ObjectiveTo explore the mechanism of Tibetan medicine Ershiwuwei Guijiuwan against osteoporosis in ovariectomized rats through the classical Wnt/β-catenin signaling pathway. MethodForty-eight 3-month-old SD female rats were randomized into model group （equivalent volume of normal saline）， sham operation group （equivalent volume of normal saline）， estradiol group （0.1 mg·kg^-1 estradiol valerate）， and high-， medium-， low-dose Ershiwuwei Guijiuwan groups （800， 400， 200 mg·kg^-1， respectively）. For the modeling， some adipose tissue near the ovaries was removed in the sham operation group， and ovaries were excised in other groups. Administration （ig， once a day） started two weeks after the operation and lasted 12 weeks. After sampling， based on hematoxylin and eosin （HE） staining， the morphological changes of the right femur and lumbar spine of the rats were observed. The enzyme-linked immunosorbent assay （ELISA） was performed to detect the serum levels of rat tartrate-resistant acid phosphatase 5b （TRAP5b）， collagen type Ⅰ C-terminal peptide （CTX-Ⅰ）， bone alkaline phosphatase （BALP）， amino-terminal propeptide of type Ⅰ procollagen （PINP）， and bone Gla-protein （BGP）. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was performed to examine the mRNA expression of β-catenin and Runt-related transcription factor 2 （Runx2） in femur， and Western blot to detect the protein expression of β-catenin， Runx2， and low-density lipoprotein receptor-related protein-5 （Lrp-5）. ResultCompared with the sham operation group， the model group showed disordered and sparse bone trabecula with fewer connections， decrease in serum levels of BALP， BGP， and PINP （P<0.01）， increase in levels of CTX-Ⅰ and TRAP5b （P<0.01）， reduction in mRNA expression of β-catenin and Runx2 in femoral tissue （P<0.01） and protein expression of Lrp-5， β-catenin， and Runx2 （P<0.01）. Compared with the model group， estradiol and each dose of Ershiwuwei Guijiuwan increased the volume of bone trabecula， made thebone trabecula closely arranged， reduced the loss of the trabecular meshwork， raised the serum levels of BALP and BGP （P<0.01）， and lowered TRAP5b level （P<0.01）. PINP level was significantly increased in the high-dose Ershiwuwei Guijiuwan group and estradiol group （P<0.01） and CTX-Ⅰ level was significantly decreased in the high-dose Erwenwuwei Guijiuwan group and the estradiol group （P<0.01） compared with those in the model group. The mRNA expression of β-catenin in femoral tissue and protein expression of Lrp-5 and β-catenin in estradiol group and three Ershiwuwei Guijiuwan groups were significantly increased （P<0.05， P<0.01）， and the levels of Runx2 mRNA and protein were significantly increased in the high-dose Ershiwuwei Guijiuwan group and the estradiol group （P<0.01） compared with those in the model group. ConclusionTibetan medicine Ershiwuwei Guijiuwan is effective for the osteoporosis in ovariectomized rats， which may be related to the classic Wnt/β-catenin signaling pathway. It affects bone metabolism by up-regulating the expression of osteogenesis-related genes in the signaling pathway.

Objective:To investigate the role and probable mechanism of miRNA-181a in nonalcoholic fatty liver disease.Methods:HepG2 cells were treated with palmitic acid to construct a nonalcoholic fatty liver disease cell model, and the expression of miR-181a and lipidosis in the cells were measured. Transforming growth factor-β (TGF-β) was used to examine the effect of miR-181a expression in HepG2 cells. The miR-181a, lipidosis, reduced glutathione and reactive oxygen species (ROS) were determined by controlling and regulating the miR-183 expression levels after transfection with miR-181 mimics and inhibitors in HepG2 cells. The miR-181a target genes were predicted by bioinformatics analysis, and verified by real-time fluorescent quantitative PCR and western blotting. The independent sample t-test was used for the comparison between the two independent samples, and the comparison between multiple groups were accorded with the normal distribution, homogeneity of variance, and one-way analysis of variance.Results:Lipidosis was significantly increased after palmitic acid treatment in HepG2 cells, and the expression level of miR-181a was significantly increased than control group. After HepG2 cells were transfected with miR-181a inhibitors, the expression of miR-181a, triglycerides and reactive oxygen species were down-regulated, and reduced glutathione, predicting the mRNA and protein expression of target gene silencing information regulator 2 related enzyme 1 were up-regulated. However, the results were contrary to the above changes after transfection with miR-181a mimics.Conclusion:miR-181a participates in lipidosis and promotes lipid peroxidation in nonalcoholic fatty liver disease. miR-181a may affect the pathogenesis and progression of nonalcoholic fatty liver disease by inhibiting the expression of silencing information regulator 2 related enzyme 1.

