OBJECTIVETo apply the Wright-Giemsa stain in micronucleus test and to explore the stain outcomes of Wright-Giemsa dye of various proportions and staining times.

METHODSUse Wright-Giemsa dye, Wright dye (staining time 3 min) and Giemsa dye (staining time 5 min) to stain HepG2 and then observe the staining effect. The Wright-Giemsa dye was applied under 5 different proportions (3:1-1:3) and different staining times (1, 3, 5, 10, 15 min).

RESULTSAfter stained for 3-5 min with the proportion ratio of 3:1 of Wright-Giemsa dye, the HepG2 cells showed much better staining outcomes compared with the single stain of either Wright or Giemsa.

CONCLUSIONSWright-Giemsa stain can be used in cell micronucleus test to obtain good staining outcomes.

Azure Stains ; Coloring Agents ; Hep G2 Cells ; Humans ; Micronucleus Tests ; Staining and Labeling ; methods

BACKGROUND: Platelet endothelial cell adhesion molecule-1 (PECAM-1), also known as CD31, is mainly distributed in vascular endothelial cells. Studies have shown that PECAM-1 is a very significant indicator of angiogenesis, and has been used as an indicator for vascular endothelial cells. The present study aimed to explore the relationship between the expression of PECAM-1 and the degree of acute lung injury (ALI) and fibrosis in paraquat (PQ) induced lung injury in rabbits. METHODS: Thirty-six adult New Zealand rabbits were randomly divided into three groups (12 rabbits in each group) according to PQ dosage: 8 mg/kg (group A), 16 mg/kg (group B), and 32 mg/kg (group C). After PQ infusion, the rabbits were monitored for 7 days and then euthanized. The lungs were removed for histological evaluation. Masson staining was used to determine the degree of lung fibrosis (LF), and semi-quantitative immune-histochemistry analysis to determine the expression of PECAM-1. Pearson's product-moment correlation analysis was performed to evaluate the relationship between the expression of PECAM-1 and the extent of lung injuries expressed by ALI score and degree of LF. RESULTS: Rabbits in the three groups showed apparent poisoning. The rabbits survived longer in group A than in groups B and C (6.47±0.99 days vs. 6.09±1.04 days vs. 4.77±2.04 days) (P<0.05). ALI score was lower in group A than in groups B and C (8.33±1.03 vs. 9.83±1.17 vs. 11.50±1.38) (P<0.05), and there was statistically significant difference between group B and group C (P=0.03). LF was slighter in group A than in groups B and C (31.09％±2.05 ％ vs. 34.37％±1.62％ vs. 36.54％±0.44％) (P<0.05), and there was statistically significant difference between group B and group C (P=0.026). The PEACAM-1 expression was higher in group A than in groups B and C (20.31％±0.70％ vs. 19.34％±0.68％ vs. 18.37％±0.46％) (P<0.05), and there was statistically significant difference between group B and group C (P=0.017). Pearson's correlation analysis showed that the expression of PECAM-1 was negatively correlated to both ALI score (Coe=–0.732, P=0.001) and degree of LF (Coe=–0.779, P<0.001). CONCLUSIONS: The PECAM-1 expression significantly decreases in New Zealand rabbits after PQ poisoning, and the decrease is dose-dependent. The PECAM-1 expression is negatively correlated with ALI score and LF, showing a significant role in the development of lung injuries induced by PQ.

BACKGROUNDThe coagulation function in carcinoma patients is abnormal, but in renal cell carcinoma the extent and relationships of coagulation function remain unclear. This study retrospectively investigated the relationships between coagulation function, clinical stage and metastasis in patients with renal cell carcinoma.

METHODSA total of 350 consecutive patients admitted to our Urology Department from 2004 to 2010 were diagnosed with renal cell carcinoma by histopathologic examination and were included in this study. A total of 231 cases of renal benign tumors were considered as the control group. Fibrinogen, prothrombin time, activated partial thromboplastin time and international normalized ratio were evaluated in all subjects. Tumor size, clinical stage, lymph node metastasis, and distant metastasis were evaluated using radiologic imaging, intraoperative findings, and histological studies.

