Objective:To investigate the modulating effects of anti-epidermal growth factor monoclonal antibody Cetux- imab(C225)on the ehemosensitivity and radiosensitivity in a Docetaxel-resistant human lung adenocarcinoma cell line SPC-A-1/doeetaxel.Methods:Radiosensitivity of SPC-A-1/docetaxel was determined by clone formation experiment and quantified by calculating the enhancement ratio(ER).The growth inhibition of SPC-A-1/docetaxel cell line caused by C225 or combination of C225 and Docetaxel in different orders was detected by MTT assay.The effect of C225 on cell cy- cle distribution and apoptosis was determined by flow cytometry.Results:C225 combined with radiation significantly de- creased the number of the cell clones than radiation alone;the D_0 values were 1.73 Gy for the former and 2.39 Gy for the latter,and the enhancement ratio was 1.38.C225 alone at concentration up to 1000?g/ml for 48h had neither cytotoxic nor eytostatie effect on SPC-A-1/docetaxel in vitro.C225 administration followed by Docetaxel significantly decreased the ICw of Docetaxel(85.2?g/ml vs 128.7?g/ml).Flow cytometry demonstrated that C225 exposure induced apoptosis of SPC-A-1/docetaxel cells in a time-dependent manner.The cell in the G_0/G_1 fraction increased from(43.80?4.46)% to (60.50?6.57)%(P

Objective: To construct reference standards for detection and quantification of Klebsiella pneumoniae (K. pneumoniae) with SYBR Green I-based real-time PCR assay. Methods: Primers were designed based on the published sequence of the phoE gene of K. pneumoniae. The standard was prepared by cell culture, PCR and T-A clone methods, and was identified by colony PCR and DMA sequencing. Results: The standard curve showed a very good linear negative regression between threshold cycle (Ct) and Log starting quantity of copy number. The detection range was from 5.2 to 5.2 × 106 copies per reaction, and the detection limit was 6 copies per reaction. The coefficients of variance (CVs) of three parallel experiments were in the range of 0.05 %-0.91 %. Conclusion: The reference standards have high stability and reproducibility. They can be used in the quantitative detection of K. pneumoniae.

Objective: To construct reference standards for detection and quantification of Klebsiella pneumoniae (K. pneumoniae) with SYBR Green I-based real-time PCR assay. Methods: Primers were designed based on the published sequence of the phoE gene of K. pneumoniae. The standard was prepared by cell culture, PCR and T-A clone methods, and was identified by colony PCR and DMA sequencing. Results: The standard curve showed a very good linear negative regression between threshold cycle (Ct) and Log starting quantity of copy number. The detection range was from 5.2 to 5.2 × 106 copies per reaction, and the detection limit was 6 copies per reaction. The coefficients of variance (CVs) of three parallel experiments were in the range of 0.05 %-0.91 %. Conclusion: The reference standards have high stability and reproducibility. They can be used in the quantitative detection of K. pneumoniae.

Objective To compare the incidence of limb dysfunction, sensory disturbance and lymphedema after different treatment methods in breast cancer patients, and evaluate the clinically relevant factors of limb dysfunction. Methods According to the different processing operation of axillary lymph nodes, 235 patients with breast cancer during 2005 to 2012 were divided into axillary lymph node dissection group in Mastoscopy (group A, n=120) and routine axillary lymph node dissection group (group B, n=115). The flexion, extension, abduction, adduction activity of the ipsilateral shoulder were compared with the contralateral, and the circumference of the up-and-down 15 cm of both upper limbs olecroanon were measured. The incidence of limb dysfunction of the patients was followed up in 6 months, 1 year and 2 years. Results There was no significant difference in limb activity and sensory disturbance between 2 groups 6 months, 1 year and 2 years after operation (P>0.05), the incidence of limb lymphedema was higher in group B than in group A (P<0.05). Radiotherapy was related with limb lymphedema in both groups (P<0.05). Conclusion Compared with the conventional axillary lymph node dissection, endoscopic axillary lymph node dissection can reduce the incidence of limb lymphedema after operation, but there was no significant difference in the limb activity and sensory disturbance between 2 groups. Postoperative radiotherapy is a risk to increase the occurrence of limb lymphedema.

