Objective:To detect and verifica the gene profile difference of microRNA-146a (miR-146a) and its role in the pro-liferation of vascular smooth muscle cells (VSMCs) by gene chip technology. Methods: Artificially synthesized miR-146a mimics(50 nmol/L) ,miR-146 inhibitor ( 50 nmol/L ) , scramble ( 50 nmol/L ) and PBS were transfected into cultured primary rat VSMCs in vitro. After transfection,Real time PCR was used to measure the levels of miR-146a and the cell counting kit 8(CCK8) was employed to investigate the proliferation of VSMCs. The VSMCs interfered by miR-146a inhibitor or miR-146a control were examined by gene chips and the profile of gene were analyzed by bioinformatics technology to detect the different genes and signal transduction pathway. The changes in mRNAs and proteins were accessed separately by Real time PCR and Western blot. Results: Compared with sham and control VSMCs,miR-146a expression level was significantly decreased in treatment with miR-146a inhibitor(P<0. 01),as well as optical density(OD) was also shown remarkably down regulated simultaneously(P<0. 05). The investigation of gene profile revealed that the p53 signal pathway was up-regulated in VSMCs interfered by miR-146a. The mRNA and protein expression levels of p53, caspase3 and PTEN in p53 signal transduction pathway didn′t show significant differences(P>0. 05),however,the mRNA and protein expression levels of cyclin D1 significantly increased in treatment with miR-146a mimics VSMCs group and decreased in miR-146a inhibitor VSMCs group ( compared with sham VSMCs group, both P<0. 05 ) . Conclusion: Our data indicated that miR-146a may promote the proliferation of rat VSMCs by up-regulating cyclin D1 expression.

BACKGROUND:There is evidence that internal fixation through an anterior or posterior approach for treatment of severe kyphotic deformity in spinal tuberculosis exhibits good curative effects. However, few prospective, long-term fol ow-up case control studies are reported. OBJECTIVE:To investigate the efficacy and safety of cobalt al oy pedicle screw implantation for treatment of severe kyphotic deformity in spinal tuberculosis. METHODS/DESIGN:This is a prospective, single-center, self-control, open-label trial, which wil be performed at the Affiliated Hospital of Hebei University, China. Eighty-four patients with severe kyphotic deformity in spinal tuberculosis wil be included according to the diagnosis criteria. Among 52 patients with tuberculosis of the thoracic spine, 28 wil undergo surgery through a posterior approach, and 24 through an anterior approach. According to the Frankel Grade classification, grade C, D and E spinal cord function wil be assessed in 8, 31 and 13 patients, respectively. Surgery through a posterior and anterior approach wil be respectively performed in half of 32 patients with tuberculosis of the lumbar spine. Grade C, D and E spinal cord function wil be assessed in 7, 14 and 11 patients, respectively. The primary outcome measure of this study wil be the Cobb angle at the thoracic spine segments before and 2 years after surgery, which wil be used to evaluate the angle of the spine curvature at the thoracic segments. The secondary outcome measures wil be X-ray scan or MRI findings before and 2 years after surgery, which wil be used to evaluate vertebral fusion after internal fixation;and Frankel Grade before and 2 years after surgery, which wil be used to evaluate recovery of spinal cord function after injury. Other outcome measures wil include multiple logistic regression analysis results of the factors that influence patient's curative effects and the incidence of adverse events 2 years after surgery. The trial protocol has been approved by the Ethics Committee, Affiliated Hospital of Hebei University, China and wil be performed in strict accordance with the Declaration of Helsinki, formulated by the World Medical Association. Signed informed consent regarding the trial protocol wil be obtained from each participant. DISCUSSION:This study is to validate that cobalt al oy pedicle screw implantation shows precise curative effects in the treatment of severe kyphotic deformity in spinal tuberculosis and to analyze through what approach, posterior or anterior, internal fixation wil be more beneficial to surgery performance. The outcomes of this study wil provide objective long-term fol ow-up evidence for internal fixation treatment of severe kyphotic deformity in spinal tuberculosis in the clinic.

