Objective To express soluble single chain variable fragments (ScFv) of monoclonal antibody MC3 recognizing colorectal carcinomas in E. coli HB2151 and to purify the soluble ScFv and identify its antigen binding activities to find new target vectors for the diagnosis and therapy of colorectal carcinomas. Methods The phage clones displaying ScFv fragment of the monoclonal antibody MC3 were used to infect E. coli HB2151 to express soluble antibodies. The soluble ScFvs were identified by Dot blot and Western blot and their antigen binding activities were determined by ELISA. The VH and VL DNAs of the ScFv DNA derived were sequenced based on the dideoxy method. Results The soluble MC3 ScFvs were expressed successfully. The expression products with a proximate MW of 32?10 3 were mainly secreted into the periplasm. The soluble ScFv containing periplasmatic extracts derived from three clones could inhibit the binding of MC3 with its antigen, and the inhibition rates were 41.19%, 36.89% and 33.77% respectively. The sequences of the VH and VL DNAs of the MC3 ScFv showed that the variable antibody genes belonged to the IgG1 subgroup and ? type. Conclusion Generation of E. coli HB2151 expressed ScFv of monoclonal antibody MC3 paves the way for further use of the antibody.

Objective To improve the diagnosis and treatment of testicular masses associated with congenital adrenal hyperplasia(CAH).Methods A 25-year-old man presented with bilateral testicular nodules.Physical examination revealed palpable multiple nodules in the bilateral testes and epididymides as well as enlarged and hard right testis with uneven surface.Biochemical examination showed testosterone(T) of 18.7 nmol/L,17?-(OH)P＞20.0 ng/ml,positive results of rapid ACTH stimulation test and dexametha- sone suppression test.B-ultrasound demonstrated diffuse testicular lesions with nodules in parts of the bilater- al testes.Bilateral testicular nodules were biopsied,and pathological diagnosis was interstitial cell hyperplasi- a.Positive staining for?-inhibin was observed immunohistochemically.The clinical characteristics of testicular masses associated with CAH were reviewed in combination with the literature.Results Based on medical history,imaging,laboratory and pathologic examinations,the patient was diagnosed with testicular masses as- sociated with CAH.He received oral cortisol at dose of 40 mg daily for 2 weeks,and then at dose of 30 mg daily as maintenance therapy.After treatment of 8 months,B-ultrasound showed complete regression of the testicular tumors.Follow-up of 24 months showed no recurrence of the tumor.Conclusions For patient with bilateral testicular masses,endocrinological evaluation and clinical history taking are indicated to exclude testicular interstitial cell hyperplasia associated with CAH.By oral steroid replacement therapy,hormone-sen- sitive testicular masses can regress.

Objective To evaluate the clinical value of 6 h 99mTc-diethyl iminodiacetic acid (99mTc-EHIDA) planar hepatobiliary scintigraphy (HBS),6 h tomographic HBS and 24 h planar HBS in diagnosis on biliary atresia(BA).Methods Seventy cases(32 male,38 female) with continuous jaundice received planar and tomographic HBS in Beijing Friendship Hospital from Jan.2005 to Dec.2007.The mean age was 48.7 d (29 d-4 months).According to final diagnosis,all cases were divided into BA group (45 cases) and non-BA group (25 cases).All cases fasted at least 4 hours before HBS.The equipment was 3 head IRIX from Philips company with low energy high resolution collimator.The tracer was 99mTc-EHIDA and the radiochemistry purity was more than 95 percent.The dosage was 7.4 MBq/kg.All diagnosis demonstrated by operation pathology and clinical follow-up.All cases received HBS at 5,10,15,20,30 min and 1,6 h after tracer injection.HBS would ended if radioactivity appeared in gallbladder or intestine.These cases would received tomographic HBS and 24 h HBS if radioactivity did not appear in gallbladder or intestine at 6 h post injection.All these images were analyzed by 2 or more nuclear medicine physicians.Results There were not radioactivity appearing in gallbladder and intestine on planar and tomographic HBS of 27 cases,which suggested the BA.There were radioactivity appearing in gallbladder and intestine on planar and tomographic HBS of 30 cases,which suggested the non-BA.Positive rate of 6 h tomographic HBS was significantly higher than that of 6 h planar HBS and there was significantly difference between the 2 methods.Positive rate of 6 h tomographic HBS was significantly higher than that of 24 h planar HBS and there was significant difference between the 2 methods.Conclusions 99mTc-EHIDA HBS is a noninvasive,safety,valuable examing method and has definitely clinical value in the diagnosis on BA.The clinical value of 6 h tomographic HBS is significantly higher than that of 6 h planar HBS and 24 h planar HBS.

