AIM:To detect the expression of miRNA-363 and SOX4 in osteosarcoma tissues and to investigate the effect of miRNA-363 on the viability and apoptosis of human osteosarcoma cell line MG-63.METHODS: Real-time PCR was used to detect the expression level and the relationship of miRNA-363 and SOX4 mRNA in the osteosarcoma tis-sues and the corresponding paratumorous tissues collected from 63 patients.The expression levels of miRNA-363 and SOX4 in osteosarcoma cell line MG-63 after transfected with miRNA-363 mimics were measured .The cell viability was measured by CCK-8 assay.Flow cytometry was used to monitor the changes of cell cycle and apoptosis .The changes of SOX4 and miRNA-363 expression levels in the MG-63 cells after transfection with SOX4 siRNA or pcDNA/SOX4 was detect by real-time PCR.RESULTS:The expressed level of miRNA-363 was lower , and the expression level of SOX 4 was higher in the osteosarcoma tissues than those in the adjacent normal tissues .A significantly negative correlation between the expression levels of miRNA-363 and SOX4 was observed .The expression of miRNA-363 in the MG-63 cells after transfection with miRNA-363 mimics was significantly up-regulated, while the expression of SOX4 in the MG-63 cells was significantly down-regulated , with significant difference as compared with the cells transfected with miRNA-NC and control cells .The viability of MG-63 cells was inhibited, the cell cycle was arrested in G0/G1 phase, and the cell apoptosis was increased by transfec-tion with miRNA-363 mimics.The relative protein expression levels of SOX 4 in SOX4 siRNA group and pcNDA/SOX4 group were significantly different from those in negative control group , but the relative expression levels of miRNA-363 had no significant difference .Over-expression of SOX4 restored the viability of the MG-63 cells reduced by miR-363.CON-CLUSION:The expression level of miRNA-363 is low in human osteosarcoma tissue .miRNA-363 may inhibits the viabili-ty of osteosarcoma cell line MG-63 and promotes cell apoptosis in vitro via inhibiting the SOX4 expression.

Objective To explore the clinical characteristics and therapy of brucellar meningitis in children.Method One case of brucellar meningitis was reported and relevant literatures were reviewed.A 10 years old boy was admitted to hospital with a history of half a month fever and cough and 3 days headache.His family raised sheep,but he denied contacting closely with sheep and eating raw goat's milk and mutton.Results The course of this reported child was subacute.Clinical manifestations included fever,headache and meningeal irritation sign positive.The cerebrospinal fluid(CSF)findings were similar to those of tuberculous meningitis,where WBC count was elevated with predominant lymphocytes,glucose and chloride decreased and protein increased.The serum bacterium burgeri antibody was positive,agglutination titer was 1160.After brucellar meningitis was diagnosed,three antibiotics including rifampin,minocycline and ceftriaxone sodium and other symptomatic treatment were used.The CSF findings were normal 2 weeks later.This child taked orally minocycline,rifampin and trimethoprim-sulfamethoxazole for 8 weeks post discharge,the recovery was good.The domestic and foreign documents showed that the incidence of the nervous system complications of brucellosis in children was lower than that in adult.The main clinical manifestations were acute meningitis /meningocephalitis.Brucellar meningitis could be misdiagnosed as tuberculous meningitis,because the course of disease,clinical manifestations and CSF findings were similar.Therapeutic alliance including 3 kinds of antibiotics,which could easily pass through blood-brain barrier,was used to treat brucellar meningitis.The total course of treatment shouldn't be shorter than 8 weeks.Conclusions Brucellar meningitis should be screened through epidemiological investigation,serum agglutination test and cerebrospinal fluid culture,in case the reason of meningitis of meningoencephalitis is unknown,especially its clinical manifestation resembles to tuberculous meningitis.

OBJECTIVETo explore effective therapeutic methods for sensorineural hearing loss (SNHL).

METHODSThirty-four cases of mild and moderate SNHL who were definitely diagnosed with pure tone test and by otologists, including 24 cases of insufficiency of the liver and kidney, 10 cases of excess of liver-fire. They were treated with acupuncture at main points, Ermen (TE 21) or Tinggong (SI 19) or Tinghui (GB 2), and Yifeng (TE 17), Shenting (GV 24), Baihui (GB 20), Houding (GV19), Touwei (ST 9), Conger 1-3, Congnao 1-2; and adjuvant acupoints: Huangshu (KI 16), Qihai (CV 6), Guanyuan (CV 4), Taixi (KI 3) were added for insufficiency of the liver and kidney; Zhongwan (CV 12), Tianshu (ST 25), Yanglingquan (GB 34) and Taichong (LR 3) were added for excess of liver-fire. Sequential multipoint pulse stimulation was adopted, twice each week, for 20 consecutive weeks (40 sessions) and then pure tone test was carried out repeatedly.

