Objective:To observe the effect of old RBCs transfusion on cognitive function in rats and the improvement effect of mi -nocycline.Methods: Male SD rats at the age of 6 months were randomly divided into 4 groups.The RBCs were obtained from male rats by centrifuging the total blood and stored at 4℃.The rats of fresh RBCs group (group F) were transfused with the RBCs stored for 1 day.The rats of old RBCs group (group O) were transfused with the RBCs stored for 7 days.The rats of treatment group (group T) received 40 mg· kg-1 minocycline with intraperitoneal injection before the transfusion .The rats of the control group ( group C) were transfused with the normal saline .The brain levels of IL-1βand IL-6 were determined with Quantikine ELISA kits in 24 hours after the blood transfusion (n=6).The rats were subjected to Barnes maze tests after 1 week of the blood transfusion (n=10).Results:The brain levels of IL-1βand IL-6 in group O were higher than those in group C and F (P<0.05), which were lower in group T than those in group O(P<0.05).The rats of group O spent longer time finding the target box than those of group C and F in the Barnes maze (P<0.05), and the time was shorter in group T than that in group O (P<0.05).Conclusion: Old RBCs transfusion plays a role in neuro-inflammation and induces cognitive dysfunction in rats , which may be improved by minocycline .

BACKGROUND: The transplantation of allogeneic cartilage has local immunological rejection, and it is necessary to further reduce the rejection to promote its application in clinic, thus it is significant to perform a series of experiments to induce local immune privilege.OBJECTIVE: To observe the in vivo growth of tissue engineered allogeneic cartilage reconstructed by chondrocytes transfected with recombinant retroviral vector pLNCX2-FasL.DESIGN: A randomized controlled observation.SETTING: Shanghai Jiao Tong University.MATERIALS: Thirty-six allogeneic New Zealand rabbits as recipients and 45 1-week-old chinchillas as donors, either sex,were purchased by the experimental animal center of Chinese Academy of Sciences. Amphotropic recombinant retrovirus coated cell line PT67 was purchased from Clontech Company; Dulbecco's modified eagle medium (DMEM), fetal bovine serum (FBS), G418 and Polybrene were bought from GIBCO BRL.METHODS: The experiment was carried out in original Shanghai Second Medical University from January 2000 to July 2005. The New Zealand rabbits were randomly divided into three groups: FasL-transfected group (n =12), untransfected group (n =12) and blank control group (n =12). The rabbit allogeneic cartilages were constructed by the compound of pLNCX2-FasL transfected chondrocytes and tissue engineered material of pluronic F-127. ① Gross observation and mass changes of the grafts: Corresponding materials were infused subcutaneously, the grafts were removed at 1, 2 and 3 months after transplantation for gross observation and the mass changes. ② Staining observation: The grafts were removed at 1, 2 and 3 months after transplantation, then prepared into sections, and observed by hematoxylin and eosin (HE), Safranin'O and Masson's trichrome stainings. ③ Antibody detection: Blood samples (1 mL) were collected at 1 and 2 months after transplantation, the chondrocytes of the chinchillas were lysed freezingly with lysis antigen as the mixed antigen, and separated by electrophoresis in agarose medium, then acted with serum of recipient to observe whether corresponding antibody generated. ④ Complement dependent cytotoxicity (CDC) test: The chondrocytes of chinchillas were prepared into cell suspension (2×109/L), and then seeded into 96-well plate, attached grew for 24 hours, then recipient serum was added for the CDC test, and the percentage of apoptotic cells was counted under microscope.MAIN OUTCOME MEASURES: ① Gross observation and mass changes of the grafts;② Histological changes; ③ Results of the antibody detection; ④ Percentage of apoptotic cells.RESULTS: All the 81 rabbits were involved in the analysis of results. ① Gross observation and mass changes of the grafts: Two weeks after inoculation, there were obvious nod formations at the inoculated sites, but no nod formed in the blank control group. The new cartilage tissues became smaller gradually and completely disappeared at 4 months in the untransfected group, whereas those in the FasL-transfected group became smaller, but still existed after 7 months. The masses of grafts in the FasL-transfected group were higher than those in the untransfected group (P ＜ 0.05). ②Histological observation: Plenty of lymphocytic infiltrations around cartilage tissue could be observed in the untransfected group, and obviously decreased in the FasL-transfected group. No lymphocyte was observed inside the chondrocytes.Masson's trichrome staining was performed, and it was observed under light microscope that the small white parts in the middle were immature chondrocytes, and there were green collagen around most of the mature chondrocytes. Safranin O staining showed strong positive reaction, suggested that there were rich glycosaminoglycan in matrix. ③ Antibody detection: The chondrocytes of the chinchillas were lysed freezingly with lysis antigen as the mixed antigen, then acted with serum of recipient, and the results showed that no corresponding antibody generated. ④ Percentage of apoptotic cells: The percentages of serum CDC apoptotic cells in the FasL can ransfected group, untransfected group and blank control group were 5%, 6% and 1%, which were all negative.CONCLUSION: Rabbit allogeneic chondrocytes transfected with recombinant retroviral vector pLNCX2-FasL can reconstruct tissue engineered cartilage, and can postpone the degeneration by 3 months.