Objective To estimate the difference of PS、CBV/CBF in the evaluation pf angiogenesis and growth behavior of the C6 glioma.Methods Sixty adult Wistar rats were divided into 3 groups randomly.CT perfusion were performed at the time of 5,13,20 d after the rats were inoculated C6 glioma cells.Permeability surface(PS),cerebral blood volume(CBV),cerebral blood flow(CBF)of different part of the tumor(central part,peripheral part,adjacent part and contralateral normal parenchyma)were measured at different time.Results At the central parts of the lesions,there were obvious difference between different time of tumor growth among PS[(3.94?0.15),(8.47?0.34),(5.20?0.65)ml? 100g~(-1)?min~(-1)],CBF[(280.33?8.82),(388.33?14.00),(116.16?11.54)ml? 100g~(-1)?min~(-1)],CBV[(7.75?0.27),(12.73?0.98),(5.14?0.66)ml?100g~(-1)](F=4.421,P= 0.013;F=11.370,P=0.000;F=15.789,P=0.000).There were statistical difference of PS at the different time in both the peripheral and adjacent parts of the glioma.(F=13.567,P=0.000;F=12.470, P=0.000).No difference were detected in CBF or CBV at different time of the peripheral parts of the tumors(F=1.176,P=0.336;F=0.148,P=0.710).there were significant difference between different time of tumor growth among CBF[(175.33?12.95),(275.50?13.76),(246.33?12.81)ml? 100g~(-1)?min~(-1)],CBV[(4.15?0.47),(8.05?0.30),(7.54?0.89)ml?100g~(-1)]at the adjacent parts of the tumors(F=24.176,P=0.000;F=17.148,P=0.000;F=15.791,P=0.000). Coneluslon CBV,CBF can reflect the number and volume of the tumor vessels,while PS can directly reflect the function of the angiogenesis and the behavior of the glioma.

OBJECTIVETo demonstrate the chemical constituents of Alyxia sinensis.

METHODPhytochmical experiment was carried out, using column chromatograph technologies.

RESULTFive compounds have been isolated from the petroleum ether soluble part of the stems of A. sinensis. Their structures have been elucidated respectively as heptatriacontane(1), octatriacontane(2), 20-noatriacontannone(3), 20-nonatriacontanone(4), 20-tetraacontanoe(5), physcion(6), emodin(7), chrysophanol(8), coumarin(9), stigmasterol acetate(10), beta-sitosterol acetate(11), lupeol(12), betulin(13), stigmasterol(14), beta-sitosterol(15), ursolic acid(16), oleanolic acid(17).

CONCLUSIONAll these compounds are firstly been isolated from A. sinensis. And compound 5-9, 10, 11 are also been separated from Alyxiae genus for the first time.

