An HPLC-UV method has been developed for the determination of valibose, miglitol, voglibose and acarbose, the four anti-diabetic drugs. The separation was accomplished successfully by using reversed phase chromatography (Prevail carbohydrate column, 250 mm x 4.6 mm, 5 microm) with a gradient acetonitrile-phosphate buffer solution (pH 8.0) at a wavelength of 210 nm. Furthermore, the method of a high-performance liquid chromatography coupled with ESI-MS in positive ionization mode has been established. These two methods were successfully applied to the assay and qualitative detection of four alpha-glucosidase inhibitors in the potential counterfeit anti-diabetic drugs.

Neutrophils play a critical role in ventilation-induced lung injury. This study was aimed to investigate the characteristics of neutrophils influx in lungs induced by high tidal volume ventilation. Anaesthetized rats were randomly divided into low tidal volume ventilation group (Vt: 7 mL/kg, LV group) or high tidal volume ventilation group (Vt: 42 mL/kg, HV group) (n=40 in each). Rats in each group were ventilated for 0, 60, 90, 120 and 240 min. The wet/dry lung weight ratio (W/D) was measured. The levels of macrophage inflammatory protein-2 (MIP-2) and tumor necrosis factor-α (TNF-α), and the activity of myeloperoxidase (MPO) were detected by enzyme-linked immunosorbent assay (ELISA). The number of neutrophils in bronchoalveolar lavage fluid (BALF) was counted after Wright's staining, and the percentage of netrophils in lung tissues calculated. Histopatholgical examination was used to observe the changes of lung tissues after different ventilations. The results showed that the W/D weight ratio was increased, and the levels of MIP-2 and TNF-α significantly enhanced in HV group at 90, 120 and 240 min. Neutrophils in BALF and the neutrophil percentage in lung tissues were also elevated at 120 and 240 min, which coincided with the enhanced activity of MPO in HV group. The lung injury was significantly related with the ventilation time and the infiltration of neutrophils in lungs in HV group. In conclusion, in ventilation-induced lung injury, neutrophil infiltration is present in a time-dependent manner and associated with the aggravated lung injury. Pulmonary structural damage may be the main reason for ventilation-induced lung injury.

Objective To study the effect of Kanglaite combined with comprehensive therapy on advanced non-small cell lung cancer.Methods Sixty-one patients with advanced non-small cell lung cancer were randomly divided into treatment group ( n=31 ) and control group ( n=30 ).Both groups were given comprehensive therapy.Treatment group were additionally treated with intravenous injection of 200 ml Kanglaite.Clinical efficacy,quality of life,pain relief and adverse reactions of the two groups were observed.Results ( 1 ) Quality of life was improved in 20 cases (64.5％ ),stabled in 8 cases (25.8％),declined in 3 cases ( 9.7％ ) of treatment group,and in the control group there were 9 cases ( 30.0％ ) improved,9 cases ( 30.0％ ) stabilized,12 cases (40.0％ ) declined respectively.Quality of life in treatment group was higher than in control group ( U=2.91,P＜0.01 ).( 2 ) Pain relief:the number of patients with complete remission,partial remission,no change,and progression were 5 cases ( 16.1％ ),16 cases ( 51.6 ％ ),16 cases (51.6％ ),6 cases (19.4％) and 4 cases (12.9％ ) in treatment group,and in control group they were 2 cases(6.7％ ),9 cases (30.0％ ),11 cases(36.7％ ) and 8 cases(26.7％ ) respectively.The effect of treatment on pain relief in treatment group was better than that in control group ( U=2.32,P＜0.05 ).(3) Clinical efficacy:in the treatment group there were 12 cases (38.7％) with partial remission,14 cases (45.2％) stabilized,and 5 cases (16.1％ ) progressed,and in control group the numbers were 8 cases (26.7％ ),8 cases (26.7％ ) and 14 cases (46.7％ ) respectively.The clinical efficacy in treatment group was better than that in the control group( U=2.04,P＜0.05).(4) There were significant difference on the change of white blood cell count and gastrointestinal reactions Ⅲ and Ⅳ degrees between treatment group and contrl group [22.6％ (7/31) vs.53.3％(16/30),x2=6.139 P＜0.05;19.4％ (6/31) vs.46.7％ (14/30),x2=5.161,P＜0.05].Conclusion Kanglaite injection combined with comprehensive therapy can improve clinical efficacy of therapy for advanced non-small cell lung cancer,reduce the toxic adverse reaction,protect immunity system and improve the quality of life of patients.

