OBJECTIVETo explore the underwater decompression schedule for 100 m Trimix conventional diving operations and evaluate its safety through a simulated rabbits Trimix conventional diving.

METHODSAccording to the Haldane theory, the assumed time units, the classification of tissue compartments, the nitrogen super-saturation safety coefficient and the selection of methods used for the calculation of the simulated 100 m Trimix conventional diving schedule were properly selected, and the calculating method for the dive decompression schedule was thus firmly established. In our experiments, five tissue compartments were selected during the calculation of decompression schedule: 5 min, 10 min, 20 min, 40 min and 75 min, and the nitrogen super-saturation safety coefficient was calculated by 1.6. Eight New Zealand rabbits were performed a simulated 100 m Trimix dive program which was established according to the Haldane theory, and eight rabbits for intact group. The tissues wet/dry ratio and ethology were detected and observed before and after the simulated diving to evaluate the safety of decompression schedule.

RESULTSBy using the developed underwater decompression schedule, abnormal ethology changes in rabbits could not be observed after compression and decompression to the surface; and the tissues wet/dry ratio of simulated diving rabbits had no significant changes compared with the intact group (P > 0.05).

CONCLUSIONThe decompression schedule calculated by Haldane theory seemed to be safe and reliable, the diving breathing gas concentration did not cause oxygen toxicity and nitrogen narcosis among the dive rabbits, and dive efficiency was greatly improved by using enriched oxygen gas in UPTD safety range during decompression.

Animal Experimentation ; Animals ; Decompression ; Diving ; Helium ; Nitrogen ; Oxygen ; Rabbits

BACKGROUND:Many types of mammalian cells aggregate and display three-dimensional multicellular spheroids when they are in normal physiological conditions. In order to observe and explore cellular natural states, many researchers try to use spherical cellculture in vitro, a common three-dimensional culture pattern.
OBJECTIVE:To use three different methods for spherical culture in vitro of adipose-derived stem cells and to observe their biological features.
METHODS:Adipose-derived stem cells were confirmed by the analysis of the markers for cellphenotypes as wel as adipogenic and osteogenic differentiation potential assays. Three different methods of sphere cultures were used as fol ows:(1) ultra low attachment culture;(2) hanging-drop culture and (3) Eppendorf tube culture. The sphere formation was compared among above three methods. We used Imagej to calculate mean areas of these spheres. And we used Viability/Cytotoxicity Assay Kit for Animal Live&Dead cells to detect their vitality.
RESULTS AND CONCLUSION:(1) Adipose-derived stem cells were confirmed by the analysis of the markers for cellphenotypes, CD29, CD44, CD59 were positive, as wel as adipogenic and osteogenic differentiation potential assays were positive. The conventional monolayer cultures of adipose-derived stem cells showed spindle and cloning growth within three passages. (2) Ultra low attachment culture, hanging-drop culture, Eppendorf tube culture al could elicit adipose-derived stem cells spherical growth. However, spherical size, shape and uniformity differed depending on cellnumbers, culture time and spherical culture methods. The ultra low attachment culture was comparatively difficult to control spherical shape and uniformity of adipose-derived stem cells. But hanging-drop culture and Eppendorf tube culture were able to form even cellspheres. (3) Spherical formation of adipose-derived stem cells using our three methods displayed good cellvitality.

Objective:To compare the therapeutic and adverse effects of chemotherapeutic regimen based on three drugs (taxol+carboplatin/cisplatin+etoposide) and two drugs (carboplatin/cisplatin+etoposide) on the combined small cell lung cancer (CSCLC). Methods:A retrospective study was conducted based on the data of 62 CSCLC patients who were admitted to and treated at Tianjin Medical University Cancer Institute and Hospital between July 2000 and April 2013. Of the 62 patients, 19 received the three-drug regi-men and 43 received the two-drug regimen. All patients received at least two cycles of chemotherapy and completed follow-up proce-dures. For each patient, the therapeutic efficacy was evaluated every two cycles, and toxicity was evaluated every cycle. Results:The response rates between the three-drug and two-drug groups were statistically significant (90%vs. 53%, P=0.033). However, no statisti-cal differences were observed in the disease control rate between the two groups (100% vs. 86%, P=0.212). The three-drug regimen could induce a better median progression-free survival compared with the two-drug regimen, but with no statistical significance (10.5%vs. 9.8%, P=0.484). Similarly, no statistical differences were noted in the median overall survival between the three-drug and two-drug groups (24.0%vs. 17.5%, P=0.457). The incidence rates of grade IV bone marrow depression and the termination of the original regi-men owing to severe adverse reactions were both significantly higher in the three-drug group than in the two-drug group (26.3% vs. 7.0%, P=0.036;31.6%vs. 14.7%, P=0.004). Conclusion:The two-drug regimen had almost the same survival rate and lower toxicity compared with the three-drug regimen. When using the TEP/TCE regimen, a close attention should be focused on its adverse reactions. The findings of this work showed that the two-agent regimen should be one of the standard treatments for CSCLC.

