Objective: To clone and analyze the full-length cDNA of chalcone isomerase (CHI) gene from Fagopyrum dibotrys (FdCHI). To analyze the expression of CHI and the total flavonoids content during florescence of F. dibotrys. Methods: The cDNA sequence of CHI was cloned by homology cloning from F. dibotrys. The expression of CHI was analyzed in the different tissues during florescence by semi-quantitative RT-PCR. The content of total flavonoids was measured by AlCl3 method. Results: The open reading frame (ORF) of FdCHI was 750 bp and encoded a protein with 249 amino acids. Bioinforamtion analysis suggested that the amino acid sequence of FdCHI had the high homology with those in other plants. Gene expression analysis showed that FdCHI was highly expressed in the flowers, followed by the roots and leaves, while lower in the stems. The content of total flavonoids was the highest in the flowers then the leaves and stems, and the lowest was in the roots. Conclusion: The cDNA sequence of FdCHI is firstly obtained from F. dibotrys and its coding protein has the typical characteristic of CHI homologous protein. The gene expression of FdCHI shows the same to the total flavonoids content in the stems, leaves, and flowers, but different in the roots of F. dibotrys.

OBJECTIVEThe device of a bone age film of analysis and process was designed, can calculate the expected patient's height and identify growth level and development.

METHODSSelect the DR bone age of 100 children of 6-13 years old, used semi Lagrange algorithm of target region of interest on bone age piece (ROI) for image analysis, compared with 2 pediatric endocrinologists (A, B) who used TW3 artificial to judge bone age (two methods were detected 2 times), and report the results.

RESULTSBone age assessment process, forecast error of bone age reduced to 0.12 years.

CONCLUSIONSThis device can quickly calculate the expected patient's height and identify his growth level, improve the speed and accuracy of bone age assessment, especially in the poor medical conditions in rural and remote areas.

Objective: To explore the effect of suberoylanilide hydroxamic acid (SAHA) improving cardiac hypertrophy via inhibiting histone deacetylases (HDAC) in experimental mice and to provide a new idea for prevention and treatment of cardiac hypertrophy. Methods: Cardiac hypertrophy mice model was established by thoracic aorta ligation. A total of 60 Kunming mice were randomly divided into 4 groups: Normal control group, Sham operation group, Cardiac hypertrophy (CH) group and CH+SAHA group. There were 6 mice used in each group. Myocardial cell morphology was observed by HE staining, cardiac function was assessed by echocardiography, mRNA and protein expressions of HDAC5 (the isoform of HDAC) and β-MHC were examined by RT-PCR and Western blot analysis. Results: The mice in CH group had myocardial cell hypertrophy, disordered arrangement and hyperchromatic nucleus. Compared with Sham operation group, CH group showed decreased left ventricular end diastolic diameter (LVEDD), left ventricular end diastolic volume (LVEDV) and increased thickness of inter-ventricular septum (IVS), allP<0.05; CH group presented elevated mRNA and protein expressions of HDAC5 and β-MHC,P<0.05. SAHA obviously decreased HDAC5 expression, down regulated cardiac hypertrophy related β-MHC gene expression, improved cardiac function and hypertrophy, all P<0.05. Conclusion: HDAC were involved in myocardial hypertrophy; SAHA could inhibit HDAC expression and therefore,improved myocardial hypertrophy in experimental mice.

Objective:To evaluate the effects of endovascular embolization and microsurgical clipping on cognitive function in patients with intracranial aneurysm.Methods:A total of 80 patients with intracranial aneurysms,who were treated in Chongqing Hospital of Traditional Chinese Medicine from March 2014 to September 2015,were selected and randomly divided into control group(n=40) and experimental group (n=40).The control group was treated with microsurgical clipping,and the experimental group,with endovascular embolization.The postoperative cognitive function was assessed by the mini mental state examination scale (MMSE),and length of stay,in-hospital mortality rate,mortality rate in 1 year and the incidence of postoperative adverse reactions were compared between the two groups.Results:The total score (26.78± 0.85) of MMSE in the experimental group was higher than that (22.25± 0.63) of the control group (P＜0.05).The cognitive dysfunction rate (37.50％) of the experimental group was lower than that (55.00％) of the control group (P＜0.05).The length of stay [(6.7± 3.9) days] of the experimental group was lower than that [(9.6± 4.5) days] of the control group(P＜0.05).There were no significant differences in in-hospital mortality rate and mortality rate in 1 year,and the incidence of adverse reactions between the two groups (P＞0.05).Conclusion:Endovascular interventional therapy can improve the cognitive function of the patients with intracranial aneurysms and shorten the length of hospital stay.