Chronic obstructive pulmonary disease (COPD) is a common, preventable and treatable disease that is characterized by persistent respiratory symptoms and airflow limitation due to airway and/or alveolar abnormalities usually caused by significant exposure to noxious particles or gases. In addition to smoking COPD caused by cigarette smoke, the phenotype of non-smoking COPD accounts for at least 1/4. Non-smoking COPD accounts for a large proportion, especially in developing countries. Non-smoking COPD and smoking COPD should be paid equal attention. More in-depth study on the prognosis of non-smoking COPD is needed.

Objective:To evaluate the clinical differences between smokers and non-smokers with chronic obstructive pulmonary disease (COPD).Methods:The clinical data of 5 183 COPD patients, including 3 688 smoking COPD patients and 1 495 non-smoking patients, were collected from the respiratory and critical medicine clinics of 12 Grade-A hospitals in Hunan Province and Guangxi Zhuang Autonomous Prefecture from December 2016 to December 2019. The general condition, clinical symptoms, acute exacerbation history and pulmonary function of the two groups were compared.Results:⑴ Non-smokers were more likely to be female, to be younger and had a lower educational level ( P<0.05). There was no difference of body mass index (BMI) and marriage statue between two groups ( P>0.05). ⑵ Clinical features: the incidence of chest distress in non-smoking COPD patients was higher than that in smoking COPD patients (86.8% and 81.4%, respectively), and the incidence of asthma was also higher than that of smoking COPD patients (86.0% and 83.1%, respectively), with statistically significant difference ( P<0.05). There was no significant difference in the incidence of cough and expectoration between the two groups ( P>0.05). The COPD assessment test (CAT) score of non-smoking COPD patients was significantly higher than that of smoking group (16.2±6.4) and (15.7± 6.5) ( P<0.05). There was no significant difference in the score of dyspnea modified medical research council dyspnea scale (mMRC) and the risk of acute exacerbation between the two groups ( P>0.05). The first second exertional volume as a percentage of predicted value (FEV 1%) and forced vital capacity (FVC) of non-smoking COPD patients were higher than those of smoking COPD patients ( P<0.05). Conclusions:Compared to smokers with COPD, non-smokers more are more likely to be female and have more severe clinical symptoms.