Anthraquinones ; chemistry ; isolation & purification ; Apocynaceae ; chemistry ; Emodin ; analogs & derivatives ; chemistry ; isolation & purification ; Plant Stems ; chemistry ; Plants, Medicinal ; chemistry

Background Proliferation of the human Tenon capsule fibroblasts(HTFs) is a main cause of failure of filtering surgery.To search the drug of inhibiting the growth of the HTFs is essential for the improvement of successful rate of filtering surgery.Objective The aim of this study was to investigate the inhibitory effect of apigenin on HTFs and its mechanism.Methods Human Tenon capsular tissue was obtained during the strabismus correction surgery.HTFs was primarily cultured using explant method and identified using vimentin by immunochemistry.The 3-5 generation of cells were incubated to 96-well plate.Apigenin of 0,20,40,80,160 μmol/L was added into the medium,respectively,for 24,48,72 hours,and the proliferation of HTFs was detected by sulfonyl chloride (SRB) at the wavelength of 560 nm (A560).Bromodeoxyuridine (BrdU) of 10 μg/L was added to culture the cells for 48 hours to calculate the labeling rate of BrdU.The morphology of the cells was observed using Hoechst 33258 staining,and apoptosis and cells cycle were evaluated by flow cytometry.Results Cultured cells grew well with the positive response for vimentin,showing the green fluorescence in cytoplasm.SRB assay showed that the A560 value was gradually declined with the increase of the dosage of apigenin and prolong of time (Fgroup =480.306,P =0.000 ; Ftime =555.144,P =0.000).The labeling rate after 0,40,80 μmol/L apigenin acted for 48 hours was (87.860 ±0.632)％,(61.520±4.306)％ and (23.480±4.472)％,showing a significant difference among the three groups (F =299.347,P =0.000).The labeling rate of HTFs for BrdU was significantly decreased in the 40 and 80 μmol/L apigenin groups compared with the 0 μmol/L apigenin group (P＜0.05).Hoechse 33258 staining found that the number of the HTFs was gradually decreased and the cell number of karyopyknosis and nuclear deformation was increased with the increase of apigenin dosage.Percentage of cells in G0/G1 phase were raised and that in S and G2/M phase were declined in the higher dosage apigenin group,with a significant difference among the different groups (FG0/G1 =58.621,P=0.000;Fs =32.357,P=0.001 ;FG2/M =83.998,P=0.000).In the 72nd hour after acted by 0,40,80,160 μmol/L apigenin,the apoptosis rate of HTFs was (4.77±0.21) ％,(13.24±1.35)％,(18.33±1.86) ％,(31.58 ± 2.77) ％,respectively,with a statistically significant difference among the four groups (F =204.791,P＜0.05).Conclusions Apigenin restrains the growth of HTFs by evoking G0/G1 cell cycle arrest and inducing apoptosis in a dosage-and time-dependent manner.

BACKGROUNDEvaluation of the severity of the pregnant women with suitable admission to the Intensive Care Unit (ICU) is very important for obstetricians. By now there are no criteria for critically ill obstetric patients admitted to the ICU. In this article, we investigated the admission criteria of critically ill patients admitted to the ICU in order to provide a referral basis of reasonable use of the ICU.

METHODSA retrospective analysis of critically ill pregnant women admitted to the ICU in Perking University Third Hospital in China in the last 6 years (from January 2006 to December 2011) was performed, using acute physiology and chronic health evaluation II (APACHE-II), Marshall and WHO near miss criteria to assess the severity of illness of patients.

RESULTSThere were 101 critically ill pregnant patients admitted to the ICU. Among them, 25.7% women were complicated with internal or surgical diseases, and 23.8% women were patients of postpartum hemorrhage and 23.8% women were patients of pregnancy-induced hypertension. Sixty-nine cases (68.3%) were administrated with adjunct respiration with a respirator. Sixteen cases (15.8%) required 1-2 types of vasoactive drugs. Fifty-five cases (54.5%) required a hemodynamic monitoring. Seventy-three cases (72.3%) had multiple organ dysfunctions (MODS). The average duration in ICU was (7.5 ± 3.0) days. A total of 12.9%, 23.8% and 74.3% of women were diagnosed as critically ill according to the APACHE-II, Marshall and WHO near miss criteria, respectively. The rate was significantly different according to the three criteria (P < 0.01).