Objective To establish the gas chromatographic(GC) fingerprint of oleic acid of Whitmania pigra Whitman for its quality control. Methods Ten batches of Whitmania pigra from different sources and processed by different methods were analyzed with Agilent 6890N gas chromatography detector on DB-WAX(30 mm × 0.32 mm × 0.25μm)column at the vaporizing temperature of 270℃, column temperature of 130℃and flame ionization detector (FID) temperature of 280℃. We used a software of Similarity Evaluation System for Chromatographic Fingerprint of TCM(Version of 2004A) published by Chinese Pharmacopoeia Commission to calculate GC similarity. Results Oleic acid content of Whitmania pigra processed by different methods had significant differences (F2,7 = 7.350, P = 0.019). The oleic acid content of samples dried after washing with clean water significantly differed from that of the samples processed by alumen or the slices dried naturally(P = 0.021, P= 0.009). The similarity of the fingerprints was in the range of 0.458 - 0.998. The similarity of samples from Lipu of Guangxi Province was the lowest. Conclusion The fingerprints of most of the samples have very high similarity. The established GC fingerprints can be used to effectively identify the qualified or inferior Whitmania pigra products, which will provide some reference for the quality control of Whitmania pigra.

High extracellular potassium can induce spreading depression-like depolarizations, elevations of extracellular glutamate and even neuronal death in normal brain. To investigate the contribution of high potassium in vivo, a microelectrode arrays ( MEAs ) probe integrated with recording sites for glutamate concentration (50í150 μm) and local field potential ( LFP) ( diameter=15 μm) was fabricated by Micro-electro-mechanical-systems ( MEMS) technologies. We implanted the MEA probe acutely in the rat brain and exposed the brain to a high potassium solution. During these multi-modal recordings, it was observed that high potassium elevated extracellular glutamate while suppressing the LFP irreversibly. This is one of the first studies in which a dual mode MEA probes is applied in vivo for neuronal death, and it is concluded that our MEA probes are capable of examining specific spatiotemporal relationships between electrical and chemical signaling in the brain.

ObjectiveTo investigate the impacts of neural stem cells (NSCs) transplantation on spinal pathology and ultrastructure after spinal cord injury (SCI) in rats and probe into the protective role of tacrolimus (FK506) on neural regeneration.MethodsCompressive SCI at T8 was induced in the adult SD rats,which were randomly assigned to the control group,FK506 group,NSCs group and NSCs + FK506 group.The differences of neural regeneration in each group were compared at days 7,14,28 and 56 after injury by motor evoked potentials ( MEP),HE staining,immunohistochemical staining,ultrastructure observation and image analysis of the myelinated fiber. ResultsThe MEP latency in the NSCs + FK506 group was significantly shorter than that in other groups at day 28 ( P ＜ 0.05 ).HE staining revealed that only local necrosis presented in the NSCs + FK506 group at day 56.More BrdU and NF-200 positive cells were detected with immunohistochemical staining in the other three groups as compared with the control group.Moreover,the positive cells in the NSCs + FK506 group also outnumbered the FK506 group and NSCs group.Electron microscope scan showed edema under the membrane of large myelin sheath in the control group,and classic new myelin sheath and neuraxis in the NSCs + FK506 group at day 56.The regeneration of the nerve fiber in the NSCs + FK506 group was better than that of other three groups (P ＜0.01 ).ConclusionAfter NSCs transplantation for SCI rats,the early combination use of FK506 can improve the pathology and ultrastructure of the regenerative nerve fiber and is conducive to nerve regeneration.

Objective We investigated if the proliferative capacity of endothelial progenitor cells was affected by hypoxia and exercise. Methods Twenty four male Sprague-Dawley rats were randomly and averagely divided into 4 groups: (1) living at low altitude (LL), (2) living and training at low altitude (LLTL), (3) living at high altitude (LH), and (4) living at high altitude and training at low altitude (LHTL). Eight-week incremental treadmill exercise and hypoxic simulation were used to establish LHTL animal model. Mononuclear cells from peripheral blood were obtained by density gradient centrifugation 12 hours by the end of 8-week experiment. The cells were suspended in conditioned medium 199 for culturing in vitro. Their phenotypes were confirmed by uptake of acetylated LDL and binding of fluoresce in isothiocyanate (FITC)-labeled Ulex europaeus agglutinin 1 (UEA-1) lectin. Inverted microscopic observation was used to identify the morphological changes in endothelial progenitor cells and measure the cell count. Results Adherent cells and early CFUs in groups LLTL, LH and LHTL increased more obviously than in group LL(P<0.01), whereas the number of attached cells between group LH and LHTL was not different (P>0.05). Conclusion Proliferative capacity of endothelial progenitor cells can be promoted by both hypoxic stimulation and exercise, and the promotion is more significant if combination of hypoxia and exercise was employed simultaneously.