Objective:To explore the clinical characteristics of pediatric glucose transporter type 1 deficiency syndrome (GLUT1 DS), evaluate the efficacy and safety of ketogenic diet therapy (KDT).Methods:Clinical data of 19 children with GLUT1 DS admitted to Children′s Hospital of Fudan University, Tianjin Children′s Hospital, Shenzhen Children′s Hospital, Children′s Hospital of Nanjing Medical University and Jiangxi Provincial Children′s Hospital between 2015 and 2019 were collected retrospectively. The first onset symptom, main clinical manifestations, cerebrospinal fluid features and genetic testing results of patients were summarized, the efficacy and safety of ketogenic diet treatment were analyzed. Results:Among the 19 cases, 13 were males and 6 females. The age of onset was 11.0 (1.5-45.0) months,the age of diagnosis was 54.0 (2.8-132.0) months. Epilepsy was the first onset symptom of 13 cases. Different forms of tonic-clonic seizures were the most common types of epilepsy (7 cases with generalized tonic-clonic seizures, 5 cases with focal tonic or clonic seizures, 4 cases with generalized tonic seizures). Antiepileptic drugs were effective in 4 cases. Paroxysmal motor dysfunction was present in 12 cases and ataxia was the most common one. All patients had different degrees of psychomotor retardation. Among 17 patients received cerebrospinal fluid examination, cerebrospinal fluid (CSF) glucose level was lower than 2.2 mmol/L and CSF glucose/glycemic index was<0.45 in 16 cases, only 1 case presented normal CSF glucose level (2.3 mmol/L) and normal CSF glucose/glycemic index(0.47). SLC2A1 gene mutations were found in 16 patients, missense, frameshift and nonsense mutations were the common types with 5 cases, 5 cases and 3 cases respectively. All 19 patients were treated with ketogenic diet, which was effective in 18 cases in seizure control, 11 cases in dyskinesia improvement and 18 cases in cognitive function improvement. No serious side effects were reported in any stage of KDT.Conclusions:The diagnosis of GLUT1 DS is often late. It is necessary to improve the early recognition of the disease and perform CSF glucose detection and genetic testing as early as possible. The KDT is an effective and safe treatment for GLUT1 DS, but a small number of patients have not response to diet therapy.

Objective The aim of this study was to investigate the mechanism underlying pruritus by comparing the epidermal growth factor receptor inhibitor(EGFRI)-erlotinib mouse model with the substance P(SP)-induced pruritus mouse model. Methods Two randomized groups of mice were treated with erlotinib or SP to induce pruritus. Behavioral and skin manifestations were observed. Pathological images and neurokinin 1 receptor(NK-1R)expression of the skin were determined. Concentration of interleukin(IL)-31, IL-33, histamine, leukotriene B4, and SP was analyzed by enzyme-linked immunosorbent assay. Nitric oxide was analyzed by colorimetry. Results Transient pruritus induced by erlotinib appeared 2 to 5 days after treatment. In contrast, continuous pruritus was observed during the first hour, but was then gradually relieved. These two shared similar scratching behavior. Concentration of neurotransmitters showed similar trends in changes among the erlotinib group and SP group. Immunohistochemical expression was also consistent between the erlotinib group and SP group. Conclusions Erlotinib-associated pruritus is related to release of signaling factors through the SP/NK-1R signaling pathway.

Objective To investigate the effects of sphingosine-1-phosphate receptor 1 (S1PR1) changes on the proliferation of endogenous neural stem cells (NSCs) in hippocampus after traumatic brain injury (TBI).Methods Rat TBI models were constructed by the means of controlled cortical injury.A total of 72 rats were included and randomly divided into four groups:sham,TBI,TBI + SEW (TBI + S1PR1 agonist SEW2871 intervention) and TBI + VPC group (TBI + S1PR1 antagonist VPC23019 intervention),with 18 rats per group.The TBI model was induced by a control cortical injury device.The injured rats in TBI + SEW group and TBI + VPC group were respectively administrated with S1PR1 agonist SEW2871 and antagonist VPC23019 at scheduled time points after TBI.Hippocampal S1PR1 expression was detected by Western-blotting and the proliferation of NSCs was assessed by double-labeled immunofluorescence staining at days 7,14 and 21 after injury.Results At days 7,14 and 21 after TBI,the hippocampal S1PR1 levels and NSCs proliferation amounts in sham,TBI,TBI + SEW and TBI + VPC groups were evidently different (P ＜ 0.05).In particular,the outstanding changes among the four groups above occurred at 7 d after injury were as following:S1PR1 expression in TBI group significantly increased by 1.56 times compared with that in sham group,and it was respectively upregulated by 66.67％ in TBI + SEW group and down-regulated by 20.29％ in TBI + VPC group (P ＜0.05).The nmmber of NSCs proliferation in TBI group was 2.08 times more than that in sham group,and it increased by 36.75％ in TBI + SEW group and reduced by 18.77％ in TBI + VPC group (P ＜ 0.05).Conclusion The expression of S1 PRI is closely associated with the proliferation of NSCs in hippocampus after TBI,indicating that S1PR1 activation may be an effective strategy to improve the posttraumatic neurogenesis.