BACKGROUND: During the repair of thoracolumbar fracture, pedicle screw fixation is a commonly used treatment method. In the process of fixation, the different approaches can be used. OBJECTIVE: To compare effect and biocompatibility of pedicle screw by percutaneous approach, posterior median approach, and intervertebral space approach for thoracolumbar fracture. METHODS: 118 cases of thoracolumbar fracture were included after pedicle screw fixation. Al patients were divided into three groups according to the approach: posterior median approach group (38 cases), intervertebral space approach group (40 cases) and percutaneous approach group (40 cases). After 12 months of folow-up, perioperative conditions, pain score, vertebral height of anterior border, kyphosis correction effect, adverse events and biological compatibility were compared among three groups. RESULTS AND CONCLUSION:(1) Operation time, intraoperative bleeding and time in bed after surgery were shorter or less in the percutaneous approach and intervertebral space approach groups than in the posterior median approach group. Postoperative drainage was better in percutaneous approach and intervertebral space approach groups than in the posterior median approach group (alP < 0.05). Except drainage in the percutaneous approach and intervertebral space approach groups, no significant difference in other indicators was found. (2) Patients received imaging examination at different time points. The percentage of anterior vertebral height and kyphosis were significantly improved immediately after treatment and in final folow-up (alP < 0.05). No significant difference was detected before treatment, immediately after treatment and in final folow-up. (3) Visual Analogue score was identical before treatment. Visual analogue score was lower in the percutaneous approach and intervertebral space approach groups than in the posterior median approach group at 24 hours and 3 days after treatment and in final folow-up (alP < 0.05). No significant difference was detectable at 24 hours and 3 days after treatment and in final folow-up between the percutaneous approach and intervertebral space approach groups. (4) No rejection or wound non-healing was seen at 12 months after treatment. Some patients suffered from mild low back pain, which was improved by active symptomatic treatment. (5) These findings suggest that intervertebral space approach percutaneous approach obtained satisfactory outcomes compared with posterior median approach for treatment of thoracolumbar spine fractures, and good biocompatibility was found.

Objective: To understand the satisfaction of the residents in Hangzhou with primary health care services and its influencing factors, so as to provide basis for improving the quality of primary health care services. Methods: This study adopted multi-stage stratified random sampling method to recruit the residents aged 14 years or over in Hangzhou from July to September, 2019. A self-designed electronic questionnaire for primary health care services in Hangzhou was used to collect the demographic information, the utilization of primary health care institutions, the scores of the indicators of primary health care services and the satisfaction. The ordinal and multinomial logistic regression model was conducted to analyze the influencing factors for the satisfaction with primary health care services. Results: Totally 1 600 questionnaires were issued, 1 454 valid ones were collected, and the response rate was 90.88%. Five ( 0.34% ) residents were very dissatisfied, 16 ( 1.10% ) residents were dissatisfied, 153 ( 10.52% ) residents were generally satisfied, 780 ( 53.65% ) residents were satisfied, and 500 ( 34.39% ) residents were very satisfied with primary health care services. The ordinal and multinomial logistic regression analysis showed that basic medical insurance for urban employees ( OR=0.514, 95%CI: 0.370-0.714 ) , no health records in primary health institutions ( OR=0.528, 95%CI: 0.331-0.844 ) or unknown ( OR=0.524, 95%CI: 0.331-0.829 ) , the queuing time spent for getting medicines ( OR=0.698, 95%CI: 0.531-0.917 ) , ways of getting medicines (OR=0.639, 95%CI: 0.452-0.903 ) and completeness of testing equipment ( OR=1.315, 95%CI: 1.012-1.710) were influencing factors for the satisfaction with primary health care services. Conclusions The residents in Hangzhou are generally satisfied with the primary health care services, which are associated with the type of medical insurance, the condition of health records in primary institutions, the queuing time spent for getting medicines, the ways of getting medicines and the completeness of inspection equipment.

Bronchopulmonary dysplasia(BPD)is a common chronic respiratory disease, with high mortality and poor long-term prognosis in surviving children, and there is currently no effective intervention.Breastfeeding reduces the incidence of BPD.These depend on a variety of bioactive substances in human breast milk, such as immunoglobulins, oligosaccharides, exosomes, etc.Exosomes are nano-like vesicle bodies secreted by cells, which participate in information transmission and functional regulation between cells.They have high affinity for the body and can tolerate digestion of gastrointestinal enzymes and stomach acid.These enable them to target lung, liver and other organs smoothly.They are expected to become a new method for the prevention and treatment of BPD.

