Objective To investigate the effect of insulin combined with Edaravone therapy on delayed cerebral vasospasm (CVS) after experimental subarachnoid hemorrhage (SAH).Methods 40 New Zealand white rabbits were randomly separated into sham-operated group (n=8),SAH group (n=10),insulin group (n=10) and insulin+Edaravone group(n=12).SAH models were made by injecting blood twice into cisterna magna.30 min later of the first blood injected,the insulin 0.2 U/kg was subcutaneous injected in insulin group and insulin+Edaravone group,3 times daily for 7 d;while,the insulin+Edaravone group received Edaravone 0.5 mg/kg (intravenous injected in ear vein) twice a day for 7 d.7 d after the model made,the basilar artery cross sectional area was measured and the pathological changes were obseved to estimate the degree of CVS.The expression of insulin receptor(InRa) in vascular endothelial cell was detected by immunohistochemical staining.Results The basilar artery cross sectional area between insulin+Edaravone group[(0.46?0.3)mm2]and the sham-operated group[(0.48?0.4)mm2] was no significant difference;but they were obviously bigger than those in SAH group[(0.25?0.3) mm2] and the insulin group [(0.30?0.3)mm2](all P

Objective:To isolate SD rat adipose-derived stem cells(ASCs)by suspended explant culture method.Methods: The healthy rat inguinal fat pads were obtained.The SD rat ASCs were isolated by suspended explant culture method and explant adherent culture method.The growth status and morphology were observed.The growth curve and cell surface markers CD29,CD44,CD45 of the 3rd passage cells were analyzed respectively by CCK-8,immunocytochemistry;the 3rd passage cells were induced individually by adipogenic differentiation medium and osteogenic differentiation medium,and cells were examined by oil red O staining and alizarin red staining.Results: The SD rat ASCs obtained by the two methods exhibited a spindle-shaped appearance and could rapidly expand.The cell growth curves were typical of S type.Immunocytochemistry analysis revealed that the third passage of SD rat ASCs were positive for CD29,CD44,but were negative for CD45.SD rat ASCs were positive for oil red O staining at 14 days after adipogenic induction,and positive for alizarin red staining at 14 days after osteogenic induction.Conclusion: Isolation of SD rat ASCs by suspended explant culture method is successfully established.The method is simple.It provides a new method for the isolation of SD rat ASCs in vitro.

Objective To study the effect of cryopreservation on some biological properties of rabbit adipose -de-rived mesenchymal stem cells (rADMSCs).Methods rADMSCs culture was isolated by tissue explants adherent meth-od.Morphology of the primary cells was observed by inverted microscopy .Immunophenotypes of the rADMSCs were deter-mined using flow cytometry .The third passage cells were preserved in liquid nitrogen for 6 months, and then were thawed , and subcultured to passage 7.The growth curves of the cryopreserved cells were analyzed by MTT assay , and the cryopre-served cells were cultured in adipogenic and osteogenic medium , with non-cryopreserved rADMSCs as a control group .The adipogenic and osteogenic abilities of the rADMSCs were evaluated by oil red O staining , alizarin red staining and alkaline phosphatase activity assay , respectively.Results The rADMSCs cultured in vitro exhibited a spindle-shaped appearance and rapid growth expansion .Flow cytometry analysis revealed that the third passage rADMSCs were CD 44-and CD90-posi-tive, but negative for hematopoietic cells surface marker CD 45.The growth curves of cells in the experimental and control groups were “S” shaped, showing a non-significant difference between the two groups (P>0.05).The oil red O staining and alizarin red staining results were positive at 2 weeks after adipogenic and osteogenic induction .The ALP activities of the two groups were increased with osteogenic induction time , with a non-significant difference (P>0.05).Conclusions Cryopreservation does not affect the growth and differentiation pluripotency of rADMSCs significantly .

significantly save the time of diagnosis and facilitate the clinical application of large samples.

Recording and extracting characteristic brain signals in freely moving animals is the basic and significant requirement in the study of brain-computer interface (BCI). To record animal's behaving and extract characteristic brain signals simultaneously could help understand the complex behavior of neural ensembles. Here, a system was established to record and analyse extracellular discharge in freely moving rats for the study of BCI. It comprised microelectrode and micro-driver assembly, analog front end (AFE), programmer system on chip (PSoC), wireless communication and the LabVIEW used as the platform for the graphic user interface.
Animals ; Behavior, Animal ; physiology ; Brain ; physiology ; Electroencephalography ; methods ; Microelectrodes ; Rats ; Signal Processing, Computer-Assisted ; instrumentation ; Telemetry ; instrumentation ; User-Computer Interface

OBJECTIVETo compare the efficacy and safety between laparoscopic total extraperitoneal prosthetic (TEP) and Lichtenstein herniorrhaphy in the treatment of inguinal hernia (IH).

