【Abstract】 Objective To isolate and culture human amniotic fluid-derived mesenchymal stem cells (HAFMSCs),to investigate a better cryopreservation protocol of HAFMSCs and to observe the biocharacteristics and the multi-potential of HAFMSCs after cryopreservation for the further fundamental researches and cl inical appl ications. Methods HAFMSCswere isolated from the amniotic fluid of pregnant women during the second trimester by the improved two-step method.HAFMSCs were cryopreserved with different cryopreservation protocols (containing different contents of FBS and DMSO atcryoprotectant) in l iquid nitrogen for 12 weeks. The biocharacteristics of the HAFMSCs after cryopreservation were analyzed. The growth characteristics were observed by MTT method and the growth curves were drawn. The surface antigens of HAFMSCs were detected using flow cytometry, including CD29, CD34, CD44, CD45, CD73, and CD90. The adi pogenic and osteogenic differentiation abil ities of HAFMSCs were observed. The mRNA levels of Oct-4 and Nanog of the HAFMSCs were compared between before and after cryopreservations. Results At 12 weeks after cryopreservation, different protocols had different effects on the cell viabil ity; the better formula of cryoprotectant was 50% DMEM, 40% FBS, and 10% DMSO. After cryopreservation, the cells proliferated rapidly and the growth curves showed “S” shape, which was the same as the cells before cryopreservation. Phenotype showed that HAFMSCs were positive for the surface markers CD29, CD44, CD73, and CD90, and negative for CD34 and CD45. After 21 days of adi pogenic differentiation, the l ipid droplets were observed by oil red O staining. After 21 days of osteogenic differentiation, the calcium mineralizations were verified by von Kossa staining. There was no significant difference (P gt; 0.05) in the mRNA levels of Oct-4 and Nanog between before and after cryopreservations. Conclusion HAFMSCs have rapid proliferation and multi-potential in vitro. The cells have high viabil ities and no changes of the biocharacteristics and differentiation potential ities after cryopreservation for 12 weeks. Cryoprotectant containing 50% DMEM, 40% FBS, and 10% DMSO is a better cryopreservation protocol.
Electroencephalogram (EEG) is the primary tool in investigation of the brain science. It is necessary to carry out a deepgoing study into the characteristics and information hidden in EEGs to meet the needs of the clinical research. In this paper, we present a wavelet-nonlinear dynamic methodology for analysis of nonlinear characteristic of EEGs and delta, theta, alpha, and beta sub-bands. We therefore studied the effectiveness of correlation dimension (CD), largest Lyapunov exponen, and approximate entropy (ApEn) in differentiation between the interictal EEG and ictal EEG based on statistical significance of the differences. The results showed that the nonlinear dynamic characteristic of EEG and EEG subbands could be used as effective identification statistics in detecting seizures.
ObjectiveTo analyze the clinical and epidemiological characteristics of hospitalized avian influenza A (H7N9) virus infections in Hunan province from 2013 to 2017, and provide evidences for control, diagnosis and treatment of this disease.MethodsNinety-one hospitalized patients were confirmed with H7N9 infection in Hunan. Excluding 2 patients less than 18 years old and 10 with missing data, 79 patients with H7N9 infection were analyzed.ResultsMost confirmed cases were affected in the second and fifth epidemic wave and number of patients in the fifth wave was more than the sum in prior 4 waves. Epidemiological characteristics, clinical symptoms and case fatality did not change significantly. Administration of antiviral drugs was more active in the fifth wave [from illness onset to antiviral drug: (6.3±2.4)d vs. (7.6±2.4)d, P=0.047]. Multiple logistic regression analysis showed that shock (OR=4.683, 95%CI 1.136–19.301, P=0.033) was the independent risk factor of H7N9 infections. There were no significant differences in case fatality among group oseltamivir, group oseltamivir+peramivir, and group peramivir.ConclusionsPatients with avian influenza A (H7N9) increased in the fifth wave but clinical characteristics changed little. Antiviral treatment should be more active. Shock is an independent risk factor of H7N9 infections. Oseltamivir-peramivir biotherapy can not reduce case fatality compared with oseltamivir or peramivir monotherapy.