Cerebral apoplexy is one of the main causes of death in the middle-aged and elderly population,which has higher mortality and disability rate.The incidence of the disease is increasing year by year and it is a serious threat to human life and health.Therefore,it is of great significance to find an effective target for the diagnosis and treatment of stroke.Thioredoxin (Trx) is the major thiol reducing agent in the cells,it is involved in many signal transduction pathways in the cells by regulating the redox state of the cell.It has disulphide reductase activity,which can reduce the oxidative stress injury in the rats after the stroke.Thioredoxin interacting protein (TXNIP) is an endogenous inhibitor of Trx,it can destroy the redox balance and promote the oxidative stress by binding/inhibiting the activity of Trx,while the inhibition or knockdown of TXNIP has obvious neuroprotective effects.Recent studies suggest that Trx/TXNIP may be involved in the pathophysiology of cerebral apoplexy by a variety of pathways.This article analyses the research status of Trx/TXNIP and studies the localization of Trx system in the central nervous system and the progress of Trx system in ischemic cerebral apoplexy.It reviews the mechanism of Trx/TXNIP in cerebral apoplexy and prospectes the signaling pathways involved in the pathophysiological process of Trx/TXNIP to provide new ideas for the treatment of cerebral apoplexy.

Objective To explore the risk factors on impact of early dislocation after hip arthroplasty,and provide patients with individ-ualized treatment programs. Methods We retrospectively analyzed the clinical data of displaced femoral neck fractures in 247 patients who accepted hip arthroplasty from January 2006 to December 2011,and a total of 8 potential factors associated with occurrence of early dislocation were evaluated by univariate analysis and multivariate logistic regression analysis. Results The univariate analysis showed that sex,surgical approach and femoral head size were correlated with early dislocation (P<0. 05). Multivariate logistic regression analysis showed that female (OR=1. 862),posterolateral surgical approach (OR=3. 478),the use of 22 mm femoral heads (OR=2. 558)were independent risk factors (P<0. 05). And use of≥30 mm femoral heads(OR=0. 635) was a protective factor(P<0. 05). Conclusion Occurrence of early dislo-cation after hip arthroplasty is mainly correlated with many factors,various risk factors should be taken into account in preoperative and intrao-perative and choose the optimal individualized treatment plan.

Nicotinic acetylcholine receptors (nAChRs) belong to ligand-gated ion channel receptors, of which α7 nAChR subtype is widely distributed in the cerebral cortex, thalamus, hippocampus, and also identified in microglia, macrophages, bone marrow cells, etc. Previous studies revealed that α7 nAChR is closely related to the function of the cholinergic anti-inflammatory pathway, and is a vital target for drug development of Alzheimer's disease and schizophrenia. The establishment of a stable α7 nAChR in vitro drug screening system is crucial for the efficient screening of novel drugs targeting this target. Recombinant expression of different subtypes of nAChRs on Xenopus laevis oocyte membranes and current detected by two-electrode voltage clamp (TEVC) is an advanced and complex model for novel drug screening. Molecular chaperones can assist the assembly of some nAChR subunits to form functional receptors, providing a stable expression model for the screening of compounds targeting this receptor. In this study, a molecular chaperone gene of α7 nAChR, transmembrane protein 35A (Tmem35a), was isolated and cloned from rats. We constructed the recombinant expression vector and obtained the cRNA of Tmem35a by in vitro transcription technique. Two cRNAs (Tmem35a and α7) were mixed and injected into X. laevis oocytes for expression. Then, the effects of this molecular chaperone on the current expression and pharmacological properties of α7 nAChR were evaluated by the TEVC. The results revealed that TMEM35A, also known as novel acetylcholine receptor chaperone (NACHO) could effectively increase the expression of α7 nAChR protein on oocyte membranes, and the amount of α7 nAChR protein was increased about 1-fold. The peak current induced by agonist acetylcholine (ACh) was increased about 10-fold. After injection of Tmem35a cRNA, the median effect concentration (EC₅₀) value of α7 nAChR to agonist ACh is 228.5 μmol·L^-1, which shows almost no difference from native α7 nAChR (EC₅₀: 223.3 μmol·L^-1), indicating the preservation of the normal properties of α7 nAChR. The results of this investigation indicate that the molecular chaperone NACHO effectively assists the heterologous expression of α7 nAChR in X. laevis oocytes, which provides a model for screening the potency of lead compounds targeting α7 nAChR. All animal experiments in this study were reviewed and approved by the Ethics Committee of Guangxi University (approval number: GXU-2023-0249).