Objective : To analyze the status and influencing factors of the intention of people aged 18 to 25 years to primary medical institutions and their satisfaction for health services in Zhejiang Province, so as to provide basis for the improvement of health services in primary medical institutions. Methods: During November and December in 2019,the 18-25 year-old people in Zhejiang Province were recruited to investigate the general information, intention to seek medical advice and satisfaction for health services in primary medical institutions through WeChat. Logistic regression was performed to analyze the influencing factors of the intention and the satisfaction. Importance matrix was used to analyze the key drivers of the satisfaction. Results: Among the 620 people surveyed, with a response rate of 93.37%, 142 (22.90%) chose primary medical institutions for advice. Actually 516 (83.23%) people went to primary medical institutions last year, and 384 ( 74.42% ) of them were satisfied with the health services. The results of multivariate logistic regression analysis showed that the people aged 18-25 years who were under the new rural cooperative medical care system ( OR=3.062, 95%CI: 1.745-5.373 ) and who had records in community health centers ( OR=0.547, 95%CI: 0.308-0.970 ) were more likely to go to primary medical institutions for medical advice; the ability of doctors ( OR=1.478, 95%CI:1.168-1.871 ) ,the drug notification by medical staff ( OR=1.308, 95%CI: 1.065-1.606 ) , routine examination items ( OR=1.523, 95%CI: 1.227-1.889 ) , the ways of payment ( OR=1.168, 95%CI: 1.017-1.340 ) , the comfort of environment ( OR=1.785, 95%CI: 1.437-2.219 ) and the bulletin boards of health knowledge ( OR=1.302, 95%CI: 1.086-1.561 ) were associated with the satisfaction. The results of importance matrix analysis showed that the ability of doctors and routine examination items were the priorities to improve, followed by the drug notification by medical staff; the comfort of environment had competitive advantages; the ways of payment and the bulletin boards of health knowledge needed to be further analyzed. Conclusions The 18-25-year-old people in Zhejiang Province were less intended to seek medical advice in primary medical institutions, which was associated with the type of medical insurance and records in community health centers. They were satisfied with the health services, the ability of doctors and routine examination items were the key drivers.

Objective:To investigate effect of reperfusion injury following different ischemic duration on skeletal muscle in rats.Methods:A model of ischemia/reperfusion injury (IRI) was established by unilateral clamping femoral artery and additional application of tourniquet in skeletal muscle of hind limbs in 35 male Wsitar rats. According to different ischemia time, the animals were assigned to 2-hour ischemia and 24-hour reperfusion (I2R24 group), 2.5-hour ischemia and 24-hour reperfusion (I2.5R24 group), 3-hour ischemia and 24-hour reperfusion (I3R24 group), 4-hour ischemia and 24-hour reperfusion (I4R24 group) and sham group, with 7 rats per group. At the end of reperfusion, gastrocnemious tissues and plasma samples were collected and analyzed. The ratio of wet ∶ dry weight (W/D) was used to measure muscle edema. The assay of 3-(4, 5-Dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) was conducted to evaluate muscle viability. HE staining was executed to observe histopathological changes. Immunofluorescence staining was performed to assess the levels of C1q, C3b/c, tissue factor (TF), fibrinogen (FN), bradykinin receptor 1 (BR1), BR2, vascular cell adhesion molecule-1 (VCAM-1), E-selectin, fibrinogen-like protein-2 (FGL-2) and myeloperoxidase (MPO) in muscle tissues. ELISA method was used to determine the concentrations of interferonγ (IFN-γ), interleukin7 (IL-7), IL-18, macrophage inflammatory1α (MIP-1α) and monocyte chemotactic protein 1 (MCP-1) in plasma.Results:With prolongation of ischemia time and subsequent reperfusion, tissue edema became severe gradually. The ratio of W/D was 5.3±0.2, 6.1±0.3, 6.9±0.2, 7.6±0.3 in I2R24, I2.5R24, I3R24 and I4R24 groups, higher than that in sham group (4.5±0.1) (all P<0.01). Muscle viability got decreased gradually. Muscle viability was (62.4±3.5)%, (45.3±3.3)%, (35.4±3.4)%, (27.1±5.9)% in I2R24, I2.5R24, I3R24 and I4R24 groups, lower than that in sham group［(93.8±7.2)%］(all P<0.01). Histopathological changes became aggravated gradually. The most severe group was I4R24 group, with the most severe myocyte injury, interstitial edema and extensive inflammatory infiltration, followed by I3R24, I2.5R24 and I2R24 groups in order. There was normal structure integrity and neatly arranged myocyte in sham group. Meanwhile, levels of C1q, C3b, FN, BR1, VCAM-1, E-selectin and FGL-2 got increased gradually. The highest levels for these factors were seen in I4R24 group, followed by I3R24 group, I2.5R24 group, I2R24 group and sham group in order. The rough ratio of the number of positive MPO cells/total cell number under high lens (×200) were increased gradually, with the highest level in I4R24 group, followed by I3R24 group, I2.5R24 group, I2R24 group and sham group in order. However, expression of TF and BR2 were not altered significantly among the groups. Plasma levels of INF-γ, IL-7, IL-18, MIP-1α and MCP-1 elevated gradually with prolongation of ischemia time (all P<0.01). The sequence was the sham group, I2R24 group, I2.5R24 group, I3R24 group and I4R24 groups for levels of these factors from low to high (all P<0.01). Conclusion:Reperfusion after prolongation of ischemia duration can increase the activation of complement, coagulation, kinin and endothelial cells as well as the release of inflammatory factors, and thus aggravate the degree of skeletal muscle tissue injury.