METHODSClinical data of 158 IH patients at our hospital from October 2015 to October 2016 were retrospectively analyzed. Among 158 patients, 85 patients underwent TEP (TEP group), and 73 underwent Lichtenstein herniorrhaphy(Lichtenstein group). Patients of TEP group received general anesthesia. The space between suprapubic space and iliac fossa was separated with attention to protect the spermatic cord during the operation so as to avoid excessive separation. When the patch was placed, the internal margin exceeded the contralateral pubic tubercle, the lateral margin reached the anterior superior iliac spine, there was at least 2 cm overlap between the superior margin and the joint tendon, the lower margin was inserted into the suprapubic bladder space(Retzius space), and the lateral lower margin was at least 6 cm to internal ring. Lichtenstein group received local anesthesia or continuous epidural anesthesia and all underwent routine Lichtenstein herniorrhaphy. Operative time, postoperative conditions and recurrence were compared between two groups.

RESULTSAmong these 158 patients, 129 were male and 29 were female, with an age ranging from 26 to 75 years (median 42 years). Baseline data were not significantly different between two groups(all P>0.05). There was no perioperative death and serious complications in two groups. The difference was not statistically significant in operative time[(47.6±5.4) minutes vs. (48.9±6.3) minutes, t=0.238, P=1.024]. But TEP group was better than Lichtenstein group in terms of intraoperative blood loss [(7.53±2.31) ml vs. (11.41±4.49) ml, t=5.783, P=0.032], postoperative bedtime [(1.25±0.32) days vs. (2.83±0.52) days, t=5.294, P=0.041], postoperative hospital stay [(2.38±1.14) days vs. (3.46±1.81) days, t=5.482, P=0.037], and postoperative pain score (1.27±0.47 vs. 3.42±1.93, t=5.639, P=0.034). Follow-up was 12-16 months (median 10 months). In TEP and Lichtenstein group after operation, incision infection occurred in both 1 case, edema of the scrotum or labia minora developed in 2 patients and 3 cases respectively, chronic pain was found in one and 2 cases respectively. The patients were followed up for 5 to 16 months(median 10 months), and recurrences was observed in 1 case both in TEP group and Lichtenstein group respectively.

CONCLUSIONLaparoscopic total extraperitoneal prosthetic is safe and effective in the treatment of inguinal hernia, which is better than Lichtenstein herniorrhaphy in fast recovery, less trauma and more comfort, and worthy of clinical promotion.

The nervous system is one of the most complicated organ systems in invertebrates and vertebrates. Down syndrome cell adhesion molecule (DSCAM) of the immunoglobulin (Ig) superfamily is expressed widely in the nervous system during embryonic development. Previous studies in Drosophila suggest that Dscam plays important roles in neural development including axon branching, dendritic tiling and cell spacing. However, the function of the mammalian DSCAM gene in the formation of the nervous system remains unclear. Here, we show that Dscam ( del17 ) mutant mice exhibit severe hydrocephalus, decreased motor function and impaired motor learning ability. Our data indicate that the mammalian DSCAM gene is critical for the formation of the central nervous system.
Animals ; Cell Adhesion Molecules ; genetics ; metabolism ; Corpus Callosum ; metabolism ; pathology ; Genotype ; Hydrocephalus ; genetics ; metabolism ; pathology ; Mice ; Mice, Knockout ; Motor Activity ; genetics ; physiology ; Mutation

Background and purpose:As one of the most fatal malignant tumors in China, the morbidity and mortality of lung cancer remain high. Early diagnosis and normative treatment is the key to improve the prognosis of lung cancer. The aim of this study was to explore the practice of early lung cancer screening with low-dose spiral computed tomography (CT) based on the current situation in community health service, with integration of superior resources of med-ical institutions at all levels in Shanghai.Methods:From Aug. 2013 to Aug. 2014, we screened high-risk population in selected communities of Minhang District, Shanghai, for early diagnosis of lung cancer with low-dose spiral CT combined with multidisciplinary comprehensive treatment models including minimally invasive surgery, exploring the medical service network covering prevention, diagnosis, treatment, rehabilitation and follow-up.Results:Screening population is 11 332 (male 7 144, female 4 188); Twenty-nine cases with pathological diagnosis of malignant tumor, including 27 cases of pri-mary lung cancer, 1 case of lung metastasis, 1 case of breast cancer. The morbidity of primary lung cancer is 238.26×10-5. There were 22 cases of Stage 0-Ⅰ lung cancer accounting for 81.48% of all diagnosed primary lung cancer.Conclusion:Based on community health service, screening with low-dose spiral CT could improve the early diagnosis rate of lung can-cer with feasibility and validity, which could be applicable in qualified eligible medical center and community in China.