ObjectiveTo analyze the microbiological characteristics of airway bacteria in adult patients with bronchiectasis and to analyze their correlation with the clinical features. MethodsPatients diagnosed with bronchiectasis in the Department of Respiratory and Critical Care Medicine of West China Hospital of Sichuan University from October 2017 to April 2018 were classified into the bronchiectasis group, while the control group was those who were found to have pulmonary nodules (diameter less than 10 mm) requiring bronchoscopy by physical examination. All subjects in both groups had not used antibiotics or hormones within 4 weeks and had no other respiratory diseases. Bronchoalveolar lavage fluid (BALF) from the lesion site of the branchial expansion group was collected, and BALF from the basal segment of the contralateral inferior bronchial lobe of the pulmonary nodule was collected in the control group. Bacterial culture and 16S rRNA gene sequencing were performed in both groups. ResultsSeventeen cases and six controls were enrolled in this study and the BALF specimens were collected. Eight cases were in stable period and nine cases were in acute period. The case group was divided into the bacteria-positive group and negative group based on bacterial culture of BALF. Shannon index in the bacteria-positive group was significantly lower than the bacteria-negative group and the control group. And Shannon index showed a negative correlation with positive bacterial culture in BALF. When Shannon index ≤4.5 was used to predict positive bacterial culture, the sensitivity and specificity were 83.3% and 90.9% respectively. The average relative abundance of bacteria was higher and the average sample distribution uniformity was lower in patients with acute period, compared with those in patients with stable period. Shannon index was negatively correlated with the acute exacerbation in patients. When Shannon index <5.0 was used to predict acute exacerbation, the sensitivity and specificity were 77.8% and 100.0%, respectively. ConclusionsShannon index in 16S rRNA gene sequencing results has certain predictive value for acute exacerbation stage. 16S rRNA gene sequencing combined with bacterial culture results can help guide clinicians to provide more precise treatment plans.
ObjectiveTo establish the model of compression fracture of acetabular dome, and to measure the contact characteristics of acetabular weight-bearing area of acetabulum after 3 kinds of internal fixation.MethodsSixteen fresh adult half pelvis specimens were randomly divided into 4 groups, 4 specimens each group. Group D was the complete acetabulum (control group), and the remaining 3 groups were prepared acetabular dome compression fracture model. The fractures were fixed with reconstruction plate in group A, antegrade raft screws in group B, and retrograde raft screws in group C. The pressure sensitive films were attached to the femoral head, and the axial compression test was carried out on the inverted single leg standing position. The weight-bearing area, average stress, and peak stress were measured in each group.ResultsUnder the loading of 500 N, the acetabular weight-bearing area was significantly higher in group D than in other 3 groups (P<0.05), and the average stress and peak stress were significantly lower than in other 3 groups (P<0.05). The acetabular weight-bearing area were significantly higher in group B and group C than in group A, and the average stress and peak stress were significantly lower than in group A (P<0.05). There was no significant difference in the above indexes between group B and group C (P>0.05).ConclusionFor the compression fracture of the acetabular dome, the contact characteristics of the weight-bearing area can not restore to the normal level, even if the anatomical reduction and rigid internal fixation were performed; compared with the reconstruction plate fixation, antegrade and retrograde raft screws fixations can increase the weight-bearing area, reduce the average stress and peak stress, and reduce the incidence of traumatic arthritis.
Objective
To study relation between expression of cyclin-dependent kinase like 5 (CDKL5) in gastric cancer tissue and clinicopathologic characteristics of patients with gastric cancer.
Methods
The expressions of CDKL5 mRNA and protein in 45 gastric cancer tissues and their corresponding adjacent tissues were detected by real-time quantitative PCR and Western blot methods respectively. Meanwhile the expressions of CDKL5 protein in gastric cancer cell lines including AGS and MKN-45 were detected by immunofluorescence method.