N-type voltage-gated calcium (Ca²⁺) channels (N-type VGCC, Ca_V2.2) mediate Ca²⁺ influx in response to action potential at the presynaptic terminal, and play an important role in synaptogenesis, neurotransmitter release and nociceptive signal transduction. It is a new target for the development of drugs for the treatment of neuralgia (chronic pain) and other major diseases. Due to the difficulty of calcium channel expression in vitro and the detection of channel current, there is a great lack of new drug screening models. In this study, we established and optimized the electrophysiological drug screening model using Xenopus laevis oocytes for the recombinant expression of Ca_V2.2 in vitro (this study were reviewed and approved by the Ethics Committee of Guangxi University, approval number: GXU-2023-0249). Firstly, the linear plasmids encoding cDNA of major subunit α1B and auxiliary subunits α2δ1 and β3 of rat Ca_V2.2 were used as templates for in vitro transcription to generate their related mRNA (cRNA), after which three kinds of cRNA were injected into Xenopus laevis oocytes at the mass ratio of 2∶1∶1 for expression. The two-electrode voltage clamp (TEVC) technique was used to detect the inward current produced by Ca_V2.2. At the same time, the expression conditions of Ca_V2.2 were optimized, and its gating function was characterized from the aspects of channel activation and inactivation. The results showed that 3-5 days after cRNA microinjection, stable Ca_V2.2-mediated barium ion (Ba²⁺) currents were successfully detected. The interference of endogenous potassium channels and Ca²⁺-activated chloride channels can be eliminated by tetraethylammonium hydroxide (TEAOH) and 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid tetrakis (BAPTA-AM) treatment. The maximum potential for Ca_V2.2 activation is 0 mV, and the current reverses to be outward when the membrane potential is greater than +50 mV. By fitting the steady-state activation and inactivation curves, the half-maximal activation potential and half-maximal inactivation potential of Ca_V2.2 are identified as -15.9 and -60.2 mV. In this study, a stable Ca_V2.2 expression system was established based on Xenopus laevis oocytes. The in vitro expression system can provide a new way for the screening of Ca_V2.2 active compounds or lead drugs.

Objective To comment the severity of severe hand,foot and mouth disease(HFMD)by pediatric risk of mortality score(PRISM),and assess the performance of PRISM in predicting mortality or complication probability in HFMD.Methods Four hundred and twenty-four severe HFMD pediatric patients were recruited in the study from 1th Jan 2010 to 31th June 2013.Information on the outcome and the varia-bles required to calculate PRISM score were collected.The logistic regression model developed in the learning sample was evaluated in the test sample by calculating the area under the receiver operating characteristic (ROC)curve to assess discrimination pneumorrhagia and death.Calibration across deciles of risk was evalua-ted using the Hosmer-Lemeshow goodness-of-fit χ2 test.Results The area under the ROC curve were 0.87 (95%CI 0.80~0.94 )for PRISM in predicting pneumorrhagia probability.The area under the ROC curve were 0.87(95%CI 0.80~0.95)for PRISM in predicting mortality probability.The PRISM in observed and expected pneumorrhagia did not demonstrate good calibration at ten mortality risk intervals (χ2 =36.66, P<0.001 ).The PRISM in observed and expected mortality did not demonstrate good calibration at ten mortali-ty risk intervals(χ2 =41.11,P<0.001).Conclusion The PRISM score is demonstrated good discrimination of pneumorrhagia and death in HFMD pediatric patients,but the performance of calibration is not good.

Objective To investigate the clinical spectrum of respiratory viruses in infants and young children with acute lower respiratory infection(ALRI) in Chongqing area from 2003-2007.And to assess the clinical diagnostic value of virus detection in nasopharyngeal secretions(NPS) and serum viral antibody detection for ALRI.Methods Cases of 2 529 specimens of NPS in hospitalized children with ALRI from Apr.2003 to Oct.2007 were taken for detecting 7 common respiratory virus antigens by immunofluorescence assay including respiratory syncytial virus (RSV),adenovirus(ADV),influenza A(IA),influenza B (IB),parainfluenza virus1-3 (PIV1,PIV2,PIV3).Fifty-five thousand eight hundred and eighty-seven samples were tested for ADV-IgM by ELISA.Among those,45 159 cases were further tested for RSV-IgM by ELISA.Results Respiratory virus pathogens were detected in 778 samples out of 2 529(30.76%) including RSV positive in 668 samples (85.86%),PIV3 positive in 75 samples (9.64%),IA positive in 22 samples (2.57%),ADV positive in 15 samples ( 1.93%),only 1 sample ( 0.13%) positive for both PIV1 and RSV. And the positive rate of RSV-IgM was 0.9%-15.2%,and the positive rate for ADV-IgM was about 0.6%-10.6%.RSV infection occured mainly in winter and spring.Conclusions Respiratory virus is the most common pathogen in children with ALRI during the survey period in Chongqing area,especially for RSV infection.The pattern of RSV circulation varied every year with seasonality.It is suggest that this year is peak one for RSV infection from the monthly positive results,especially in Feburary(50%) in 2007.But the infection rate of PIV3,IA,ADV and PIV1 are lower,particularly IB and PIV2 infection have not been seen for the last 5 years.It is fast and accurate to detect RSV antigen and suit to clinical diagnosis by using immunofluorescence assay than other antibody detection.