Objective To investigate X‐ray ,CT and MRI features of synovial chondromatosis of the temporomandibular joint (TMJ) .Methods X‐ray ,CT and MRI features of eight patients of synovial chondromatosis of TMJ with histo‐pathologically con‐firmed were analyzed retrospectively .X‐ray examination and CT scanning were performed in all eight patients .Routine MRI scanning was performed in six patients and contrast‐enhanced MRI scanning was performed in two patients synchronously .Results Tumors occured unilaterally in all eight cases ,which occured on the right TMJ in six cases and on the left side in two cases .On X‐ray films , widen joint space and calcificated loose bodies occured in all eight cases .On CT scanning ,cystic‐solid mixed mass around the joint and calcificated loose bodies occured in all eight cases .On MR scanning ,multiple nodular long T1 and short T2 signal occured in six cases . Arthroedema and synovial hyperplasia with iso T1 and iso or slightly long T2 signal in six cases .On contrast‐enhanced MR ,homoge‐neous enhancement occurred in svnovial tissue and the edge of loose bodies in two cases .Conclusion The synovial chondromatosis of TMJ owns typical imaging features .The imaging findings can serve as a reference to improve diagnosis of synovial chondromatosis of TMJ .

Background The construction of tissue-engineered corneal endothelium and corneal endothelial cells (CECs) therapy need abundant seed cells,so how to culture a large amount of CECs with high viability and original cell properties is an urgent issue to be solved.Objective This study was to establish three-dimensional spheroid culture of CECs and explore the cellular biological characteristics.Methods Primary rabbit CECs were isolated with trypsin and subcultured.Low attachment and shaking culture was applied to form CECs spheres.Cultured cells were identified under the inverted microscope.The surface features of the cells were examined under the scanning electron microscope.The viability of the cells were assayed by acridine orange (AO) staining and CCK-8 kit.Then CECs spheres were incubated to 6-well plate for 1 week,and immunofluorescence staining was used to identify the expression of zonula occludens-1 (ZO-1) and Na+/K+-ATPase in the cells.The cell proliferation value of spheroid culture method was compared with that of regular culture method.Results CECs grew into aggregation after cultured with the hexagonal or polygonal shape and tight connection among the cells.The cells converged into single layer and slabstone-like arrangement 1 week later.Cells migrated out of the CECs sphere and formed an uneven spherical surface.The living cells showed green fluorescence for AO with the survival rate 90％.The absorbance (A450) of the cells was 1.524±0.013 and 1.265 ±0.021 in the spherical culture group and conventional culture group,respectively,showing a significant difference between them (t =-3.436,P=0.010).The positive cells of ZO-1 and Na+/K+-ATPase showed the green fluorescence for FITC on cell membrane and blue fluorescence for DAPI on cell nucleus.Conclusions Spherical culture method maintains a high viability and proliferation ability of the cells and remains phenotype of CECs,which is superior to conventional culture method.This culture method provides better seeding CECs for the establishment of tissue engineering cornea endothelial layer and CECs therapy.