Objective:To explore the airway pathogen characteristics and examine the correlation between donor-derived pathogens and post-transplant outcomes in patients after lung transplantation (LT).Methods:Between January 1, 2015 and December 31, 2019, retrospective review was conducted for clinical and microbiological data of 88 LT recipients.Airway pathogen percentage of different microorganisms and evolution of drug-resistance were examined.Drug-resistant pathogen positive group (n=71) and negative group (n=17) were assigned according to whether or not drug-resistant pathogens were detected.Survival analysis was conducted by Log-rank with 3-year follow-ups.Between April 11, 2020 and September 5, 2020, prospective study was conducted in 14LT recipients.The potential pathogenic bacteria from donor lungs were detected by metagenomic next generation sequencing and the impact of those bacteria was examined on 1-year post-transplantation outcome in 2020.Microbial diversity and richness were shown with Shannon index.The outcome variables included heart rate, neutrophil count, lymphocyte count, immunoglobulin level and pulmonary spirometry.ANOVA and Pearson's correlation analysis were performed for elucidating the relationship between airway microbiota and post-LT outcomes.Results:From 2015 to 2019, 88 recipients were recruited and 992 strains of airway pathogens were isolated, including bacteria 796 strains and fungi 196 strains.Gram-negative bacteria (704 strains) accounted for 88.4% of all bacteria.The detection rates of Gram-positive bacteria, Klebsiella pneumonia (Kp), Acinetobacter baumannii (Ab), Stenotrophomonas maltophilia and Candida increased in 2019 than that in 2015 (8.2% vs. 5.3%, 13.6% vs. 13.2%, 33.2% vs. 17.5%, 6.5% vs. 5.3%, 26.6% vs. 20.2%). Drug resistance rate of Kp to imipenem was 68.18% in 2019 and drug resistance rate of Ab to imipenem 98.44%.The 3-year survival rate was 46.3% and 35.3% in drug-resistance positive and negative groups and the difference was insignificant ( P=0.410). Fourteen recipients were enrolled in 2020.Potential pathogenic bacteria could be detected in all donor samples.Five recipients carried the same bacteria and two died during 1-year follow-up.Nine recipients did not carry the donor-derived pathogens and two died during 1-year follow-up.The diversity of donor/recipient-derived airway microbiota (Shannon index) showed no correlation with the outcomes of 1-year follow-up by Pearson's correlation test. Conclusions:Gram-negative bacteria predominated in airway pathogens of recipients post-LT.The drug resistance rate to imipenem remained high.The donor/recipient-derived pathogen isolates showed no correlation with immediate outcomes post-LT.

ObjectiveTo investigate the promotional effect of astragaloside on the repair and healing of chronic non-healing wounds and its mechanism. MethodA total of 60 male SD rats were constructed with full-layer skin defect wounds on the back， and except for the control （Con） group， the rest were constructed with non-healing wounds， which were then randomly divided into the sham-operation （sham） group， the low-dose astragaloside group， the high-dose astragaloside group， the astragaloside + LY294002 ［phosphatidylinositol 3-kinase （PI3K） inhibitor］ group， and the astragaloside + EX527 ［silencing regulatory protein 1 （SIRT1） inhibitor］ group. The percentage of wound area in each group was observed on the 2nd， 4th， 6th， and 8th days after wound molding. Collagen type Ⅰ alpha 1 （COL1A1） and alpha smooth muscle actin （α-SMA） expressions in the wound tissue were detected by immunofluorescence. Hematoxylin and eosin （HE） staining was performed to determine the pathological structure of the wound. The mRNA expression of inflammatory factors in the wound was measured by real-time polymerase chain reaction （Real-time PCR）， and the expression of proteins related to the SIRT1/ nuclear factor （NF）-κB and PI3K/protein kinase B （Akt） signaling pathways in the wound was tested by Western blot. ResultCompared with the sham group， the percentage of postoperative wound area of rats in both low-dose and high-dose astragaloside groups gradually decreased with time， and the efficacy of the high-dose astragaloside group was better. Compared with the Con group， the fluorescence intensity of COL1A1 in wound tissue of the sham group decreased， while the expression of α-SMA increased. The epithelial tissue was severely damaged， with an increase in the thickness， and a large number of inflammatory cells were seen in the infiltration. The mRNA expression of tumor necrosis factor （TNF）-α， interleukin （IL）-1β， IL-6， and inducible nitric oxide synthase （iNOS） was elevated. The protein expression of NF-κB p65， p-PI3K/PI3K， and p-Akt/Akt was elevated， while SIRT1 expression was decreased （P<0.05）. Compared with the sham group， the fluorescence intensity of COL1A1 and α-SMA increased after astragaloside treatment. The number of epithelial cells increased， and the thickness decreased. The inflammatory cells decreased， and the amount of collagen increased. The mRNA expression of TNF-α， IL-1β， IL-6， and iNOS was decreased， and the protein expression of NF-κB p65， p-PI3K/PI3K， and p-Akt/Akt was decreased. SIRT1 was elevated， and the effect was better in the high-dose astragaloside group （P<0.05）. Compared with the high-dose astragaloside group， inhibition of the PI3K/Akt and SIRT1 pathways by LY294002 and EX527 prevented the therapeutic efficacy of astragaloside on chronic non-healing wounds. ConclusionThe topical application of astragaloside significantly promotes the healing of chronic non-healing wounds in rats， and the mechanism may be related to the activation of the PI3K/Akt pathway and the SIRT1/NF-κB pathway.