Objective To establish a three- dimensional hindlimb gait data toolkit (THGT) for healthy and spinal cord injured (SCI) non-human primate (rhesus monkey) based on Matlab to realize upload of original data, automatic gait division, calculation and drawing of multiple gait parameters, etc. Methods Vicon system was used to collect three-dimensional hindlimb gait data of healthy and SCI (after 6 weeks) rhesus monkey to obtain the kinematics data of both hindlimbs in continuous strides. It was analyzed with THGT to process the gait division, calculation and drawing of multiple gait parameters. Results THGT read the data, distinguished cycles of gait, calculated 140 kinds of gait parameters and drew graphs of the results. Conclusion THGT extends the universality of the Vicon data, realizes automatically gait division and friendly interactive interface, and puts out the visible results.

Objective To find methods to isolate and purify plasminogen activator (Pla) from artificial culture of Yersinia pestis. Methods Ultrasonication and urea extracting combined by ammonium sulfate salting-out were tried to extract Pla. High performance liquid chromatography(HPLC) was used to purify Pla. The first step was ion exchange and the second was gel filtration, Preparative electrophoresis was used to purify Pla, too. The enzyme activity of the isolated or purificated Pla was detected. Results Both 50% - 60% saturated ammonium sulfate deposition of supernatant of plague bacilli ultrasonication and 0 - 10% saturated ammonium sulfate deposition of supernatant of plague bacilli powder soaked by urea had three bands(Mr about 31×103, 35×103 and 37×103) and lysis rings were 6.5 and 7.2 mm in diameter respectively when the enzyme activity was detected. Pla purified by HPLC was mainly composed of three bands(Mr about 31×103, 35×103 and 37×103), occupying more than 80% of total protein weight and lysis ring was 5.0 mm in diameter. Pla purified by preparative electrophoresis mainly consisted of three bands(Mr about 31×103, 35×103 and 37×103) with other proteins of low concentration nearby, no lysis ring was detected. Conclusions Pla is collected by the methods of ultrasonication and urea extracting. Priliminary purification of Pla can be achieved by HPLC and preparative electrophoresis.

Objective To evaluate the association of islet autoantibodies [ glutamic acid decarboxylase antibody(GADA),protein tyrosine phosphatase antibody(IA-2A)and insulin autoantibodies(IAA)1 with HLA- DQ genotypes in the first-degree relatives of autoimmune type 1 diabetes mellitus.Methods This was a cross- sectional and case-control study.Three hundred and fifty-one first-degree relatives with normal glucose tolerance of patients with type 1 diabetes mellitus and 376 healthy controls were recruited and measured for GADA,IA-2A and IAA by radioligand assay,and 156 first-degree relatives of patients with autoimmune type 1 diabetes mellitus and 278 controls were typed for genetic polymorphisms of HLA-DQ with PCR sequencing-based typing method.Results (1)DQA1*03,DQBI*0303,*0401 alleles and DQA1 * 03-DQBI * 0303,DQA1 * 05-DQBI * 0201,DQA1 * 03-DQBI * 0401 haplotypes were significantly increased in the first-degree relatives of autoimmune type 1 diabetes mellitus(P

Objective Diminished ovarian reserve (DOR) severely affects the life of women and the estrogen replacement therapy for it has obvious adverse effects. This article aimed to study the effect of polygoni multiflori radix preparata (PMRP) on DOR in rats and provide a therapeutic option for clinical medication. Methods Sixty female SD rats were randomly divided into six groups of equal number,normal control,DOR model control,high-dose PMRP (4 g/kg),medium-dose PMRP (2 g/kg),low-dose PMRP (1 g/kg),and positive control. The DOR model was established by gavage of tripterygium glycosides as 75 mg/kg every morning,followed by administration of PMRP in the PMRP groups,Estradiol valerate at 0.18 mg/kg in the positive control group and distilled water in the model control group in the afternoon,all for 30 consecutive days. The estrous cycle of the rats was observed,the levels of serum estradiol (E2),follicle-stimulating hormone (FSH),luteinizing hor-mone (LH),anti-Müllerian hormone (AMH) and inhibin-B (INH- B) were determined by ELISA,the ovarian and uterine indexes were obtained,and the ovarian morphology was observed by HE stai-ning,and the counts of follicles at different stages were recorded. Results Compared with the normal controls,the DOR model rats showed modeling time-related lengthening,irregularity and even disorder of the estrous cycle,with a few epithelial cells or keratino-cytes and leucocytes on the vaginal smear at 11-30 days. The estrous cycle was normal in the PMRP and positive control groups at 1-10 days and relatively prolonged at 11-30 days. In comparison with the normal control group,the DOR model rats exhibited a signifi-cantly decreased levels of serum E2 ([302.6±42.9] vs [155.7±46.8] pg/mL,P<0.05) and INH-B ([494.5±84.1] vs [299.2± 106.8] pg/mL,P<0.05) but increased levels of FSH ([7.2±0.5] vs [21.7±1.2] mIU/mL,P<0.05) and LH ([17.4±1.2] vs [25.0±1.0] mU/mL,P<0.05). The INH-B level was markedly elevated in the PMRP and positive control groups as compared with that in the DOR models (P<0.05). The counts of follicles and corpora lutea were remarkably lower in the DOR model rats (P<0.05) while that of developing follicles markedly higher in the PMRP and positive control groups than in the normal control group (P<0.05). The numbers of atretic follicles+corpora lutea were significantly increased in the high-dose PMRP group but decreased in the low-dose PMRP group (P<0.05) and positive controls (P<0.05). The counts of primordial and developing follicles were dramatically higher in the PMRP and positive control groups than in the DOR model controls (P<0.01) and so were the numbers of atretic follicles+corpora lutea in the high-and medium-dose PMRP groups (P<0.05). Conclusion Polygoni multiflori radix preparata can effectively protect the reproductive function of female rats by inhibiting tripterygium glycosides-induced toxicity to the ovary.

