HSCT group. Baseline clinical data and follow-up data of the two groups were collected. Conditioning-related toxicity within 10 days after ATG application and long-term prognosis were analyzed. The Kaplan-Meier was used to calculate the overall survival rate, and the Log-rank test was applied to compare the rates of the two groups.Results:In the ATG treatment failure group, there were 36 males and 29 females, and the age at the time of transplantation [ M ( Q1, Q3)] was 16 (8, 25) years. In the first-line haplo-HSCT group, there were 35 males and 30 females, with a median age of 17 (7, 26) years. Within 10 days of ATG application, the incidence of noninfectious fever, noninfectious diarrhea, and liver injury in the ATG treatment failure group was 78% (51 cases), 45% (29 cases), and 28% (18 cases), respectively, and in the first-line haplo-HSCT group was 74% (48 cases), 54% (35 cases), and 25% (16 cases), respectively; the difference between the two groups was not statistically significant for any of these three parameters (all P>0.05). For graft-versus-host disease (GVHD), there was no significant difference between the ATG treatment failure group and the first-line haplo-HSCT group in the development of 100 day Ⅱ to Ⅳ acute GVHD (29.51%±0.35% vs. 25.42%±0.33%), Ⅲ to Ⅳ acute GVHD (6.56%±0.10% vs. 6.78%±0.11%), and 3-year chronic GVHD (26.73%±0.36% vs. 21.15%±0.30%) (all P>0.05). Three-year overall survival (79.6%±5.1% vs. 84.6%±4.5%) and 3-year failure-free survival (79.6%±5.1% vs. 81.5%±4.8%) were also comparable between these two groups (both P>0.05). Conclusions:Compared with no exposure to ATG before HSCT, similar early adverse effects and comparable survival outcomes were achieved in patients with SAA who failed previous ATG treatment and received haplo-HSCT conditioning with the same dosage form of ATG. This might indicate that previous failure of ATG treatment does not significantly impact the efficacy and safety of salvaging haplo-HSCT in patients with SAA.

Background The benchmark dose (BMD) method calculates the dose associated with a specific change in response based on a specific dose-response relationship. Compared with the traditional no observed adverse effect level (NOAEL) method, the BMD method has many advantages, and the 95% lower confidence limit of benchmark dose lower limit (BMDL) is recommended to replace NOAEL in deriving biological exposure limits. No authority has yet published any health-based guideline for rare earth elements. Objective To evaluate genotoxicity threshold induced by acute exposure to neodymium nitrate in mice using BMD modeling through micronucleus test and comet assay. Methods SPF grade mice (n=90) were randomly divided into nine groups, including seven neodymium nitrate exposure groups, one control group (distilled water), and one positive control group (200 mg·kg⁻¹ ethyl methanesulfonate), 10 mice in each group, half male and half female. The seven dose groups were fed by gavage with different concentrations of neodymium nitrate solution (male: 14, 27, 39, 55, 77, 109, and 219 mg·kg⁻¹; female: 24, 49, 69, 97, 138, 195, and 389 mg·kg⁻¹) twice at an interval of 21 h. Three hours after the last exposure, the animals were neutralized by cervical dislocation. The bone marrow of mice femur was taken to calculate the micronucleus rate of bone marrow cells, and the liver and stomach were taken for comet test. Results The best fitting models for the increase of polychromatophil micronucleus rate in bone marrow of female and male mice induced by neodymium nitrate were the exponential 4 model and the hill model, respectively. The BMD and the BMDL of female mice were calculated to be 31.37 mg·kg⁻¹ and 21.90 mg·kg⁻¹, and those of male mice were calculated to be 58.62 mg·kg⁻¹ and 54.31 mg·kg⁻¹, respectively. The best fitting models for DNA damage induced by neodymium nitrate in female and male mouse hepatocytes were the exponential 5 model and the exponential 4 model, respectively, and the calculated BMD and BMDL were 27.15 mg·kg⁻¹ and 11.99 mg·kg⁻¹ for female mice, and 16.28 mg·kg⁻¹ and 10.47 mg·kg⁻¹ for male mice, respectively. The hill model was the best fitting model for DNA damage of gastric adenocytes in both female and male mice, and the calculated BMD and BMDL were 36.73 mg·kg⁻¹ and 19.92 mg·kg⁻¹ for female mice, and 24.74 mg·kg⁻¹ and 14.08 mg·kg⁻¹ for male mice, respectively. Conclusion Taken the micronucleus rate of bone marrow cells, DNA damage of liver cells and gastric gland cells as the end points of genotoxicity, the BMDL of neodymium nitrate is 10.47 mg·kg⁻¹, which can be used as the threshold of genotoxic effects induced by acute exposure to neodymium nitrate in mice.

