Rotating wall vessels (RWVS), an ingenious apparatus for three-dimensional suspension culture, is widely used to build a simulated microgravity-effect on cell. Independent researchers have proposed hypotheses to illustrate why RWVS can simulate certain aspects of microgravity. Many of the hypotheses stated that the culture condition in RWVS is determined by the cellular mechanical environment which is a result of low fluid shear and microcarrier's motion. The microcarrier's motions consist of primary and secondary motions. In the light of the analysis of forces loaded by the microcarriers, some conclusions are drawn from the data on microcarriers' primary and secondary motions about which many simulations and observations have already been conducted.
Cell Culture Techniques ; instrumentation ; methods ; Gravitation ; Models, Theoretical ; Motion ; Rotation ; Stress, Mechanical ; Weightlessness Simulation

Objective To study features of the MRI and clinic in a family with pure hereditary spastic paraplegia (PHSPG) type 6.Methods Target loci (SPG3, 4, 6, 8 10 and 12) linkage analysis was performed in a SPG pedigree having 6 affected individuals using microsatellite markers and NIPA1 gene was screened for mutation by PCR-amplification and sequencing. MRI of brain and cervical and thoracic spinal cord were examined in these 6 patients and 6 normal controls matched for age and sex by two independent radiologists blinded to the clinical diagnosis. Cross-sectional areas and anteroposterior and transverse diameters of the spinal cord at the levels of C2～3, C7, T1～4, T9 were measured and data was statistically analyzed using the student's t test. Results A missense mutation of 316g→c in NIPA1 was identified in the affected subjects, presumably resulting in substitution of glutamic acid for arginine in residue 106. Evaluation of the brain MRI images revealed non-specific brain abnormalities. All patients presented thinning of cervical and upper thoracic spine with atrophy in both gray and white matter and enlarged subarachnoid cavity. In severe atrophic segments, a distinct boundary between grey and white matter was observed and the lesions in grey matter presented literal high intensity spots or patches with clear boundary on transaxial T2-weighted images (T2WI) and high signal intensity longitudinal strip on the sagittal T2WI. Cross-sectional areas and anteroposterior and transverse diameters of the spinal cord at C2～3, C7, T1～4 were significantly smaller in patients than in controls, while at the T9 level only transverse diameter showed significant difference (7.22±0.08 vs 8.17±0.41, t=2.870, P=0.046). Conclusions These findings indicate that the disease process in patients with SPG6 might be confined to the cervical and thoracic spinal cord, with atrophy in both white and grey matter having a distinct boundary.

Objective To modify anterior cruciate ligament (ACL)reconstruction to achieve better structure;to measure the tunnel's position and shape using three-dimensional computed tomography (CT)so as to show the change of the femoral bony tunnel.Methods Forty-five patients were diagnosed with an ACL tear and subsequently underwent the modified ACL reconstruction,using a minor-diameter drill to drill the femoral tunnel and then expand it to oval-shaped tunnel with the raspatory between July 2014 and 2015.All procedures were conducted by the same experienced surgeon (Ao Yf.).In all patients,CT was performed at one week after the ACL reconstruction to evaluate the femoral tunnel's positioning,then the CT-images were imported into the image processing software MIMICS to reconstruct the femoral bony tunnel to measure the tunnel's diameters of the section and the shape.The tunnel's position was evaluated using the quadrant method.Results In all the 45 patients,the graft could pass the tunnel.And no posterior tunnel wall blowout,neurological or vascular deficit was documented during the operation.According to the different methods of modification (6 to 7,6 to 8 and 7 to 8),the long diameters of the sections of the femoral bony tunnels were 8.34-± 0.52 mm,10.65 ± 0.72mm,9.27-± 0.11 mm,without significant difference from the theoretical value (8.16 mm,10.66 mm and 9.14 mm).However,the short diameters of the sections were 6.57--0.12 mm,6.74-± 0.13 mm,and 7.52 ± 0.05 mm,showing significant difference from the theoretical value (6mm,6mm and 7 mm).The average centre of the femoral tunnel was located at 26.4 ± 4.8％ of the femoral length and at 30.1 ± 5.6％ of the femoral height.Conclusion In this modified ACL reconstruction,a minor-diameter drill is used to drill the femoral tunnel and then expand it to oval-shaped tunnel with the raspatory,The graft was matched well with the bony tunnel intraoperatively.The three-dimensional CT shows that the femoral tunnels are well located within the anatomical ACL footprint,and its entrance is closer to the natural state.