RESULTSOf 34 cases (58 affected ears), 13 cases were cured and 21 improved, and the various linguistic frequency hearing threshold pure hearing values before and after treatment increased significantly (P < 0.05).

CONCLUSIONAcupuncture and continuous multipoint pulse stimulation has definite therapeutic effect on linguistic frequency hearing loss in the patient of SNHL.

Acupuncture Points ; Acupuncture Therapy ; Hearing Loss, Sensorineural ; Hearing Loss, Sudden ; therapy ; Humans ; Linguistics

BACKGROUND: Schizophrenia is substantially heritable, but specific susceptibility genes remain difficult to be identified. Therefore, it is necessary to explore hereditary markers first.OBJECTIVE: To investigate the relationship between schizophrenia and related vWA allele genes based on the analysis of microsatellite DNA vWA polymorphism.DESIGN: A case-controlled study with schizophrenic patients and randomly selected population as subjects.SETTING: Ward of Dalian Seventh People's Hospital and Molecular Biological Laboratory of Dalian Medical University.between March and July 2002 at Dalian Seventh People's Hospital which specializes in schizophrenia. Schizophrenia was diagnosed according to the diagnostic standard of the third edition of "the American Diagnostic Statistical Manual for Schizophrenic Diseases", and their clinical manifestations were predominantly negative signs. Altogether 123 normal blood samples were collected from random population at the Blood Center of Dalian Red Cross. They all denied psychological ailments and severe systematic diseases, and they had no kinship with each other.METHODS: Heparin anti-coagulation blood samples were collected and PCR compound amplification was carried out with the aid of PE Profiler plus system. Then the products were subjected to electrophoresis and gene detection with ABI310 type gene analysis system so as to calculate the frequency of allele genes; Hardy-Weinberg equation law was used to make coincidence test and linkage analysis of the theoretical frequency and actual one. Schizophrenic patients and random population were compared and relative risk was calculated with RR=Pd × (1-Pc)/Pc × (1-Pd) in order to assess the statistical significance (RR: relative risk; Pd: gene frequency of schizophrenia; Pc: gene frequency of random population). RR ＞ 1 was considered of higher susceptibility while RR ＜ 1 was considered of anti-susceptibility. In this way, we could find out vWA allele genes that had susceptible linkage or anti-linkage with schizophrenic related genes.MAIN OUTCOME MEASURES: Major outcome: Correlation analysis of vWA allele genes in schizophrenic patients and random population. Secondary outcome: The coincidence of vWA allele gene frequency in patients with schizophrenia and random population with what was calculated by Hardy-Weinberg law.RESULTS: Data of the two groups were processed according to the objective and statistically analyzed.① vWA allele gene frequency in patients with schizophrenia and in random population was found to coincide with HardyWeinberg law(P ＞ 0.05).② The positive rate of vWA-14 in schizophrenic patients (17.2%) was obviously different from that in random population (33.3%) (RR=0.415, P=0.014). The positive rate of vWA-17 in schizophrenic patients (31.3%) was found to be significantly higher than that in random population (19.5%) (RR=1.866, P=0.043) while it did not differ significantly in other allele genes (P ＞ 0.05).CONCLUSION: The positive rate of vWA-14 was significantly lower in schizophrenic patients than in random population, indicating that vWA-14locus may be negatively selected in schizophrenia due to some reasons,which may be approximate to anti-schizophrenia genes. Moreover, the higher expression of vWA-17 in schizophrenic patients than in random population suggests that vWA-17 locus is correlated with schizophrenia,which may be approximate to schizophrenia-susceptibile genes.

Multimedia computing technology teaching is a new kind teaching method. It conquers many shortcomings such as poor video, poor expression in traditional teaching of microbiology. But it can also strangle the improvising creation of different teachers in teaching and then lead it to be typical " computer teaching" . So we should obey the teaching discipline in making multimedia computing technology courseware. It can lead many sections part from the class teaching if just emphasize the full use of video and audio. We should make the purpose obviously, and make it as easy as possible in teaching, and we should also explore the fixed regularity to make the teaching and studying integrated perfectly.