Breast milk contains a variety of bioactive components, which play a vital role in disease prevention and treatment.Among them, circular RNA(circRNA), which is a closed ring structure formed by covalent bonds, has aroused interest because of its conservation and stability.Pediatric researchers should attach importance to studies on circRNA in breast milk, as they may bring new inspiration for breast milk functions.In this article, characteristics and functions of circRNA in breast milk will be investigated, and its research and prospects will be discussed.

Objective　To dissect the clinical and imaging differences between atypical multiple sclerosis and cerebral small vessels diseases.Methods　We presented one case of a hereditary cerebral small vessel disease-autosomal dominant genetic disease combined with subcortical infarction and white matter encephalopathy (CADASIL),which was initially misdiagnosed as multiple sclerosis.Afterwards.We reviewed literature regarding typical images and clinical aspects in multiple sclerosis and cerebral small vessels diseases and discussed the differential diagnosis points.Results　Multiple sclerosis often starts in young adults,with IgG index greater than 0.7,positive oligoclonal band,central venous signs,few microbleeding and asymmetric periventricular white matter lesions on MRI scan.On the contrary,cerebral small vessel diseases often start in old patients with vascular risk factors.MRI scan often shows the lacunar cerebral infarction,microbleeding,brain atrophy and symmetric periventricular white matter hyperintensity.Conclusion　Understanding the differential points between the two diseases may help avoid misdiagnosis and initiate reasonable treatment immediately.

Currently,human health due to corona virus disease 2019(COVID-19)pandemic has been seriously threatened.The coronavirus severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)spike(S)protein plays a crucial role in virus transmission and several S-based therapeutic approaches have been approved for the treatment of COVID-19.However,the efficacy is compromised by the SARS-CoV-2 evolvement and mutation.Here we report the SARS-CoV-2 S protein receptor-binding domain(RBD)inhibitor licorice-saponin A3(A3)could widely inhibit RBD of SARS-CoV-2 variants,including Beta,Delta,and Omicron BA.1,XBB and BQ1.1.Furthermore,A3 could potently inhibit SARS-CoV-2 Omicron virus in Vero E6 cells,with EC50 of 1.016 pM.The mechanism was related to binding with Y453 of RBD deter-mined by hydrogen-deuterium exchange mass spectrometry(HDX-MS)analysis combined with quan-tum mechanics/molecular mechanics(QM/MM)simulations.Interestingly,phosphoproteomics analysis and multi fluorescent immunohistochemistry(mIHC)respectively indicated that A3 also inhibits host inflammation by directly modulating the JNK and p38 mitogen-activated protein kinase(MAPK)path-ways and rebalancing the corresponding immune dysregulation.This work supports A3 as a promising broad-spectrum small molecule drug candidate for COVID-19.