Results
The CDKL5 mRNA and protein highly expressed in the 45 primary gastric cancer tissues as compared with the their corresponding adjacent tissues (P<0.05). The results of univariate analysis identified that the age (P=0.033), vascular invasion (P=0.007), T stage (P=0.049), and TNM stage (P=0.041) were associated with the expression of CDKL5 mRNA in the primary gastric cancer tissues. The results of multivariate regression analysis showed that the vascular invasion (P=0.013) and TNM stage (P=0.024) were the important factors affecting the expression of CDKL5 in the primary gastric cancer tissues. The CDKL5 protein expressions were found at fragments with relative molecular masses of 107×103 and 85×103 in the primary gastric cancer tissues and gastric cancer cell lines.
Conclusions
CDKL5 mRNA and protein highly express in primary gastric cancer tissue and relate to vascular invasion and TNM stage. The CDKL5 protein expresses at fragments with relative molecular masses of 107×103 and 85×103 in primary gastric cancer tissues and gastric cancer cell lines suggest that CDKL5 has different expressing way in protein level.
ObjectiveTo systematicly evaluate expression of epithelial cell adhesion molecule (EpCAM) in colorectal cancer (CRC) and its correlation with clinicopathologic characteristics of patient with CRC.MethodsPubMed, Web of Science, Cochrane Library, Embase, CNKI, Wanfang, VIP, and other databases were searched comprehensively. The retrieved literatures were imported into Endnote X9. The data about the expression of EpCAM in the CRC and the relationship between EpCAM expression and clinicopathologic characteristics of patients with CRC were screened and extracted. RevMan 5.3 software was used for meta-analysis.ResultsA total of 5 396 patients with CRC were included. The meta-analysis results showed that the expression rate of EpCAM in the CRC tissues or blood was significantly higher than that in the benign colorectal tumor and normal tissue or blood (P<0.05). The high expression rates of EpCAM in the Dukes C+D stage, tumor diameter >3 cm, infiltration state of tumor margin, with lymph node and distant metastasis of the CRC were significantly higher than those in the A+B stage, tumor diameter ≤3 cm, dilated state of tumor margin, without lymph node and distant metastasis (P<0.05).ConclusionResults of this meta-analysis suggest that expression of EpCAM might be related to some clinicopathologic characteristics (carcinogenesis, Dukes stage, tumor size, tumor margin morphology, lymph node metastasis, distant metastasis) of patients with CRC.
Objective To analyze the characteristics of femoral neck fractures in young and middle-aged adults by means of medical image analysis and fracture mapping technology to provide reference for fracture treatment. Methods A clinical data of 159 young and middle-aged patients with femoral neck fractures who were admitted between December 2018 and July 2019 was analyzed. Among them, 99 patients were male and 60 were female. The age ranged from 18 to 60 years, with an average age of 47.9 years. There were 77 cases of left femoral neck fractures and 82 cases of right sides. Based on preoperative X-ray film and CT, the fracture morphology was observed and classified according to the Garden classification standard and Pauwels’ angle, respectively. Mimics19.0 software was used to reconstruct the three-dimensional models of femoral neck fracture, measure the angle between the fracture plane and the sagittal plane of the human body, and observe whether there was any defect at the fracture end and its position on the fracture surface. Through reconstruction, virtual reduction, and image overlay, the fracture map was established to observe the fracture line and distribution. Results According to Garden classification standard, there were 6 cases of type Ⅰ, 61 cases of type Ⅱ, 54 cases of type Ⅲ, and 38 cases of type Ⅳ. According to the Pauwels’ angle, there were 12 cases of abduction type, 78 cases of intermediate type, and 69 cases of adduction type. The angle between fracture plane and sagittal plane of the human body ranged from –39° to +30°. Most of them were Garden type Ⅱ, Ⅳ and Pauwels intermediate type. The fracture blocks were mainly in the form of a triangle with a long base and mainly distributed below the femoral head and neck junction area. Twenty-six cases (16.35%) were complicated with bone defects, which were mostly found in Garden type Ⅲ, Ⅳ, and Pauwels intermediate type, located at the back of femoral neck and mostly involved 2-4 quadrants. The fracture map showed that the fracture line of the femoral neck was distributed annularly along the femoral head and neck junction. The fracture line was dense above the femoral neck and scattered below, involving the femoral calcar. Conclusion The proportion of displaced fractures (Garden type Ⅲ, Ⅳ) and unstable fractures (Pauwels intermediate type, adduction type) is high in femoral neck fractures in young and middle-aged adults, and comminuted fractures and bone defects further increase the difficulty of treatment. In clinical practice, it is necessary to choose treatment plan according to fracture characteristics. Anatomic reduction and effective fixation are the primary principles for the treatment of femoral neck fracture in young and middle-aged adults.