In order to investigate the sleep quality and influencing factors in patients with Parkinson's disease (PD),201 PD patients were enrolled and underwent extensive clinical evaluations.Subjective sleep evaluation was assessed using the Pittsburgh Sleep Quality Index (PSQI),and the Epworth Sleepiness Scale (ESS).It was found that poor sleep quality (77.11％) and excessive daytime sleepiness (32.34％) were commonly seen in PD patients and positively correlated with disease severity.Then 70 out of the 201 PD patients and 70 age-and sex-matched controls underwent a polysomnographic recording.The parameters were compared between PD group and control group and the influencing factors of sleep in PD patients were analyzed.The results showed that sleep efficiency (SE) was significantly decreased (P＜0.01),and sleep latency (SL) and the arousal index (AI) were increased (P＜0.05) in the PD group as compared with those in the control group.SE and total sleep time (TST) were positively correlated with the Hoehn and Yahr (H&Y) stage.There was significant difference in the extent of hypopnea and hypoxemia between the PD group and the control group (P＜0.05).Our results indicate that PD patients have an overall poor sleep quality and a high prevalence of sleep disorder,which may be correlated with the disease severity.Respiratory function and oxygen supply are also affected to a certain degree in PD patients.

OBJECTIVETo explore the differential gene expressions of chronic hepatitis B (CHB) of Gan depression Pi deficiency syndrome (GDPDS) and Pi-Wei damp-heat syndrome (PWDHS).

METHODSThe ulnar venous blood was withdrawn from healthy subjects (26 cases), patients of GDPDS (35 cases) and PWDHS (34 cases) on an empty stomach. The total RNA was extracted using Trizol method. The differential genes were detected using Aglient expression profile chip and screened using randomized variance model. The results were analyzed using GO, Pathway, GeneBank, NCBI, and Geneontology.

RESULTSThere were 125 differential genes between CHB patients of GDPDS and those of PWDHS (including 66 up-regulated genes and 59 down-regulated genes), mainly involving in functions of transmembrane transport, response to selenium ion, and regulation of calcium ion-dependent exocytosis. The signal pathway participated in mainly includes cell adhesion molecules, calcium ion signaling pathway, leukocyte trans-endothelial migration. We present gene co-expression networks to find 9 interactions among genes (LOC340508, HIST2H2BE, MPL, FLJ22536, TUBA8, NT5M, EGFL7, PTPRF, TSPAN33), which were mainly involved in immune response, cell growth, DNA damage, signal transduction, inflammatory reaction, and so on.

CONCLUSIONSThe differential expression genes existed between CHB patients of GDPDS and those of PWDHS, indicating that Chinese medicine syndrome classification has its own basis for gene expression profile. The genomics research method is expected to provide an objective basis for Chinese medicine syndrome typing.

Adult ; Case-Control Studies ; Female ; Gene Expression ; Hepatitis B, Chronic ; diagnosis ; genetics ; Humans ; Male ; Medicine, Chinese Traditional ; methods ; Oligonucleotide Array Sequence Analysis ; Transcriptome

Objective To establish high-performance liquid chromatography with electrochemical detector(HPLC-ECD) method for the determination for metanephrine and normetanephrine in 24 h urine, and provide a superior test for the diagnostic of pheochromocytomas over plasma/urine catecholamine.Methods MCX solid-phase cartridge was used for extraction of metanephrine and normetanephrine,HPLC-ECD was used for their measurements.The intra-assay CVs,interassay CVs and recoveries of metanephrine and normetanephrine were also calculated.104 hypertensive patients without pheochromocytomas and 5 pheochromocytomas patients were selected in this study.The concentrations of metanephrine and normetanephrine were compared with the plasma and 24h urinary catecholamines concentrations.Results The intra-assay CV,inter-assay CV and recovery of metanephrine were 5.9%, 7.5%,91.1% respectively;the intra-assay CV,inter-assay CV and recovery of normetanephrine were 6.3%,6.6%,88.5%,respectively.The MN,NMN,plasma CA and urine CA of all pheochromocytomas patients were positive.MN and NMN were negative in controls,while plasma CA and urine CA are false positive in 15 patients and 14 patients in controls,respectively.Conclusions The study establish a fast and accurate method for quantification of metanephrine and normetanephrine in 24 h urine by HPLC-ECD.These findings also prove that it is the best biochemical assays for pheochromocytomas at present.