This study sought to detect the pathological changes of anterior cruciate ligament (ACL) and medial collateral ligament (MCL) under injury stretch. Bone-ACL-Bone (B-ACL-B) and B-MCL-B complexes were isolated from 20 male Wister rats, and were immersed in phosphate buffered saline. The complexes were stretched with 10% or 20% strain for 10 min or 30 min. After being stretched, the specimens were fixed in 10% buffered formalin, then mounted in paraffin. Sections were stained with Alcian blue-PAS and HE. The following results were found: In the control group, the matrix in ACL contained much more GAGs, as compared with that in MCL. When stretched with 10%, most of the fibroblasts in ACL were elongated like spindles in shape, and some pyknotic nuclei were found increased with stretching time. With 20% strain, ACL showed disruption in parts of collagen fibrils and lysis. But MCL was often torn at its tibia end. The injury can be detected in pathological slices under microscope, even this injury can not be found with naked eye. This injury first starts with the disturbance of the nucleus in the ligament, but following further stretching, it will extend to the rupture of collagen fibrils, and the serious injury of the fibroblasts is especially bad to the repair of the ligament.
Animals ; Anterior Cruciate Ligament ; pathology ; Anterior Cruciate Ligament Injuries ; Male ; Medial Collateral Ligament, Knee ; injuries ; pathology ; Rats ; Rats, Wistar ; Stress, Mechanical

Objective To compare the graft maturity of the anterior cruciate ligament reconstruction using the oval-shaped tunnel technique and round-shaped tunnel technique by signal/noise quotient (SNQ) of MRI postoperatively.Methods Forty patients diagnosed as the anterior cruciate ligament rupture between August 2015 and June 2016 were included according to the inclusion and exclusion criteria.Patients were randomized into a round-shaped group and an oval-shaped group,undergoing traditional round-shaped tunnel and oval-shaped tunnel reconstruction of the anterior cruciate ligament respectively.One year postoperatively,the MRI was conducted,and three intra-articular regions of interest (ROI) were selected to compare the graft maturity by calculating SNQ.Results None of the forty patients experienced complications of bone tunnel blowout,graft getting through difficulties and neurological or vascular injuries.SNQ of the round-shaped group were 3.72 ± 2.29,significantly higher than that of the oval-shaped group(P＜0.001).Moreover,SNQ of proximal ROI and distal ROI of ovalshaped group were 1.97 ± 1.30 and 2.76 ± 1.75,significantly lower than the round-shaped group with proximal site of 3.53 ± 2.11(P=0.008) and distal site of 4.46 ± 2.28(P=0.012).Conclusion Comparing MRI signal intensity one year after the treatment,we have found the graft SNQ after oval-shaped tunnel reconstruction was lower than the round-shaped reconstruction,with better graft maturity.

Objective To explore the role of β-1,3-galactosyltransferase 2(B3galt2)in mice with cerebral ischemic injury.Methods Adult male C57BL/6J mice were randomly divided into the sham,suture-occluded middle cerebral artery occlusion(MCAO)model,MCAO model+lentiviral vector control(LV-GFP),and MCAO model+lentiviral vector overexpression B3galt2(LV-B3galt2)groups,with six mice in each group.Neurological deficit scoring and rotating rod experiments were performed 24 h after ischemia in each group,and 2,3,5-triphenyltetrazolium chloride(TTC)staining was used to determine the infarction volume.The number of neurons in the ischemic cerebral cortex was determined in each group using Nissl staining.The levels of oxidative stress-related factors in the brain tissues were detected using the relevant kits.Results Compared with the sham group,the MCAO model group showed increased infarct volume and neurological deficits(P<0.05),significantly decreased number of neurons in the ischemic cerebral cortex and levels of super-oxide dismutase(SOD)and glutathione peroxidase(GSH)(all P<0.05),and significantly increased levels of reactive oxygen species(ROS)and malondialdehyde(MDA)(all P<0.05).Compared with the MCAO model group,the LV-B3galt2 group had reduced volume of cerebral infarction,significantly improved neurological deficits(all P<0.05),significantly increased number of neurons in the ischemic cerebral cortex of mice,significantly decreased levels of ROS and MDA(P<0.05),and significantly elevated levels of SOD and GSH(all P<0.05).Conclusion B3galt2 overexpression can reduce brain injury in an ischemic damage mouse model,and its mechanism may be through the inhibition of oxidative stress reactions.