OBJECTIVE: To realize the automated assessment of the levels of epiphysis of distal radius and ulna by support vector machine (SVM). METHODS: The X-ray films of the left wrist joints were taken from 140 teenagers aged from 11 to 19 years old as training samples. The levels of epiphysis of distal radius and ulna were divided into five developmental levels. Each level contained 28 samples. Another 35 cases were selected as independent verifying samples. SVM classification models of the five developmental levels of epiphysis of distal radius and ulna were established. The internal cross validation was made by leave one out cross validation (LOOCV), while the external validation was made by histogram of oriented gradient (HOG), and then the accuracy (PA) of testing results was calculated, respectively. RESULTS: The PA of SVM, LOOCV and HOG of distal radius epiphyseal level were 100%, 78.6%, and 82.8%, respectively; whereas the PA of SVM, LOOCV and HOG of distal ulna epiphyseal level were 100.0%, 80.0% and 88.6%, respectively. CONCLUSION The SVM-based automatic models of the growth stage of distal ra- dius and ulna appear to have certain feasibility, and may provide a foundation for software development of bone age assessment by forensic medicine.
Adolescent ; Bone Development/physiology* ; Child ; Epiphyses/growth & development* ; Female ; Humans ; Image Processing, Computer-Assisted/methods* ; Male ; Radius/growth & development* ; Support Vector Machine ; Ulna/growth & development* ; Wrist/growth & development* ; Wrist Joint/growth & development* ; Young Adult

OBJECTIVETo observe the intervention effect of Leihong Granule (LG) in in-stent restenosis (ISR) after endovascular therapy for lower extremity arterial occlusive diseases (LEAOD).

METHODSRecruited 80 LEAOD patients who successfully underwent endovascular therapy (balloon dilation and stent implantation) were randomly assigned to two groups, the control group and the LG group, 40 in each group. Patients in the control group received basic treatment, while those in the LG group additionally took LG for 3 months. Plasma levels of IL-10, IL-18, CRP, and the intima-media thickness (IMT) of lower extremity artery were observed in the two groups between and after treatment. The rate of stent patency, ABI, intermittent claudication, rest pain, and the incidence of amputation the two groups were recorded and observed in the two groups.

RESULTSIn the control group, serum levels of IL-10, IL-18, CRP, and IMT were significantly higher one month after surgery than before surgery (P < 0.05). There was no significant difference in serum levels of IL-10, IL-18, CRP, or IMT between the two groups before surgery (P > 0.05). These indices were obviously lower in the LG group than in the control group after surgery (P < 0.05). Compared with the control group, the incidence rates of intermittent claudication and the rest pain at 6 months and 12 months after surgery significantly decreased (P < 0.05). The stent patency rate at 6 months and 12 months after surgery, and ABI were significantly higher than those of the control group (P < 0.05). There was no statistical difference in the amputation rate between the two groups (P > 0.05).

CONCLUSIONLG might effectively improve ischemic symptoms of affected limbs possibly through lowering the ISR rate after endovascular therapy for LEAOD through preventing immunosuppressive actions.

Aged ; Aged, 80 and over ; Arterial Occlusive Diseases ; therapy ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Graft Occlusion, Vascular ; therapy ; Humans ; Interleukin-10 ; blood ; Interleukin-18 ; blood ; Lower Extremity ; blood supply ; Male ; Middle Aged ; Phytotherapy ; Stents ; Treatment Outcome

The chemical constituents of 95% ethanol extract of Melastoma dodecandrum were isolated and purified by chromatography on silica gel, Sephadex LH-20, and HPLC, to obtain thirteen compounds eventually. On the basis of their physico-chemical properties and spectroscopic data, these compounds were identified as quercetin (1), quercetin-3-O-β-D-glucopyranoside (2), quercetin-3-O-(6"-O-p-coumaroyl) -β-D-glucopyranoside (3), kaempferol (4), kaempferol-3-O-β-D-glucopyranoside (5), kaempferol-3-O- [2",6"-di-O-(E)-coumaroyl]-β-D-glucopyra-noside (6), luteolin (7), luteolin-7-O-(6"-p-coumaroyl) -β-D-glucopyranoside (8), apigenin (9), apigenin-7-(6"-acetyl-glucopyranoside) (10) , naringenin (11), isovitexin (12), and epicatechin-[8,7-e] -4β-(4-hydroxyphenyl)-3,4-dyhydroxyl-2(3H)-pyranone (13). Eight compounds(3,5,6,8-11 and 13) were obtained from M. dodecandrum for the first time.
Apigenin ; analysis ; Chromatography ; methods ; Chromatography, High Pressure Liquid ; Dextrans ; Flavanones ; analysis ; Flavonoids ; analysis ; chemistry ; Glycosides ; analysis ; chemistry ; Kaempferols ; analysis ; Luteolin ; analysis ; Magnoliopsida ; chemistry ; Plants, Medicinal ; chemistry ; Quercetin ; analysis ; Silica Gel

OBJECTIVETo explore the relationship between Chinese medicine syndrome patterns (CMSP) and their associated factors in patients with type 2 diabetic nephropathy stage III (DN2-3).