RESULTSThe preoperative plasma fibrinogen levels of patients with renal cell carcinoma ((383.9 ± 146.7) mg/dl) were significantly higher than those of the control group ((316.7 ± 62.0) mg/dl) (P < 0.01). We divided the renal cell carcinoma group into stages Ia, Ib, II, III, and IV. The fibrinogen values were (315.6 ± 64.6) mg/dl, (358.3 ± 91.1) mg/dl, (465.6 ± 164.7) mg/dl, (500.0 ± 202.1) mg/dl, and (585.8 ± 179.7) mg/dl, respectively. There were no significant differences in fibrinogen values between stage Ia and control groups. However, results of other stages showed significant differences when compared to control group values (P < 0.01). Using the cutoff value of 440 mg/dl, which defines hyperfibrinogenemia, plasma fibrinogen levels had a positive predictive value of 39.8% and a negative predictive value of 93.3% for predicting distant metastasis, with a sensitivity of 64.7% and specificity of 83.3%.

CONCLUSIONSPreoperative plasma fibrinogen levels are elevated in patients with renal cell carcinoma with distant metastasis or lymph node metastasis. Potential metastasis is more likely if the tumor size larger than 4 cm. Increased preoperative plasma fibrinogen levels, especially hyperfibrinogenemia, may be an indicator of metastasis.

Adult ; Aged ; Blood Coagulation ; physiology ; Carcinoma, Renal Cell ; metabolism ; pathology ; physiopathology ; Female ; Fibrinogen ; metabolism ; Humans ; Kidney Neoplasms ; metabolism ; pathology ; physiopathology ; Lymphatic Metastasis ; physiopathology ; Male ; Middle Aged ; Neoplasm Metastasis ; physiopathology ; Neoplasm Staging ; Retrospective Studies ; Thromboplastin ; metabolism

BACKGROUND:To invent a novel cardiopulmonary resuscitation (CPR) time point recorder to synchronously and automatically record the time and to identify its effectiveness in humans. METHODS:A CPR time point recorder was invented after the doctors were familiar with the traditional Utstein recovery registration mode and mastered the registration time points required. The progress of CPR was simulated. The standard and correct times were recorded, and the doctors performing the recovery collected the data about the times using our CPR time point recorder or the memory registration mode. RESULTS:The deviation times were 21.4±24.7 seconds for the memory group and 3.57±4.58 seconds for CPR time point recorder group. The deviation of times increased significantly depending on the increase of the operation items in the memory group. A similar phenomenon was found in the timer group but with a smaller difference (P<0.01). CONCLUSION: A CPR time point recorder could reduce the deviation of operate-time, especially after a long-time operation, and for procedures with more operating items, compared with the memory mode. It was a more advantageous and accurate method for the Utstein registration.

OBJECTIVEIntravenous anesthetics, such as propofol, are widely used in general anesthesia. Neurodegeneration and neurocognitive impairment after exposure to propofol in neonatal rats have raised concerns regarding the safety of pediatric anesthesia. We examined the effects of neonatal propofol exposure on brain cell viability, as well as expression of hippocampal survivin and Caspase-3 mRNA and protein.