Objective To observe the influence of spinal cord injury on lower limbs weight support capacity,and the changes with time. Methods Six adult female rhesus monkeys with thoracic(T7-9)right spinal cord hemi-section were measured the plantar pressure ratio of both lower limbs with Foot-Scan system before,and six and twelve weeks after operation. Results There was no statistical difference between both sides of limbs before operation(Z=-1.330,P>0.05),while the plantar pressure ratio was more on the left limbs six and twelve weeks after operation(Z>4.783,P<0.001).The plantar pressure ratio of right lower limb became less and less during observation(Z=3.191,P<0.001). Conclusion The weight support capacity of affected limbs is injured after spinal cord hemi-section in monkeys, and would become worse without intervention.

OBJECTIVETo construct differential expression profiles of adenoid cystic carcinoma cell lines for screening candidate genes related to metastasis and to verify some candidate genes in adenoid cystic carcinoma.

METHODSRestriction fragments differential display PCR (RFDD-PCR) was used to set up gene expression profiles of adenoid cystic carcinoma cell lines-ACC-M and ACC-2, with high and low metastasis potential respectively. Candidate genes were screened through bioinformatics analysis. Then, a gene family of these candidate genes was checked using semi-quantitative reverse transcription-PCR(RT-PCR).

RESULTSTwo gene expression profiles including 5420 gene fragments were constructed, 12 genes of a family called matrix metalloproteinase genes (MMPs) were observed obvious differentially expressed between two cell lines. Results of semi-quantitative RT-PCR also identified this different expression of MMP2,MMP7,MMP9,MMP14,MMP15 and MMP24.

CONCLUSIONThe construction of gene expression profiles of ACC-M and ACC-2 cell lines makes the foundation for seeking the target genes of adenoid cystic carcinoma. MMP2,MMP7,MMP9 and MMP15 may be relevant with carcinogenesis, development and metastasis of adenoid cystic carcinoma, and different metastasis potential may result from different subtype of MMPs gene family.

Carcinoma, Adenoid Cystic ; enzymology ; genetics ; pathology ; Cell Line, Tumor ; Gene Expression Profiling ; Gene Expression Regulation, Enzymologic ; Gene Expression Regulation, Neoplastic ; Humans ; Matrix Metalloproteinase 15 ; genetics ; Matrix Metalloproteinase 2 ; genetics ; Matrix Metalloproteinase 7 ; genetics ; Matrix Metalloproteinase 9 ; genetics ; Matrix Metalloproteinases ; genetics ; Neoplasm Metastasis ; Reverse Transcriptase Polymerase Chain Reaction ; methods

OBJECTIVETo evaluate the clinical result of arthroscopic anterior cruciate ligament (ACL) reconstruction with either allo- or auto- bone-patellar tendon-bone (B-PT-B) grafts.

METHODSFrom February 2002 to January 2006, 142 of 187 cases of ACL ruptures who received ACL reconstruction with B-PT-B grafts were studied retrospectively. There were 93 male and 49 female whose age was from 15 to 57 years (mean 26 years). Patients were divided into 2 groups by graft selection: 38 patients with autograft and 104 with allograft. Clinical results were evaluated according to IKDC, Lysholm, Irgang and Larson scales. Sub-items of scales such as pain, swelling and laxity were specifically evaluated.

RESULTSAll of the patients were followed up with an average of 24 months (range from 6 to 43 months). All grafts were radiographically in good position at the time of follow-up. KT-1000 examination of affected knee showed less than 3 mm anterior translation difference compared with contralateral one's. Allograft group: 85 patients got normal IKDC score (81.7%). Lysholm score 82.8 +/- 8.5, Irgang score 79.2 +/- 7.3, Larson score 86.7 +/- 3.1. Autograft group: 29 patients got normal IKDC score (76.3%). Lysholm score 84.6 +/- 9.5, Irgang score 79.5 +/- 7.6, Larson score 88.9 +/- 6.8. No significant statistical difference was found between the 2 groups in the comprehend scale (P>0.05).

CONCLUSIONSBoth autograft and allograft group achieve good results, and the allograft B-PT-B could provide the similar clinical result as autograft, the preliminary result of allograft reconstruction might indicate predictable result in the future.