Objective To observe the relationship between the integrity of Willis circle and aneurysm with MSCT angiography.Methods CTA data of 580 patients with intracranial vascular lesions were retrospectively analyzed.The morphological structure of Willis circle and the occurrence of aneurysm were observed based on axial thin-layer,VR,MIP and MPR images.According to the integrity,Willis circle were divided into type Ⅰ (Willis circle complete),type Ⅱ (the anterior circulation complete but the posterior circulation incomplete),type Ⅲ (the anterior circulation incomplete but the posterior circulation complete) and type Ⅳ (both the anterior and posterior circulation incomplete) for statistical analysis.Results For the classification of Willis circle,there were 118 cases of type Ⅰ (118/580,20.34％),344 of type Ⅱ (344/580,59.31％),25 of type Ⅲ (25/580,4.31％) and 93 cases of type Ⅳ (93/580,16.03％).The incidence of Ⅰ-Ⅳtypes of Willis circle with aneurysm accounted for 16.10％ (19/118),14.83％ (51/344),32.00％ (8/25) and 23.66％ (22/93),respectively.The incidences of aneurysm in patients with different types of Willis circlewere statistically significant (x2=8.013,P=0.046).There was statistical difference of the type of Willis circle between different genders (x2=12.631,P=0.006),and the incidence of aneurysm in females was higher than that in males (25.00％ [63/252] vs 11.28％ [37/328];x2 =18.80,P＜0.025).Conclusion Most Willis circle were not complete,and incomplete Willis circle aneurysm tended to occur in females with higher aneurysm rate.

Objective To investigate the feasibility of using anti-sense RNA against classⅡmajor histocompatibility complex(MHCⅡ)transactivator(CⅡTA),which might regulate MHCⅡexpression,to suppress the relative immune response.Methods Stable transfectants of dermal fibroblasts with pDarⅡ(pDarⅡ-D)were tested for the expression of classic MHCⅡ(HLA-DR,-DP,-DQ)antigens induced with recombinant human interferon-gamma(IFN-?).mRNA abundance of CⅡTA,and classic MHCⅡwas mea-sured by RT-PCR.IL-2mRNA expressed in T cells,stimulated by transfected dermal fibroblasts,was de-termined by mixed lymphocyte reaction.Results When induced with IFN-?,the expression of HLA-DR and-DP antigens on pDarⅡ-D was reduced by95.63%and87.89%,respectively.Meanwhile,the mRNA contents of CⅡTA and classic MHCⅡwere decreased significantly(P