Objective To observe the clinical effect of etomidate combined with propofol on laparoscopic cholecystectomy.Methods Eight patients undergoing laparoscopic cholecystectomy cho sen from March 2015 to April 2016 in our hospital,falling into ASA Ⅰ or Ⅱ grades,randomly divided into four groups,20 in each.Group A was administered of propofol (2 mg/kg);group B etomidate (0.3 mg/kg);group C etomidate (0.15 mg/kg) combined with propofol (1 mg/kg);group D etomidate (0.1 mg/kg) combined with propofol (1.5 mg/kg).Systolic blood pressure,diastolic blood pressure and heart rate of the four group were compared at the following six time points:before anesthesia induction(T0),1 min before intubation (T1),the intubation moment (T2),1 min after intubation (T3),5 min after intubation (T4),15 min after intubation (T5).The serum of four groups were collected at the time before anesthesia induction (T0),30 min after anesthesia induction (Tb),2 h after induction of anesthesia (Tc) and 24 h after anesthesia induction(Td).Aldosterone,cortisol and adrenocorticotropic hormone in serum were tested by radioimmunoassay.The incidence of adverse reactions such as injection pain and myoclonus during induction,and postoperative nausea and vomiting of the four groups were recorded and compared.Results Compared with T0,HR,SBP at T1,T4 and T5 were significantly decreased in group A (P＜0.05).SBP at T2 were significantly increased in group B (P＜0.05),and SBP at T1 and T5 were significantly decreased in group C (P＜0.05).There was no significant difference of HR at T1,T2,T3,T4 and T5 in group B and C.Compared with T0,HR,SBP at T1,and T5 were significantly decreased in group D (P＜0.05).There was no significant difference in hemodynamics among the four groups.Compared with T0,the levels of Cor and ALD at Tb and Tc were decreased in group B and C (P＜0.05).And the level of ACTH of B and C in Tc were significantly increased (P＜0.05).Compared with group A,the levels of Cor and ALD of group B and C were significantly decreased at Tb and Tc (P＜0.05).Compared with group B,the levels of Cor and ALD of group C and D showed less decrease at Tb and Tc (P＜0.05).And compared with group C,the levels of Cor and ALD of group D showed less decrease at Tb and Tc (P＜0.05).Conclusion Etomidate combined with propofol induced can reduce the side effects induced by propofol or etomidate alone,which is an ideal way of anesthesia induction.

BACKGROUNDGalectin-3 is the most recently identified advanced glycosylation end products (AGEs) binding protein. This study aimed to investigate the effects of AGEs and rosiglitazone on the expression and secretion of galectin-3 in cultured human renal mesangial cells (HRMCs).

METHODSHRMCs were incubated with different concentrations of AGE-bovine serum albumin (BSA) (0, 50, 100, 200, and 400 mg/L) for different time (0, 24, 36, 48, and 72 hours), and exposed to AGE-BSA in the presence of different concentrations of rosiglitazone (1, 10, and 100 micromol/L). The mRNA and protein expression of galectin-3 in HRMCs were analyzed by reverse transcription polymerase chain reaction (RT-PCR) and Western blotting. The culture medium of HRMCs was collected and concentrated, and the content of galectin-3 in the medium was detected by Western blotting.

RESULTSBoth RT-PCR and Western blotting revealed that AGE-BSA up-regulated the expression of galectin-3 in HRMCs in a concentration- (P < 0.05) and time-dependent (P < 0.05) manner compared with the control. Compared with the control, AGE-BSA elevated the content of galectin-3 in the culture medium of HRMCs time- and concentration-dependently (P < 0.05, respectively). Both protein and mRNA expression of galectin-3, and its content in the medium of HRMCs exposed to different concentrations of rosiglitazone in the presence of AGE-BSA were increased compared with those of cells exposed to AGE-BSA alone (P < 0.05). Rosiglitazone increased the expression and secretion of galectin-3 in a dose-dependent manner (P < 0.05).

CONCLUSIONSAGEs up-regulates the expression and secretion of galectin-3 in HRMCs. Rosiglitazone further enhances the upregulation of galectin-3 in HRMCs induced by AGEs, which suggests that rosiglitazone may play a role of reno-protection via up-regulation of galectin-3.

Blotting, Western ; Cell Line ; Galectin 3 ; genetics ; secretion ; Glycation End Products, Advanced ; pharmacology ; Humans ; Hypoglycemic Agents ; pharmacology ; Mesangial Cells ; drug effects ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Serum Albumin, Bovine ; pharmacology ; Thiazolidinediones ; pharmacology