Adolescent ; Adult ; Anterior Cruciate Ligament ; surgery ; Anterior Cruciate Ligament Injuries ; Arthroscopy ; Bone-Patellar Tendon-Bone Grafting ; methods ; Female ; Follow-Up Studies ; Humans ; Male ; Matched-Pair Analysis ; Middle Aged ; Reconstructive Surgical Procedures ; methods ; Retrospective Studies ; Transplantation, Autologous ; Transplantation, Homologous ; Treatment Outcome

OBJECTIVETo label rat neural stem cells (NSCs) with the complex of Sinerem, the ultrasmall superparamagnetic iron oxide (USPIO), and poly-L-lysine (PLL), and evaluate the feasibility of tracking the labeled cells with magnetic resonance imaging (MRI) in vitro and in vivo.

METHODSSinerem was incubated with PLL to obtain the complex of Sinerem-PLL. The mesenchymal stem cells (MSCs) isolated from the bone marrow of SD rats were cultured and induced to differentiate into the neural stem cells. The second-passage cells were cultured overnight with the Sinerem-PLL complex, after which Prussian blue staining and transmission electron microscopy were performed to observe the nanoparticles in the cytoplasm. Cell apoptosis assay was performed to assess the cell viability 1 day, 1 week, and 2 weeks after the labeling. Cell tracking with 4.7 MR system was carried out in vivo and in vitro using T(2)WI and T(2)*WI sequences.

RESULTSThe NSCs could be effectively labeled with Sinerem-PLL complex with the labeling efficiency exceeding 95%. Prussian blue staining showed numerous blue iron particles in the cytoplasm, and under transmission electron microscope, these particles accumulated in the endosomes/lysosomes. The labeling did not significantly affect the cell viability and proliferation. Remarkable low signal density changes of the labeled cells was seen on T(2)WI and T(2)*WI in vivo and in vitro.

CONCLUSIONNSCs can be effectively labeled with Sinerem-PLL complex, and MRI can be used to track the labeled cells in vivo and in vitro.

Animals ; Cell Differentiation ; Cells, Cultured ; Dextrans ; metabolism ; Endosomes ; metabolism ; Ferrosoferric Oxide ; metabolism ; Lysosomes ; metabolism ; Magnetic Resonance Imaging ; methods ; Magnetite Nanoparticles ; Male ; Mesenchymal Stromal Cells ; cytology ; Microscopy, Electron, Transmission ; Neurons ; cytology ; metabolism ; ultrastructure ; Polylysine ; metabolism ; Rats ; Rats, Sprague-Dawley ; Stem Cells ; cytology ; metabolism ; ultrastructure ; Time Factors

Objective:To evaluate the residual stepping ability in monkeys with spinal cord injury longitudinally. Methods:Four adult female monkeys were studied. Right hemisection of 10 mm spinal cord tissue was performed at the T_7-9 segment. Gait tests of bipedal locomotion were performed before, and six weeks and twelve weeks after injury by VICON system. Gait cycle duration, amplitude of knee and ankle angles, and ratio of united parameters were obtained from successive stepping and were quantitative analyzed. Results:The coordination of bilateral hindlimbs was destroyed after spinal cord injury, and the right hindlimb showed obviously dragging. The gait cycle duration of the left hindlimb increased significantly (P < 0.001), and the amplitudes of knee and ankle angle significantly increased (P < 0.001) after spinal cord injury. The ratio of united parameters was not statistically different among all the time points (P > 0.05). The gait cycle duration of the left hindlimb was correlated with step length (r = 0.838, P = 0.001), step height (r = 0.726, P = 0.007) and amplitude of ankle angle (r = 0.766, P = 0.004), and the amplitude of ankle angle was correlated with step length (r = 0.627, P = 0.029). Conclusion:The gait pattern of monkey with spinal cord injury has been changed. The gait strategy of the uninjured side was adjusted compensatively after spinal cord injury to adapt the functional impairment of contralateral hindlimb.

The condition of patients with severe traumatic brain injury (sTBI) complicated by corona virus 2019 disease (COVID-19) is complex. sTBI can significantly increase the probability of COVID-19 developing into severe or critical stage, while COVID-19 can also increase the surgical risk of sTBI and the severity of postoperative lung lesions. There are many contradictions in the treatment process, which brings difficulties to the clinical treatment of such patients. Up to now, there are few clinical studies and therapeutic norms relevant to sTBI complicated by COVID-19. In order to standardize the clinical treatment of such patients, Critical Care Medicine Branch of China International Exchange and Promotive Association for Medical and Healthcare and Editorial Board of Chinese Journal of Trauma organized relevant experts to formulate the Chinese expert consensus on clinical treatment of adult patients with severe traumatic brain injury complicated by corona virus infection 2019 ( version 2023) based on the joint prevention and control mechanism scheme of the State Council and domestic and foreign literatures on sTBI and COVID-19 in the past 3 years of the international epidemic. Fifteen recommendations focused on emergency treatment, emergency surgery and comprehensive management were put forward to provide a guidance for the diagnosis and treatment of sTBI complicated by COVID-19.