METHODSOne hundred male Sprague-Dawley rats aged 7 d that were weighed 10-15 g were randomly divided into 4 groups (n = 25 each group). Group A: the rats were injected with no drugs. Group B: the rats were intraperitoneally injected with 50 mg/kg propofol. Group C: the rats were first intraperitoneally injected with 50 mg/kg propofol and another 50 mg/kg propofol was used when the dynamic response of rats appeared again. Group D: the rats were first intraperitoneally injected with 50 mg/kg propofol and another 50 mg/kg propofol was used three times once the dynamic response of rats appeared. To study the effects of propofol exposure on respiratory and metabolic function, arterial blood was aspirated from the left ventricle of neonatal rats 2 h after discontinuation of propofol. pH, PaO(2), PaCO(2), HCO(3)(-), BE and SaO(2) were detected by blood gas analyzer. Moreover, to examine the effects of propofol exposure on short-term cellular viability, the ultrastructure of neurons was observed by transmission electron microscope and Fluoro-Jade B (FJB) staining was performed to examine neuronal degeneration in hippocampal CA1 region of neonatal rats. Survivin and Caspase-3 mRNA and protein expression in hippocampus were detected by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) and Western blotting 2 h after discontinuation of propofol.

RESULTSThe time of anesthesia maintaince in newborn rats was the longest in Group D and the time of anesthesia maintaince in Group C was longer than that in Group B. Two hours after discontinuation of propofol, pH, PaO(2), PaCO(2), HCO(3)(-), BE and SaO(2) of arterial blood in rats were not significantly different among groups A, B, C and D (P > 0.05). The structure of hippocampal neurons was normal in Group A and Group B while 100 mg/kg propofol resulted in nuclear blebbing and 200 mg/kg propofol led to nuclear fragmentation, chromatin condensation and apoptotic bodies. Cellular degeneration, as measured by Fluoro-Jade B staining, significantly increased in hippocampal CA1 region in the anesthesia groups compared with littermates in the no anesthesia group. FJB-positive stained degenerative neurons in groups B, C and D were (2.5 ± 1.3), (7.1 ± 2.3) and (9.4 ± 2.6), which were different from that in Group A (0.6 ± 0.3) (P < 0.05). Moreover, the number of FJB-positive neurons was the highest in Group D, that in Group C was more than that in Group B. At the same time point, apoptosis was measured by expression of Caspase-3 and Survivin mRNA and protein in hippocampus of rats. Caspase-3 mRNA in groups A, B and C was (0.78 ± 0.12), (0.84 ± 0.17) and (0.89 ± 0.19), while Caspase-3 protein in groups A, B and C was (0.22 ± 0.05), (0.26 ± 0.07) and (0.21 ± 0.06). Survivin mRNA in groups A, B and C was (0.56 ± 0.12), (0.58 ± 0.15) and (0.53 ± 0.16), while Survivin protein in these 3 groups was (0.24 ± 0.07), (0.21 ± 0.05) and (0.23 ± 0.06). Compared with that in Group A, Caspase-3 and Survivin mRNA and protein were not significantly different among Group B and Group C (P > 0.05). However, Caspase-3 mRNA and protein in Group D were (1.21 ± 0.14) and (0.42 ± 0.12), which were higher than that in the other 3 groups (P < 0.05). Survivin mRNA and protein in Group D were lower than that in the other 3 groups (P < 0.05).

CONCLUSIONSA high dose of propofol exposure may destroy the structure of neurons, induce neurodegeneration, increase Caspase-3 activity and inhibit survivin expression in hippocampus of newborn rats in vivo.

Anesthetics, Intravenous ; administration & dosage ; pharmacology ; Animals ; Animals, Newborn ; Blood Gas Analysis ; Caspase 3 ; genetics ; metabolism ; Dose-Response Relationship, Drug ; Gene Expression Regulation ; drug effects ; Hippocampus ; drug effects ; metabolism ; Injections, Intraperitoneal ; Male ; Microtubule-Associated Proteins ; genetics ; metabolism ; Neurons ; metabolism ; pathology ; Propofol ; administration & dosage ; pharmacology ; RNA, Messenger ; genetics ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo explore the effect of different culture media on viability and expression of tau protein in organotypic hippocampal slice.