OBJECTIVETo investigate the factors which may influence the imatinib plasma concentration in Chinese patients with gastrointestinal stromal tumor(GIST), and to illuminate the significance of monitoring imatinib plasma concentration in adjuvant therapy for patients with GIST.

METHODSA cross-sectional study with 60 GIST patients who accepted the imatinib therapy after surgery was conducted. They were respectively administrated in 10 domestic hospitals from December 2014 to April 2016, including The First Affiliated Hospital of Nanjing Medical University(n=28), The Affiliated Hospital of Nantong University(n=9), The Affiliated Hospital of Xuzhou Medical College(n=6), Nanjing Drum Tower Hospital(n=5), The Second Affiliated Hospital of Nanjing Medical University (n=2), Jingling Hospital (n=2), The Second People's Hospital of Lianyungang(n=2), Shandong Provincial Hospital(n=2), Jiangsu Province Tumor Hospital(n=2), and The First Affiliated Hospital of Zhejiang University(n=2). Some specific time points for collecting blood sample before and after taking imatinib were determined, then liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was used for monitoring imatinib plasma concentration in patients with GIST. Linear regression analysis was used for the correlation analysis of imatinib plasma concentration with dosage, clinicopathologic feature and side effect.

RESULTSPatients who could not tolerate 400 mg imatinib per day(n=3) received 300 mg per day. There was no significant difference in imatinib plasma concentration between patients with 300 mg and those with 400 mg imatinib(n=53)(P=0.527). However, the imatinib plasma concentration in patients with 600 mg imatinib per day (n=4) was significantly higher as compared to those with 400 mg(P=0.000). Linear regression analysis indicated a negative correlation between the imatinib plasma concentration in patients with 400mg imatinib per day for 90 days continuously and body surface area(R=0.074, P=0.035), but no significant correlations of with age, creatinine clearance and serum albumin concentration were observed (all P>0.05). The differences in imatinib plasma concentration were not statistically significant between patients of different gender and those taking proton-pump inhibitor (PPI) or not (both P>0.05). Difference in imatinib plasma concentration between patients with different surgery was significant (P=0.026). Compared to patients who underwent wedge resection, enterectomy and other surgeries, the imatinib plasma concentration of patients with subtotal gastrectomy or total gastrectomy decreased significantly (all P<0.05). After 90 days of taking imatinib continuously, linear regression analysis revealed a negative correlation between imatinib plasma concentration in patients with 400 mg imatinib per day and white blood cell count (R=0.103, P=0.013), and a positive correlation with serum alanine aminotransferase (ALT) concentration (R=0.076, P=0.033).

CONCLUSIONSThe imatinib plasma concentration in patients with larger body surface area, subtotal gastrectomy or total gastrectomy may be lower. For these patients, dosage of imatinib should be considered to increase in order to achieve effective plasma concentration. Excessive imatinib plasma concentration can result in some side effects, such as decrease of white blood cells and liver damage. Therefore, it is significant for receiving optimal clinical therapeutic efficacy to monitor imatinib plasma concentration, adjust imatinib dosage timely and keep imatinib plasma concentration in effective and safe range.

Adult ; Antineoplastic Agents ; administration & dosage ; pharmacokinetics ; Benzamides ; Combined Modality Therapy ; Cross-Sectional Studies ; Female ; Gastrectomy ; Gastrointestinal Stromal Tumors ; drug therapy ; surgery ; Humans ; Imatinib Mesylate ; administration & dosage ; pharmacokinetics ; Male ; Middle Aged ; Piperazines ; Pyrimidines ; Tandem Mass Spectrometry