Objective To investigate the application value of DynaMesh-IPST stoma dedicated mesh in parastomal hernia repair.Methods The retrospective cohort study was conducted.The clinical data of 281 patients with parastomal hernia of abdominal wall who were admitted to Beijing Chao-Yang Hospital of Capital Medical University between March 2013 and April 2017 were collected.Of 281 patients undergoing laparoscopic combined with open parastomal hernia repair with artificial materials,151 using DynaMesh-IPST stoma dedicated mesh and 130 using ordinary anti-adhesive mesh were respectively allocated into the DynaMesh-lPST mesh group and ordinary mesh group.Observation indicators:(1) intra-and post-operative situations;(2) follow-up.Follow-up using outpatient examination and telephone interview was performed to detect hernia recurrence and long-term complications at 1-,3-,6-month and 1 year postoperatively up to June 2018.Measurement data with normal distribution were represented as (x)±s and comparison between groups was done by the t test.Measurement data with skewed distribution were described as M (P25,P75) and M (range),and comparison between groups was done using rank sum test.Comparison of count data between groups was analyzed using the chi-square test.Results (1) Intra-and post-operative situations:281 patients underwent successfully laparoscopic combined with open parastomal hernia repair with artificial materials.The operation time,volume of intraoperative blood loss and duration of postoperative hospital stay were 100 minutes (60 minutes,120 minutes),(34± 15)mL,17 days (13 days,24 days) in the DynaMesh-IPST mesh group and 100 minutes (85 minutes,120 minutes),(42± 36)mL and 20 days (16 days,25 days) in the ordinary mesh group,with statistically significant differences between groups (Z=2.166,t=2.654,Z=2.795,P＜0.05).The postoperative incisional infection and intestinal leakage were respectively detected in 18 and 2 patients in the DynaMesh-IPST mesh group and 36 and 7 patients in the ordinary mesh group,showing a statistically significant difference of incisional infection between groups (x2 =11.194,P＜0.05),and no statistically significant difference of intestinal leakage between groups (x2 =4.234,P＞ 0.05).Of 54 patients with postoperative incisional infection,4 were cured after removing mesh and clearing up infection,50 were cured after dressing change,local debridement and drainage.(2) Follow-up:279 of 281 patients including 149 in the DynaMesh-IPST mesh group and 130 in the ordinary mesh group were followed up for 20-44 months with a median time of 32 months.During the follow-up,10 patients had hernia recurrence including 4 in the DynaMesh-IPST mesh group and 6 in the ordinary mesh group.Six of them underwent reoperation (4 with keyhole hernia recurrence,1 with mesh bulging out because of abdominal weakness at stoma,1 with parastomal recurrence after mesh removal due to intestinal leakage) and 4 had regular reexamination after fixation with abdominal belt.There was no statistically significant difference of hernia recurrence between groups (x2 =0.318,P＞0.05).Twenty-seven patients (14 in the DynaMesh-IPST mesh group and 13 in the ordinary mesh group) with intestinal obstruction were improved by conservative treatment,showing no statistically significant difference between groups (x2 =0.043,P＞0.05).Ten patients (6 in the DynaMesh-IPST mesh group and 4 in the ordinary mesh group) with stoma stenosis were improved after local stoma remodeling,showing no statistically significant difference between groups (x2=0.007,P＞ 0.05).Ninteen patients (4 in the DynaMesh-IPST mesh group including 1 complicated with hernia recurrence and 15 in the ordinary mesh group including 2 complicated with hernia recurrence) with stoma prolapse were improved after local stoma remodeling,showing a statistically significant difference between groups (x2 =8.756,P ＜ 0.05).Conclusion Parastomal hernia repair with DynaMesh-IPST stoma dedicated mesh is safe and feasible,with satisfactory effects.

Objective:To summarize the characteristics of autosomal recessive axonal neuropathy with neuromyotonia (ARAN-NM) caused by HINT1 gene mutation. Methods:Retrospective case summary.Clinical data of 2 Tibetan siblings diagnosed with ARAN-NM in the Department of Pediatrics of Peking University First Hospital in August 2023 were retrospectively analyzed.A review of literature reporting relevant Chinese patients was conducted.Results:The proband and her elder brother were aged 13 and 19, respectively.Both developed abnormal gait at the age of 11, followed by varus, claudication, and weak thumb strength.The proband also had neuromyotonia.Physical examinations showed that the proband and her elder brother had decreased muscle strength of the extremities, mainly in the thumbs and distal ends of lower limbs.The distal muscles of the proband′s lower extremities and the muscles of both hands of the proband′s elder brother were atrophied.Both feet showed talipes equinovarus in the proband and her elder brother.The proband′s electromyography (EMG) showed peripheral nerve injuries (motor and sensory axonal involvement, especially in distal ends) and myotonic potentials.The trio-whole exon sequencing detected homozygous pathogenic variation in HINT1 gene in both the proband and her elder brother, who were diagnosed as ARAN-NM based on c. 169A>G (p.K57E). After the Carbamazepine treatment, the proband′s neuromyotonia, numbness and weakness were relieved.Both the proband and her elder brother underwent orthopaedic surgery and rehabilitation.Their foot deformities and gait were significantly improved.Two Chinese literatures (2 patients) and four English literatures (8 patients) were retrieved.Including the proband and her elder brother in this study, there were 12 ARAN-NM patients, 10 of whom had clinical data.The ages of onset and diagnosis were 2-16 (1 case unknown) and 13-33 years old, respectively.Myasthenia was present in 9 patients, especially in distal ends.Eight patients were complicated with neuromyotonia, nine patients with muscle atrophy, seven patients with foot deformity, and two patients with sensory disturbance.Creatine kinase(CK) was elevated in all 9 patients tested or CK.EMG showed neurogenic injuries in all patients and neuromyotonia discharge in six patients.Three patients were treated with Carbamazepine, and some symptoms were relieved.Missense/nonsense mutations were found in the 12 patients, and the high-frequency variation was c. 112T>C (p.C38R). Conclusions:ARAN-NM is a rare autosomal recessive neuromuscular disease caused by HINT1 gene mutation.There is no ethnic difference in clinical manifestations, mainly distal limb weakness with neuromyotonia.Carbamazepine can alleviate some symptoms, and orthopaedic surgery can improve foot deformity and gait.