The design space of the high shear wet granulation process was established and validated within the framework of quality by design (QbD). The system of microcrystalline cellulose-de-ioned water was used in this study. The median granule size and bulk density of granules were identified as critical quality attributes. Plackeet-Burmann experimental design was used to screen these factors as follows: dry mixing time, the impeller and chopper speed of dry mixing, water amount, water addition time, wet massing time, the impeller and chopper speed of wet massing and drying time. And the optimization was implemented with the central composite experimental design based on screened critical process parameters. The design space of the high shear wet granulation process was established based on the quadratic polynomial regression model. Since the P-values of both models were less than 0.05 and values of lack of fit were more than 0.1, the relationship between critical quality attributes and critical process parameters could be well described by the two models. The reliability of design space, illustrated by overlay plot, was improved with the addition of 95% confidence interval. For those granules whose process parameters were in the design space, the granule size could be controlled within 250 to 355 μm, and the bulk density could be controlled within a range of 0.4 to 0.6 g x cm(-3). The robustness and flexibility of the high shear wet granulation process have been enhanced via the establishment of the design space based on the QbD concept.
Cellulose ; chemistry ; Reproducibility of Results ; Technology, Pharmaceutical ; methods ; Water

AIM: To observe the influence of serum contained Tongmai herbal medicine on calcium overload of the anoxic myocardial cells. METHODS: Serum contained Tongmai herbal medicine was prepared with the serum pharmacological method and the model of anoxic myocardial cells was established. The intracellular free calcium concentration and the expression of the L-type calcium channel were determined with Fura-2/AM and RT-PCR, respectively. RESULTS: The serum contained Tongmai herbal medicine and levo-anlodipine decreased the intracellular free calcium concentration in the anoxic myocardial cells. The serum contained Tongmai herbal medicine also suppressed the expression of L-type calcium channel in the anoxic myocardial cells, but levo-anlodipine had no effect on the expression of L-type calcium channel. CONCLUSION: Tongmai herbal medicine can attenuate calcium overload in anoxic myocardial cells.

Asian People/genetics* ; Gene Frequency ; Genetics, Population ; Humans ; Microsatellite Repeats/genetics* ; Phylogeny ; Polymorphism, Genetic/genetics*

Previous studies proposed that the synergistic effect of fibroblast growth factor-21 (FGF-21) and insulin may be due to the improvement of insulin sensitivity by FGF-21. However, there is no experimental evidence to support this. This study was designed to elucidate the mechanism of synergistic effect of FGF-21 and insulin in the regulation of glucose metabolism. The synergistic effect of FGF-21 and insulin on regulating glucose metabolism was demonstrated by investigating the glucose absorption rate by insulin resistance HepG2 cell model and the blood glucose chances in type 2 diabetic db/db mice after treatments with different concentrations of FGF-21 or/and insulin; The synergistic metabolism was revealed through detecting GLUT1 and GLUT4 transcription levels in the liver by real-time PCR method. The experimental results showed that FGF-21 and insulin have a synergistic effect on the regulation of glucose metabolism. The results of real-time PCR showed that the effective dose of FGF-21 could up-regulate the transcription level of GLUT1 in a dose-dependent manner, but had no effect on the transcription level of GLUT4. Insulin (4 u) alone could up-regulate the transcription level of GLUT4, yet had no effect on that of GLUT1. Ineffective dose 0.1 mg kg(-1) FGF-21 alone could not change the transcription level of GLUT1 or GLUT4. However, when the ineffective dose 0.1 mg x kg(-1) FGF-21 was used in combination with insulin (4 u) significantly increased the transcription levels of both GLUT1 and GLUT4, the transcription level of GLUT1 was similar to that treated with 5 time concentration of FGF-21 alone; the transcription level of GLUT4 is higher than that treated with insulin (4 u) alone. In summary, in the presence of FGF-21, insulin increases the sensitivity of FGF-21 through enhancing GLUT1 transcription. Vice versa, FGF-21 increases the sensitivity of insulin by stimulating GLUT4 transcription in the presence of insulin. FGF-21 and insulin exert a synergistic effect on glucose metabolism through mutual sensitization.
Animals ; Blood Glucose ; Diabetes Mellitus, Experimental ; metabolism ; Drug Synergism ; Fibroblast Growth Factors ; pharmacology ; Glucose ; metabolism ; Glucose Transporter Type 1 ; metabolism ; Glucose Transporter Type 4 ; metabolism ; Hep G2 Cells ; Humans ; Insulin ; pharmacology ; Insulin Resistance ; Liver ; metabolism ; Mice

OBJECTIVETo probe into an effective therapy for chronic pain of cervical intervertebral disc.