METHODSBrain slices (400 microm) from 1, 2, 4, and 8 week-old Wistar rats were prepared and cultured in minimum essential medium (MEM) or Dulbecco's modified eagle medium: nutrient mixture (DMEM/F12) medium respectively for 21 days. Viability of the slices was measured by lactate dehydrogenase (LDH) assay and expression of tau protein was detected by Western blot.

RESULTSThe viability of the slices was not influenced significantly by the two different culture media, while the expression level of tau protein was significantly higher in DMEM/F12 than in MEM (P < 0.05), especially in the slices from 2 and 4 week-old rats.

CONCLUSIONThe slices from 2 or 4 week-old rat hippocampi and DMEM/F12 medium may be the preferred choice for tau associated researches. An ideal Alzheimer's disease model may be established based on the results of these researches.

Animals ; Culture Media ; Hippocampus ; growth & development ; metabolism ; L-Lactate Dehydrogenase ; biosynthesis ; Organ Culture Techniques ; methods ; Rats ; Rats, Wistar ; tau Proteins ; biosynthesis ; genetics

This study is to reveal the correlation between CNVs of HMGR, SQS1, beta-AS gene and genuineness of liquorice. Real-time PCR was used to detect the copy number of HMGR, SQS1, beta-AS gene of liquorice. According to the results, the range of the copy number variation of HMGR gene was between 1 and 3, the copy number of SQS1 gene was 1 or 2, and the copy number of beta-AS gene was only 1. On the basis of the copy number of HMGR, SQS1 and beta-AS gene, there were five groups, type A (2 + 1 + 1), type B (1 + 1 + 1), type C (3 + 2 + 1), type D (2 + 2 + 1) and type E (3 + 1 + 1). There were two types, type A and type B, in Hangjinqi of Inner Mongolia, and the ratio of A to B was 1:1.3. There were also two types, type A and type B, in Chifeng of Inner Mongolia, and the ratio of A to B was 3:1. There were four types, type A, type B, type C and type D, in Yanchi of Ningxia province, and the ratio of A to B was 1:5.1. There were three types, type A, type B and type E, in Minqin of Gansu province, and the ratio of A to B was 2:1. So CNVs mainly existed in the liquorice from Ningxia and Gansu provinces. While the genetic background of liquorice from Hangjinqi of Inner Mongolia was stabilized. The results of the experiment proved that the correlation between CNVs and origins was one of the reasons of genuineness of liquorice.
China ; DNA Copy Number Variations ; DNA, Plant ; genetics ; Farnesyl-Diphosphate Farnesyltransferase ; genetics ; Glycyrrhiza uralensis ; enzymology ; genetics ; Hydroxymethylglutaryl CoA Reductases ; genetics ; Intramolecular Transferases ; genetics ; Real-Time Polymerase Chain Reaction

OBJECTIVETo obtain large quantities of well differentiated urinary bladder transitional epithelial cells for used as the seed cells in bladder tissue engineering, and evaluate the cytocompatibility of silk fibroin film with the transitional cells in vitro to assess the possibility of tissue-engineered urinary organ construction.

METHODSThe urinary bladder transitional epithelial cells were isolated from the bladders of New Zealand rabbits and cultured in vitro as the seed cells, whose morphology was observed and the specific protein (cytokeratin) expression identified by immunofluorescence assay. The cells were seeded in 96-well plates at 1 x 10(>4)/ml and incubated with silk fibroin film leaching solution or culture medium (negative control). MTT assay was performed to determine the cell proliferation rates of the wells and evaluate the cytotoxicity and cytocompatibility of the silk fibroin film.

RESULTSThe isolated urinary bladder transitional epithelial cells reached confluence after 9-10 days of culture, which showed positive staining for immunocytochemistry with monoclonal antibody against cytokeratin. The absorbance of the cells culture in the presence of silk fibroin film leaching solution averaged 0.424-/+0.020, 0.996-/+0.118 and 1.285-/+0.048 after at 24, 72 and 120 h of cell culture, and that of the negative control group at the time points was 0.419-/+0.030, 1.105-/+0.098 and 1.228-/+0.052, respectively, showing no significant difference between the two groups (P>0.05).

