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 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 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

AIM: To provide the morphological evidence for the compound carbomer building high intraocular pressure model successfully.? METHODS: Totally 50 SD rats were divided into experimental group 40 rats, blank group 10 rats by random number method. The rats in experimental group were randomly divided into 3 groups after the model was successful. Experimental model of high intraocular pressure was sacrificed at 1, 2 and 3wk to observe its pathological structure change and ultrastructure change.? RESULTS: The high intraocular pressure of experimental model in 1, 2 and 3wk all showed the optic nerve and retinal damage. It was to see the optic nerve axon disappear, disorder arrangement of myelin sheath, periodic dissolve or demyelinating degeneration, glial cell proliferation. It showed the cells disordered arrangement of retina, the outer nuclear layer became thick, the inner and outer plexiform layer become thick, the kernel layer became thick, air bubbles, the numbers of ganglion cells reduced, ganglion cells and nerve fiber layer edema, microglia proliferation, vascular membrane capillaries expansion, inflammatory cells appearing. It was to see the retinal ganglion cells layer with microglia proliferation under the electron microscope, ganglion cells structure fuzzy, organelles structures disappear, cell apoptosis, mitochondrial swelling, cytoplasm vacuoles degeneration, membrane plate of outer segment fracture or dissolved. And the damage degree was proportional to the forming time of high intraocular pressure.? CONCLUSION: The morphology change of high intraocular pressure model about the retin and optic nerve proves that it is successful building the model through anterior chamber injection of compound carbomer solution.