Objective To explore pathological characterization of the enlarged contrast enhanced area of hepatocellular carcinoma ( HCC) on contrast enhanced ultrasonography ( CEUS). Methods Thirty-five HCC patients underwent conventional ultrasonography and CEUS before hepatectomy. The tumor sizes were measured on conventional ultrasonography,CEUS and postoperative resected specimen at the largest section. The infiltration depth of the HCC was observed under microscope in the peripheral area of the tumors. HCC with the size of contrast enhancement area on CEUS 0. 3 cm larger than that on conventional ultrasonography was defined as group A, and the others as group B. The pathological character of the tumor and tissues surrounding the tumor were analyzed and compared between the two groups. Results Among 35 HCC, 40% (14/35) HCC increased in size on CEUS compared with ultrasonography. Of them,9 tumors with increased area on CEUS were confirmed to be invasive HCC on pathology. Using the enlarged contrast enhanced area on CEUS as the diagnostic criteria of invasive HCC, the sensitivity, specificity, accuracy, positive predictive value and negative predictive value were 90%, 80%, 82.9%, 64.3%, 95. 2%, respectively. Conclusions The contrast enhanced area on CEUS can reflect the infiltrative tumor size more accurately compared with conventional ultrasonography.

Background and purpose: The literature on dose-volume parameters and pneumonitis is extensive. The results are inconsistent, both for the best predictive metrics and significant comorbid factors. This study aimed to investigate a prospective functional equivalent uniform dose (fEUD) with perfusion single photon emission computed tomography (SPECT) images as predictors of radiation pneumonitis (RP) in patients undergoing curative radiotherapy (RT). Methods: Functional lung imaging was performed using SPECT for perfusion imaging. Perfusion factors were defined as the mean percentile perfusion levels of the 4 areas, top to 75%, 75% to 50%, 50% to 25%, 25% to 0%, re-spectively. fEUD was calculated from perfusion factors and standard dose-volume parameters extracted from treatment planning computed tomography (CT) scans. Total lung (TL), ipsilateral (IL) and contralateral lung (CL) volumes minus gross tumor volume (GTV), whole-lung V5, V20, whole lung fEUD, IL and CL fEUD, and general equivalent uniform dose (gEUD) were analyzed to evaluate correlations between RP using Common Terminology Criteria for Adverse Events (CTCAE) version 4.03. Statistical significance was defined as P<0.05. Results: A total of 15 patients treated with intensity modulated RT or 3D conformal RT were analyzed, grades≥3 RP were observed in 6 patients. There was only a trend toward significance for unilateral (UL) fEUD of higher dose side (P=0.007). Whole-lung V5, V20 were almost identical between patients who developed pneumonitis and patients who did not, as the values were below the recommended thresholds from published papers. Unilateral fEUDs were linear with unilateral gEUDs (t=0.815, P=0.000). Conclusion: SPECT-based equivalent uniform dose appears to be a better predictor of RP compared to stan-dard dose-volume parameters. Planning constraints should aim to keep unilateral fEUD below 21 Gy.

Case recruitment of large-scale clinical trials should be strictly checked in quality and quantity for it is the key to clinical trial. This study discusses the main difficulties and countermeasures in the case recruitment of large sample, multi-center clinical trials according to the national research project "Myocardial Infarction Secondary Prevention Study in Traditional Chinese Medicine".