CONCLUSIONSilk fibroin film has good cytocompatibility with rabbit urinary bladder transitional epithelial cells, and may serve as good scaffold material for urologic tissue engineering.

Animals ; Biocompatible Materials ; Cell Culture Techniques ; Cells, Cultured ; Epithelial Cells ; cytology ; Fibroins ; chemistry ; Keratins ; analysis ; Rabbits ; Tissue Engineering ; Urinary Bladder ; cytology

Objective To evaluate the enhanced magnification endoscopy in the diagnosis of Barrett esophagus,and to explore the relationship between mucosal surface patterns and pathological epithelial types of Barrett esophagus.Methods Enhanced magnification endoscopy was performed 'after spraying 2%-3% acetic acid on the surface of distal esophagus in 40 Barrett esophagus patients.Mucosal specimen were biop- syed.Results According to the mucosal types of Toyoda in 2003,there were three mucosal types:Ⅰ dot pat- tern 7(17.5%),5 of 7(71.4%)fundie type,Ⅱ reticular pattern 24(60.0%),16 of 24(66.7%)fundic type,Ⅲ cerebroid/villous 9(22.5%),intestinal metaplasia or dysplasia.Conclusion Enhanced magnifi- cation endoscopy helps to identify areas with intestinal metaplasia and dysplasia,and is useful in the diagno- sis of Barrett esophagus.

BACKGROUNDThe feasibility and safety of endoscopic thyroidectomy were evaluated by an approach of systematic review of published studies in the past decade.

METHODSA database searching was performed on MEDLINE, Cochrane Database of Systematic Reviews, American College of Physicians Journal Club, Database of Abstracts of Reviews of Effects, Cochrane Central Register of Controlled Trials. Both comparative and non-comparative studies about endoscopic thyroidectomy were selected and analyzed. For the comparative studies, RevMan 4.2 was used for statistical analysis; and for the non-comparative studies, data analysis was performed by SPSS 13.0.

RESULTSSeven comparative studies involving 367 patients (video-assisted thyroidectomy (VAT), 174 patients; conventional thyroidectomy (CT), 193 patients) were included in VAT-CT group. Age, gender, operative types, and pathological diagnosis were similar. Compared with CT, the mean operative time for VAT was significantly longer (VAT, 80.0 minutes; CT, 61.9 minutes, P < 0.01), but the postoperative hospital stay was shorter (VAT, 1.7 days; CT, 2.5 days, P < 0.01). The complication rate for VAT was 6.9%, while that for CT was 9.3% (P = 0.35). Three studies analyzed the postoperative pain and cosmetic evaluation, and indicated that the VAT group was superior to the CT group, but there was no significant difference after a meta-analysis. Three comparative studies involving 273 patients (totally endoscopic thyroidectomy (TET), 145 patients; CT, 128 patients) were included in TET-CT group and the results generally resembled that of VAT-CT group. There were 18 and 14 non-comparative studies reporting the results of VAT and TET, respectively. The mean operative time for VAT was 76.8 minutes compared with 135.8 minutes for TET. The postoperative hospital stay was 1.8 and 3.8 days for VAT and TET respectively. The rates of conversion to open surgery for VAT and TET were similar (VAT, 2.8%; TET, 3.9%, P = 0.105). The complication rate for VAT was 8.6%, while that for TET was 3.5% (P < 0.01).

CONCLUSIONSThe feasibility and safety of endoscopic thyroidectomy were initially verified and accepted, and it should be considered as a valid option, offering some advantages to patients in terms of cosmetic results and postoperative distress.

Adult ; Endoscopy ; methods ; Humans ; Pain, Postoperative ; drug therapy ; Thyroidectomy ; adverse effects ; methods ; Video-Assisted Surgery