METHODSRetrospective analysis was conducted on 209 patients with type 2 diabetes mellitus (T2DM). The patients were allocated into two groups, the DN2-3 group and the control group. Some related clinical materials and laboratory indexes, including age, course of disease, body mass index (BMI), glycosylated hemoglobin A1c (HbA1c), systolic blood pressure (SBP), diastolic blood pressure (DBP), blood levels of total cholesterol (TC), triglyceride (TG), high and low density lipoprotein (HDL and LDL), serum creatinine (Scr) and microalbuminuria (MALB) as well as their CMSP (both the essential syndrome and the superficial syndrome) in the DN2-3 group were collected and compared.

RESULTSSignificant differences were found between the DN2-3 group and the control group in aspects of course of disease (months, 107.74 +/- 96.19 vs. 82.03 +/- 79.10), BMI (kg/m2, 26.25 +/- 4.02 vs. 24.95 +/- 3.56) and Scr level (mmoL/L, 71.93 +/- 24.24 vs. 65.91 +/- 13.70, P < 0.05). The qi-yin deficiency SP (38 cases, 36.19%), and the blood stasis (51 cases, 48.58%) presented as the dominant essential and superficial CM-SP respectively in DN2-3 patients, holding the highest proportion. Analysis on the relationship of associated indices among patients with different CMSP showed statistical differences presented in level of MALB, i.e. which in pi-shen qi-deficiency SP (128.77 +/- 103.59 mg/24 h) was higher than in yin-deficiency dryness-fire SP and qi-yin deficiency SP (88.43 +/- 68.93 mg/24h and 82.60 +/- 55.22 mg/24 h, P < 0.05); it also presented in HbA1c (%) and TG levels(mmol/L), those in stasis SP were 10.73 +/- 2.71 and 2.29 +/- 1.58 ), in dampness SP were 8.80 +/- 2.19% and 4.37 +/- 5.92, and in stasis-phlegm SP were 8.83 +/- 2.09 and 2.40 +/- 2.18 (all P < 0.05).

CONCLUSIONSThe risk factors for occurrence of DN2-3 may be the course of disease, BMI and Scr. Qi-yin deficiency with blood-stasis is the most commonly encountered syndrome in patients with DN2-3. Relations of MALB with Pi-Shen qi-deficiency pattern; HbA1c with blood-stasis pattern, and TG with dampness syndrome are distinctly exhibited in them.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Albuminuria ; physiopathology ; Diabetes Mellitus, Type 2 ; physiopathology ; Diabetic Nephropathies ; physiopathology ; Diagnosis, Differential ; Female ; Humans ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Retrospective Studies ; Young Adult

We revisited the "Xiao Chaihu Decoction event (XCHDE)" occurred in late 1980s in Japan and the Bupleuri Radix related adverse drug reaction (ADR) reports in China After careful review, comparison, analysis and evaluation, we think the interstitial pneumonitis, drug induced Liver injury (DILI) and other severe adverse drug envents (ADEs) including death happened in Japan is probably results from multiple factors, including combinatory use of XCHDE with interferon, Kampo usage under modern medicine theory guidance, and use of XCHD on the basis of disease diagnosis instead of traditional Chinese syndrome complex differentiation. There are less ADE case reports related to XCHD preparation in China compared to Japan, mostly manifest with hypersensitivity responses of skin and perfuse perspiration. The symptoms of Radix Bupleuri injection related ADEs mainly manifest hypersensitivity-like response, 2 cases of intravenous infusion instead of intramuscular injection developed hypokalemia and renal failure. One case died from severe hypersensitivity shock. In Chinese literatures, there is no report of the interstitial pneumonitis and DILI associated with XCHDG in Japan. So far, there is no voluntary monitoring data and large sample clinical research data available. The author elaborated the classification of "reevaluation" and clarified "re-evaluation upon events" included the reaction to the suspected safety and efficacy events. Based on the current status of the clinical research on the Radix Bupleuri preparations, the author points out that post-marketing "re-evaluation upon suspected event" is not only a necessity of continuous evaluation of the safety, efficacy of drugs, it is also a necessity for providing objective clinical research data to share with the international and domestic drug administrations in the risk-benefit evaluation. It is also the unavoidable pathway to culture and push the excellent species and famous brands of TCM to the international market, in that it enhances the pharmaceutical manufactures the ability to control enterprise risk and eliminate the negative impact on traditional Chinese medicine. With the widely used and ADR report concentrated Radix Bupleuri injection as an example, the author provide a new approach to post-marketing clinical research.
Bupleurum ; chemistry ; Drug Compounding ; Drug-Related Side Effects and Adverse Reactions ; etiology ; Drugs, Chinese Herbal ; adverse effects ; Humans ; Product Surveillance, Postmarketing ; methods ; Safety