Objective:To observe the difference of the human telomeres RNA component (hTERC) genes’amplification in the cervical tissue by applying the environment-friendly fixative poly hydroxy acrylic acid and the transparent dewaxing solution Van-clear separately or jointly to replace the traditional fixative 4% (volume fraction)neutral buffered formalin and the conventional transparent dewaxing solu-tion xylene in the use of fluorescence in situ hybridization (FISH)for detection.Methods:In the study, 255 cases of cervical tissue specimens submitted by the Department of Gynecology in Zhongshan Boai Ho-sipital were collected from Mar.2013 to Apr.2015.Four samples were taken from the same lesion site. All the cases were divided into 4 groups and named group A,B,C,and D.Group A used 4% neutral buffered formalin fixed and xylene dewaxing to make slices.Group B used poly hydroxy acrylic fixed and xylene dewaxing to make slices.Group C used 4% neutral buffered formalin fixed and Van-clear trans-parent to make slices.Group D used poly hydroxy acrylic fixed and Van-clear transparent dewaxing to make slices.The amplification of hTERC genes in the four groups of cervical specimens was also detected by FISH technique.Results:When the hTERC genes were detected by FISH method under the fluore-scence microscope,it was obvious that the tissue profile and the background of group A,B,C and D were all clear.The probe was fixed in the accurate position so that the bright red or green fluorescence signals were easily found in these four groups.Compared with the positive rate of group A,there was no statistical significance in that of group B,C and D (P>0.05).At the same time,the coincidence rate of the FISH results was high,which showed that the new environment-friendly reagent had no significant difference in the detection of cervical hTERC genes by FISH technique.Conclusion:It is possible for the environment-friendly reagent poly hydroxy acrylic acid and Van-clear to replace 4%neutral buffered for-malin and xylene separately or jointly to detect the cervical hTERC genes by adopting FISH technique.

Background and purpose: Adequate tissue ifxation, transparent dewaxing is an important step of hematoxylin eosin (HE) staining and lfuorescence in situ hybridization (FISH) in detection of breast cancerHER-2 gene. The purpose of this study was to make a comparison between poly hydroxyl acrylic acid which is an environmen-tally friendly ifxation liquid and 4% neutral buffered formaldehyde in tissue ifxation for HE staining and FISH to detect theHER-2 gene in the breast cancer tissue sections. The study aimed to evaluate the feasibility of replacing 4% buffered formaldehyde, a traditional ifxation liquid, with the poly hydroxyl acrylic acid, an environmentally friendly ifxation lfuid.Methods:This project was performed on tissue samples collected from 69 cases of breast cancer, 41 cases of breast ifbroadenoma, 40 cases of uterine leiomyoma, 25 cases of cervical tissue, 25 cases of placenta obtained from the outpatient and inpatient departments of Zhongshan Boai Hospital from Mar. 2011 to Jan. 2015, from each of which two samples were drawn and two blocks of each specimen were divided into two groups randomly. Then one group was ifxed with 4% neutral buffered formaldehyde and made into 200 sections by HE while the other group was ifxed with poly hydroxyl acrylic acid and made into another 200 sections. The slice level of the two groups was determined by the staining condition of the sections, and SPSS 19.0 was employed to compare the excellent and good rate of HE staining. Additional 69 sections were produced with two groups of breast invasive ductal carcinoma tissues, and SPSS 19.0 was used to detect the ampliifcation ofHER-2 gene by FISH.Results:First, the number of best-quality slices stained with HE ifxed separately by poly hydroxyl acrylic acid and 4% neutral buffered formaldehyde was 155 and 166, respectively. The number of excellent pieces was 41 and 33, respectively, while the number of mediocre pieces was 3 and 1 with bad pieces being 1 and 0, respectively. The excellent and good rates of HE staining were 98% and 99.5%, respectively. There was no significant difference between the two groups (χ2=1.33,P>0.05).Second, the positive rates of the tis-sue slices by FISH ifxed separately by poly hydroxyl acrylic acid and 4% neutral buffered were 26.09% and 23.19%, respectively. There was no signiifcant difference between the two groups (χ2=0.50,P>0.05).Conclusion:The results obtained with HE staining and FISH using poly hydroxyl acrylic acid as a ifxation liquid are not signiifcantly different from those using 10% neutral buffered formaldehyde. Therefore, poly hydroxyl acrylic acid meets the requirements of environmental protection, and thus has the potential to be promoted and widely used.