Background The respiratory toxicity of inhaled polyhexamethylene guanidine (PHMG) has been extensively studied since the humidifier disinfectant incident. However, the impacts of inhalation of PHMG on the immune system are not comprehensively studied yet. Objective To explore the effects of inhalation of PHMG disinfectant aerosol on major immune organs and immune cells in mice. Methods Thirty male C57BL/6J mice (6-8 weeks old) were randomly divided into three groups: control, low-dose (0.1 mg·m⁻³ PHMG), and high-dose (1.0 mg·m⁻³ PHMG), with ten mice in each group. The mice were administered by oral-nasal inhalation of PHMG aerosol for 4 h per day, 5 d per week for 4 weeks consecutively. After designed treatment, venous blood was collected from the inner canthus of the eyes of mice and peripheral hematological indicators were measured with a blood analyzer. Then the mice were sacrificed by cervical dislocation and the lung, thymus, spleen, and femur were isolated. Lung, thymus, and spleen were weighed and organ coefficients were calculated, and single cell suspensions of thymus, spleen, and bone marrow were prepared to analyze lymphocytes phenotypes and proportions by flow cytometry. Results The body weight of mice in the high-dose group was lower than that of mice in the control group (P<0.01) from the 7th day of inhalation, and decreased by 15.74% compared with that of mice in the control group at the end of inhalation (P<0.01). The lung coefficients of both the low-dose and high-dose groups were higher than that of the control group (P<0.01), the thymus coefficient of mice in the high-dose group was lower than that of the control group (P<0.05), but the spleen coefficient did not change significantly (P>0.05). Leukocyte count [(1.49±0.22)×10⁹·L⁻¹], lymphocyte count [(0.96±0.36)×10⁹·L⁻¹] and its proportion [(63.13±14.96)%] in the peripheral blood of mice in the high-dose group were lower than those in the control group [(2.69±0.25)×10⁹·L⁻¹, (2.33±0.28)×10⁹·L⁻¹, and (86.23±3.40)%, respectively] (P<0.01), whereas red blood cell count [(12.32±0.46)×10¹²·L⁻¹], hemoglobin count [(175.25±4.65) g·L⁻¹], and hematocrit [(53.55±0.70)%] in the peripheral blood of mice in the high-dose group were higher than those in the control group [(11.11±0.37)×10¹²·L⁻¹, (160.67±4.04) g·L⁻¹, and (45.10±9.75)%, respectively] (P<0.05). Compared with the control group, the proportion of CD4+ CD8+ double-positive T cells decreased (P<0.05), the proportions of CD4+ T cells and CD8+ T cells increased (P<0.05), and the amounts of CD8+, CD4+ CD8+, CD4+, and CD4- CD8- cells decreased (P<0.05) in the thymus of mice of the high-dose group, the proportion of CD4+ T cells in the spleen of the high-dose group increased (P<0.05), the proportions and amounts of T cells, CD4+ T cells, and CD8+ T cells in the bone marrow of the high-dose group increased (P<0.05). Conclusion Inhalation of PHMG may cause thymic atrophy, disrupt T-lymphocyte development, and lead to an imbalance in the number of immune cells in the bone marrow, peripheral blood, and spleen, suggesting that inhalation of PHMG induces immune dysfunction.

Humans ; Receptors, Chimeric Antigen ; T-Lymphocytes ; Precursor Cell Lymphoblastic Leukemia-Lymphoma/therapy* ; Prognosis ; Immunotherapy, Adoptive ; Hematopoietic Stem Cell Transplantation ; Receptors, Antigen, T-Cell ; Recurrence