ObjectiveTo investigate the clinical characteristics of epileptics with pregnancy and then provide reference for standardized management of epileptics with pregnancy. MethodsFrom June 2012 to June 2021, epileptics with pregnancy who delivered in Jinan Central Hospital were selected as the research subjects. The clinical data such as the application of Antiseizure medications (ASMs) during pregnancy, seizure frequency, pregnancy outcomes, delivery ways, offspring feeding ways and the incidence of complications were investigated and analyzed. ResultsAmong 36 epileptics with pregnancy, 20 cases (55.56%) were treated with ASMs alone, 5 cases (13.88%) were treated with combined medication, and 11 cases (30.56%) were treated without ASMs during pregnancy. 15 cases (41.67%) adhered to systematic application of ASMs, 17 cases (47.22%) did not adhere to systematic application of ASMs, and 4 cases (11.11%) had unknown medication history. The frequency of seizures increased in 5 cases, decreased in 7 cases and unchanged in 24 cases during pregnancy. Pregnancy outcomes: full-term delivery in 33 cases (91.67%), preterm delivery in 1 case (2.78%) and abortion in 2 cases (5.56%). Delivery mode: cesarean section in 31 cases (91.18%), vaginal delivery in 3 cases (8.82%). After delivery, 4 cases (11.76%) were fed with milk powder and 30 cases (88.24%) were breast-fed. Complications: There were 6 cases complicated with anemia (16.67%), 5 cases complicated with gestational hypertension (13.89%), 3 cases complicated with gestational diabetes (8.33%), 4 cases complicated with premature rupture of membranes (11.11%), 2 cases complicated with fetal growth restriction (5.56%), 2 cases complicated with oligohydramnios (5.56%), 3 cases complicated with fetal distress (8.33%) and 3 cases complicated with neonatal asphyxia (8.33%). ConclusionsThe proportion of epileptics with pregnancy who were systematically treated with ASMs was low and the seizures were poorly controlled. There is a lack of standardized management for such patients in clinical practice.
The aim of this study is to explore the effects of continuous theta-burst transcranial magnetic stimulation (cTBS) on functional brain network in emotion processing. Before and after the intervention of cTBS over left dorsolateral prefrontal cortex (DLPFC) of ten participants who were asked to perform the emotion gender recognition task, we recorded their scalp electroencephalograms (EEG). Then we used the phase synchronization of EEG to measure the connectivity between two nodes. We then calculated the network efficiency to describe the efficiency of information transmission in brain regions. Our research showed that after the intervention of cTBS and the stimulation of the emotion face picture, there was an obvious enhancement in the event-related spectral perturbation after stimuli onset in beta band in 100–300 ms. Under the stimulation of different emotion picture, the values of global phase synchronization for negative and neutral stimuli were enhanced compared to positive ones. And the increased small-worldness was found in emotional processing. In summary, based on the effect of activity change in the left DLPFC on emotion processing brain network, the emotional processing mechanism of brain networks were preliminary explored and it provided the reference for the research of emotion processing brain network in the future.