Neutrophils play a critical role in ventilation-induced lung injury. This study was aimed to investigate the characteristics of neutrophils influx in lungs induced by high tidal volume ventilation. Anaesthetized rats were randomly divided into low tidal volume ventilation group (Vt: 7 mL/kg, LV group) or high tidal volume ventilation group (Vt: 42 mL/kg, HV group) (n=40 in each). Rats in each group were ventilated for 0, 60, 90, 120 and 240 min. The wet/dry lung weight ratio (W/D) was measured. The levels of macrophage inflammatory protein-2 (MIP-2) and tumor necrosis factor-α (TNF-α), and the activity of myeloperoxidase (MPO) were detected by enzyme-linked immunosorbent assay (ELISA). The number of neutrophils in bronchoalveolar lavage fluid (BALF) was counted after Wright's staining, and the percentage of netrophils in lung tissues calculated. Histopatholgical examination was used to observe the changes of lung tissues after different ventilations. The results showed that the W/D weight ratio was increased, and the levels of MIP-2 and TNF-α significantly enhanced in HV group at 90, 120 and 240 min. Neutrophils in BALF and the neutrophil percentage in lung tissues were also elevated at 120 and 240 min, which coincided with the enhanced activity of MPO in HV group. The lung injury was significantly related with the ventilation time and the infiltration of neutrophils in lungs in HV group. In conclusion, in ventilation-induced lung injury, neutrophil infiltration is present in a time-dependent manner and associated with the aggravated lung injury. Pulmonary structural damage may be the main reason for ventilation-induced lung injury.
Animals ; Lung ; physiopathology ; Lung Injury ; physiopathology ; Male ; Neutrophil Infiltration ; physiology ; Neutrophils ; physiology ; Rats ; Rats, Sprague-Dawley

Objective To compare two different methods to detect the differences of gene mutation rate, sensitivity, specificity and coincidence rate of epidermal growth factor receptor (EGFR) in non-small-cell lung cancer (NSCLC) so as to assess the clinical value of qRT-PCR method and its environmental-friendly technologyplatforms.One uses environmental fixative poly hydroxyl acrylic acid and green transparent liquid dewaxing Van-clear alone or in combination to replace the traditional fixative 4％ (volume fraction) neutral buffered formalin and the traditional transparent dewaxing liquid xylene in application of quantitative real-time polymerase chain reaction (qRT-PCR).The other uses traditional reagents in direct sequencing.Methods We selected 91 cases of primary NSCLC specimens resected between May 2013 and March 2016 in Zhongshan Bo`ai Hospital and Zhongshan Hospital of Traditional Chinese Medicine.Five samples were taken from the same tumor lesion.We used a random number table to randomly divide these samples into Groups A, B , C, D, and E.Group A received direct sequencing method in detection of EGFR gene mutations.Besides, during the experiment, 4％ neutral buffered formalin was used for fixing, and xylene transparent dewaxing was used to make slices for DNA extraction dewaxing.Group B received qRT-PCR method to detect EGFR gene mutations.Meanwhile, during the experiment, 4％ neutral buffered formalin was used for fixing, and xylene transparent dewaxing was used to make slices for DNA extraction dewaxing.Group C received qRT-PCR method in detection of EGFR gene mutations.At the same time, during the experiment, polyhydroxy acrylic acid was used for fixing, and xylene transparent dewaxing was used to make slices for DNA extraction dewaxing.Group D received qRT-PCR method to detect EGFR gene mutations.In the meantime, 4％ neutral buffered formalin was used for fixing, Van-clear transparent dewaxing was used to make slices for DNA extraction dewaxing.Group E received qRT-PCR method in detection of EGFR gene mutations.In addition, during the experiment, polyhydroxy acrylic acid was used for fixing, and Van-clear transparent dewaxing was used to make slices for DNA extraction dewaxing.In addition, during the experiment, polyhydroxy acrylic acid was used for fixing, and Van-clear transparent dewaxing was used to make slices for DNA extraction dewaxing.The mutations of Exons 18, 19, 20, and 21 in EGFR genes were respectively determined in the five groups of NSCLC.Results ① Groups B, C, D, E and A did not significantly differ in the percentage of people with mutations or target site mutation rates of EGFR genes in NSCLC (P> 0.05).② The detection results of EGFR target site mutation in Groups B, C, D, E and A had good sensitivity, strong specificity, and high compliance rate.Conclusion The green transparent liquid dewaxing Van-clear alone or in combination to replace the traditional fixative 4％ neutral buffered formalin and the traditional transparent dewaxing liquid xylene in the application of qRT-PCR so as to detect EGFR gene mutations in NSCLC has good consistent results compared with the method that uses traditional reagents in direct sequencing.It has the significance and value in clinical application.