METHODSSixty-six cases of chronic pain of cervical intervertebral disc were randomly divided into an acupoint sticking group and an acu puncture group, 33 cases in each group. The acupoint sticking group was treated with acupoint sticking therapy, and the acupuncture group with acupuncture. Tianding (LI 17), Futu (LI 18), Dazhui (GV 14), Tianzong (S 11), etc. were selected in the two groups. After treatment for 3 therapeutic courses, their therapeutic effects were observed.

RESULTSThe effective rate was 93.5% in the acupoint sticking group and 72.4% in the acupuncture group, the former being better than the latter (P<0.05); in a 3-month following-up survey, the effective rate was 90.3% in the acupoint sticking group and 65.5% in the acupuncture group, the former being better than the latter (P<0.05).

CONCLUSIONThe acupoint sticking therapy has an satisfactory therapeutic effect on chronic pain of cervical intervertebral disc.

Acupuncture Points ; Acupuncture Therapy ; Administration, Cutaneous ; Adult ; Aged ; Back Pain ; drug therapy ; therapy ; Cervical Vertebrae ; drug effects ; Chronic Disease ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Humans ; Intervertebral Disc ; drug effects ; Male ; Middle Aged ; Young Adult

OBJECTIVETo study the distribution and expressions of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) in the III-IV stage of pressure ulcer wound, and to explore their correlation with ulceration.

METHODSForty-one patients hospitalized in the two Affiliated Hospital of Wenzhou Medical College from June 2010 to March 2012 were recruited, including twenty-one patients with 23 pressure ulcer of stage III-IV, 14 acute injury patients, and 6 donors of normal skin. Samples harvested from the 41 patients through surgery were divided into four groups, including pressure ulcer centre group (n = 23), pressure ulcer margin group (n = 23), acute wound group (n = 14), and normal skin group (n = 6). The histological changes in wounds were observed after HE staining. The distribution of collagen fiber in wound was observed with Masson staining. Expressions of VEGF and bFGF in wounds were detected with immunohistochemical staining. Data were processed with independent samples t test and paired samples t test.

RESULTS(1) In the two pressure ulcer groups, large number of inflammatory cells were found in aggregation; the expression of collagen fiber was decreased or disappeared; the positive expressions of VEGF and bFGF were mainly located in fibroblasts and endothelial cells. The expression levels of VEGF and bFGF were respectively 100 ± 39, 132 ± 46 in pressure ulcer centre group, and 228 ± 48, 299 ± 80 in pressure ulcer margin group. The differences between the two pressure ulcer groups were statistically significant (with t values respectively 13.497 and 13.020, P values below 0.01). (2) In acute wound group, a large number of fibroblasts but a small amount of collagen fibers were observed; the positive expressions of VEGF and bFGF were mainly located in fibroblasts, with respective expression levels of 292 ± 59 and 443 ± 194, which were significantly higher than those of the two pressure ulcer groups (with t values from 2.370 to 11.570, P < 0.05 or P < 0.01). (3) In normal skin group, structure of tissue was appropriate, and abundant collagen fibers were observed; the expression levels of VEGF and bFGF were respectively 45 ± 18 and 54 ± 22, which were significantly lower than those of the other three groups (with t values from 3.983 to 14.087, P values all below 0.01).

CONCLUSIONSIn contrast with those of the acute wounds, the expression levels of VEGF and bFGF are significantly decreased in the pressure ulcer wound at stage III-IV. It may be closely correlated with the decrease or cessation of the synthesis of collagen fiber.

Adult ; Aged ; Aged, 80 and over ; Female ; Fibroblast Growth Factor 2 ; metabolism ; Humans ; Male ; Middle Aged ; Pressure Ulcer ; metabolism ; Vascular Endothelial Growth Factor A ; metabolism ; Wound Healing