CONCLUSIONSThe WHO near miss criteria can correctly reflect the severity of illness of pregnant women, and the WHO near miss criteria are appropriate for admission of critically ill pregnant women to ICU in China.

APACHE ; China ; Critical Illness ; Female ; Humans ; Intensive Care Units ; Pregnancy ; Retrospective Studies ; Severity of Illness Index ; World Health Organization

Objective To investigate the pathogenesis of osteoarthritis and feasibility of acupotomy therapy through the observation on the influences of acupotomy therapy on ethology and morphology in rabbits with knee osteoarthritis.Methods The model of knee osteoarthritis was established by applying the fixed method of knee extension posture in rabbits. The therapy of electro-acupuncture and acupotomy therapy were used to treat the rabbits. After 3 weeks the knee joint function was observed and reviewed based on Lequesne algofunctional index. The light microscope was used to observe tibia tissue after HE staining. The transmission electron microscope (TEM) was used to observe the internal femoral condyle cartilage.Results Lequesne algofunctional index showed that in local reaction, gait reaction and joint motion there was a significant different between the electro-acupuncture group or acupotomy group and the normal group or model group (P<0.05), respectively, while there was no different between the electro-acupuncture group and acupotomy group (P>0.05). HE staining showed that the cartilage injury was relieved and tended to recovery in the electro-acupuncture group and acupotomy group. The result of TEM showed that the lesion of cartilage cells and disorder of collagen fibrils were relieved in the acupotomy group, which is similar to those in the electro-acupuncture group.Conclusion The acupotomy therapy has a good curative effect on knee osteoarthritis, which is proved at macroscopic and microscopic levels.

BACKGROUNDDuloxetine is approved for the treatment of generalized anxiety disorder (GAD) in the United States and elsewhere. This study aimed to assess the efficacy, tolerability, and safety of duloxetine in Chinese patients with GAD.

METHODSThis 9-site study consisted of double-blind treatment for 15 weeks either with duloxetine 60 - 120 mg or with placebo. Patients with at least moderately severe GAD and a Sheehan Disability Scale (SDS) global functioning impairment total score ≥ 12 were included in this study. Patients who were randomly assigned to duloxetine received 60 mg for 7 weeks; at that point, for nonresponders the dose was increased to 120 mg for the remaining 8 weeks. The primary efficacy measure was mean change from baseline to endpoint on the Hospital Anxiety and Depression Scale-Anxiety subscale score (HADS-A). Secondary efficacy measures included the Hamilton Anxiety Rating Scale (HAMA), the SDS, and pain measures. Safety and tolerability were assessed.

RESULTSBaseline characteristics did not differ significantly between treatment groups. Mean age of the subjects (n = 210) was 37.6 years, 50.5% were female, and 74.3% completed the 15 weeks treatment. Patients treated with duloxetine had significantly greater improvement compared to placebo on the HADS-A (mean change -6.6 vs. -4.9, respectively, P = 0.022). Improvement in anxiety was greater with duloxetine treatment at 7 weeks and continued through 15 weeks for both the HADS-A and the HAMA total score (0.01 ≤ P < 0.05). Compared with placebo, duloxetine was also associated with greater improvement on most secondary measures, but not on the SDS global functioning score. Nausea, dizziness, and somnolence occurred significantly more frequently as treatment-emergent adverse events with duloxetine treatment compared with placebo treatment.

CONCLUSIONSDuloxetine 60 - 120 mg once daily is effective and well-tolerated for the treatment of patients with GAD in China.

Adult ; Antidepressive Agents ; adverse effects ; therapeutic use ; Anxiety Disorders ; drug therapy ; Double-Blind Method ; Duloxetine Hydrochloride ; Female ; Humans ; Male ; Middle Aged ; Thiophenes ; adverse effects ; therapeutic use ; Treatment Outcome