Objective To evaluate the efficacy of different concentrations and small dose of lidocaine of combined spinal epi-dural anesthesia(CSEA) in elderly patients with non-procedure for prolapse and haemorrhoids(PPH) anorectal surgery .Methods One hundred and twenty elderly patients undergoing non-PPH anorectal surgery were selected and randomly dividedin to 4 groups , the group A(1 .5% lidocaine) ,B(1 .0% lidocaine) ,C(0 .8% lidocaine) and D ,30cases in each group .The group A ,B and C adopted CSEA ,and the group D adopted the sacral anesthesia .The changes of mean arterial pressure(MAP) and heart rate(HR) at different time points ,stress reactions before and after operation ,anesthetic operating time ,effect onset time ,postoperative reactivating time , anesthetic grade ,success rate ,postoperative patient satisfaction ,postoperative surgeon satisfaction and urine retention were recor-ded .Results MAP and HR at different time points had no statistical difference among 4 groups(P>0 .05) .The anesthetic onset time and postoperative activity recovery time in the group A ,B and C were significantly less than those in the group D(P<0 .01) . The postoperative activity recovery time in the group A was longer than that in the group B and C .The urine retention time in the group A ,B and C was significantly less than that in the group D (P<0 .05) .The postoperative patient satisfaction and surgeon sat-isfaction in the group A and B were 100% .Conclusion 1% lidocaine CSEA is more suitable for the elderly patients with non-PPH anorectal operation .

BACKGROUND:Sufentanil exerts protective effects on tissues, but its roles in the repair of nervous system injury and the underlying mechanism are stil unknown.
OBJECTIVE:To explore the protective effect of sufentanil preconditioning in the repair of spinal cord injuries and the underlying mechanism.
METHODS:Mouse models of spinal cord injuries were prepared through clipping spinal cord fol owed by intraperitoneal injection of 3 and 6μg/kg sufentanil, respectively.
RESULTS AND CONCLUISON:(1) Western blotting, ELISA and TUNEL assays showed that 6μg/kg sufentanil significantly down-regulated the protein expression levels of TLR4, nuclear factor-κBp65, cleaved-caspase 3, tumor necrosis factor-αand interlenkin-1βin the spinal cord of mice (P<0.05);at the same time, the number of apoptotic neurons was significantly decreased (P<0.05). (2) Furthermore, high-dose sufentanil preconditioning significantly ameliorated the recovery of limb function at 14 days after injury (P<0.05). (3) These results administrate that the neuroprotection provided by 6μg/kg sufentanil preconditioning for spinal cord injuries in mice maybe related to the TLR4/nuclear factor-κB signaling pathway inactivation.

Objective To investigate the effect and mechanism of AngⅡ on collagen in hepatic stellate cell. Methods HSCs were isolated and cultured, 3H-pro incorporation method was used to evaluate the effects of different doses of AngⅡ on the proline syntheses. RT-PCR assay were used to assess changes in mRNA expression levels of type Ⅰ and Ⅲ procollagen. PDGFR-β mRNA and protein were determined by in situ hybridization and immunocytochemistry. Results 10-8~ 10-5 mol/L AngⅡ could significantly increase the 3H-pro incorporation rate of HSC in a dose-dependent style, 10-6 mol/L AngⅡis the most effective dose. The cultured HSC showed a little expression of type Ⅰ and Ⅲ procollagen mRNAs, while 10-6 mol/L AngⅡwas able to enhance the expression for type Ⅰ and Ⅲ procollagen mRNAs significantly(P < 0.01). AngⅡalso could enhance both mRNA and protein expression of PDGFR-β on HSC(P < 0.01). Conclusion These results suggest that AngⅡ could promote HSC collagen synthesis by enhancing the expressions of PDGFR-β.