Burnout, Professional ; Humans

To prepare virus-like particles containing FluA/B mRNA as RNA standard and control in Influenza RNA detection, the genes coding the coat protein and maturase of E. coli bacteriophage MS2 were amplified and cloned into D-pET32a vector. Then we inserted 6 histidines to MS2 coat protein by QuikChange Site-Directed Mutagenesis Kit to construct the universal expressing vector D-pET32a-CP-His. In addition, the partial gene fragments of FluA and FluB were cloned to the down-stream of expressing vector. The recombinant plasmid D-pET32a-CP-His-FluA/B was transformed to BL21 with induction by IPTG. The virus-like particles were purified by Ni+ chromatography. The virus-like particles can be detected by RT-PCR, but not PCR. They can be conserved stably for at least 3 months at both 4 degrees C and -20 degrees C. His-tagged virus-like particles are more stable and easier to purification. It can be used as RNA standard and control in Influenza virus RNA detection.
Escherichia coli ; genetics ; metabolism ; Influenza A virus ; genetics ; metabolism ; Influenza B virus ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; RNA, Viral ; genetics ; metabolism ; Recombinant Fusion Proteins ; genetics ; metabolism ; Ribonucleases ; chemistry ; Virion ; genetics ; metabolism

Objective To evaluate the curative value of the tauroursodeoxycholic acid (TUDCA) and Danning table (DN) in the prevention of bile duct stones recurrence after ERCP. Methods 210 patients with Choledocholithiasisby ERCP were randomly divided into 3 groups. The patients in the control group were not given any medicine. The patients in the TUDCA group took TUDCA every night. The patients in the DN group took Danning tablets of three times a day. The course of medication and followed up for the patients was 24 months after the operation. All the patients would be examined regularly by Biliary color doppler ultrasound and MRCP. The recurrence rates of Choledocholithiasis, cholesterol saturation index and serum lipid were recorded and compared statistically between the three groups. Results In total, 190 patients completed the treatment and follow-up according to the experimental design. The recurrence rates of the control, TUDCA and DN group were 17.46%(11/63), 6.34% (4/63) and 10.94% (7/64). The recurrence rates in the TUDCA and DN group were significantly lower than those the control group (P < 0.05).The recurrence rate in the TUDCA group was significantly lower than that in the DN group; On the tenth days after the operation, the CSI in the TUDCA were significantly lower than those the control group and the DN group (P < 0.05). There was no statistically significant difference between the DN group and the control group (P > 0.05); To the patients without recurrence, the level of TC, LDL in the TUDCA group were significantly lower than those the control group and and the DN group (P < 0.05). The level of HDL in the TUDCA group were significantly higher than those the control group and and the DN group (P < 0.05). There was no statistically significant difference on the level of HDL between the DN groupand the control group (P > 0.05). Conclusion The application of TUDCA and DN for patients with Choledocholithiasis after ERCP can effectively reduce the recurrence rates, the curative effect of TUDCA more than DN.

Objective To evaluate the clinical value of combined detection of T cell receptor rear-rangement excision circles ( TRECs) and CD31+ regulatory T ( Treg) cells for accessing the recent thymic output in patients with chronic hepatitis B. Methods Four groups involving 135 subjects were set up in this study as follows: mild chronic hepatitis B ( Mild CHB, n=35 ) , moderate chronic hepatitis B ( Moderate CHB, n=35 ) , severe chronic hepatitis B ( Severe CHB, n=35 ) and healthy control ( HCs, n=30 ) groups. CD4+CD25+Treg cells in these subjects were sorted out using magnetic cell separation. The ratio of peripheral CD31+Treg cells to Treg cells in each group was analyzed by flow cytometry. Real-time PCR was performed to detect TRECs in CD4+CD25+Treg cells. The percentages of CD3+, CD4+ and CD8+T cell sub-sets were also measured. Results The ratios of CD31+Treg/Treg cells and the numbers of TRECs in pe-ripheral blood of the Moderate CHB and Severe CHB groups were significantly lower than those of the Mild CHB and HCs groups (P<0. 05), while no statistical difference was found between the mild CHB and HC groups (P>0. 05). No significant difference in the percentages of CD3+, CD4+ or CD8+ T cell subsets was observed between the four groups (P>0. 05). CD31+ Treg/Treg cell ratio had a positive correlation with the number of TRECs (r=0. 551, P=0. 014). Conclusions Both CD31+Treg/Treg cell ratio and the number of TRECs were reduced in the peripheral blood of patients with moderate or severe CHB. CD31+Treg/Treg cell ratio and the number of TRECs were positively correlated and could be used as new indices to evaluate recent thymus output.

To observe and analyze therapeutic effect of valsartan combined beraprost sodium on patients with hypertension (EH) complicated early renal injury (ERI).Methods : A total of 480 EH + ERI patients treated in our hospital in near two years were randomly and equally divided into valsartan group and combined treatment group (received valsartan combined beraprost sodium) , both groups were treated for three months .Levels of blood pressure , renal function related indexes were compared between two groups before and after treatment .Results :Compared with before treatment , after three months , there were significant reductions in levels of blood pressure , serum creatinine , urine β2 microglobulin and D‐dimer , and significant rise in creatinine clearance rate (Ccr) in two groups , P＝0.001 all.Compared with valsartan group after treatment , there were significant reductions in levels of blood pressure [ (130. 92 ± 5.92)／(80.18 ± 6.69) mmHg vs.(120. 93 ± 6. 53)／(69.98 ± 6.32) mmHg] , serum creatinine [ (93.92 ± 10. 49) μmol／L vs.(83. 14 ± 11. 03) μmol／L] , urine β2 microglobulin [ (385.41 ± 35.54) μg／L vs.(362.65 ± 26.59) μg／L] and D‐dimer [ (1. 75 ± 0.44) mg／L vs.(1. 01 ± 0.11) mg／L] , and significant rise in Ccr [ (63.22 ± 7. 66) ml／min vs.(79.13 ± 8.83) ml／min] in combined treatment group , P＝0.001 all.Conclusion :Compared with valsartan monotherapy , valsartan combined beraprost sodium can more significantly reduce blood pressure and protect renal function in hypertensive patients with early renal injury .

The aim of the present study was to investigate the effect of lipoxin A4 (LXA4)pretreatment on cognitive function of aged rats after global cerebral ischemia reperfusion,and to explore its possible mechanism.Thirty-six aged male Sprague-Dawley rats were randomly divided into three groups (n=12 each):sham-operation group (S group),global cerebral ischemia reperfusion group (I/R group) and LXA4-pretreatment group (L group).The rat model of global cerebral ischemia reperfusion was established by occlusion of the bilateral common carotid artery with hypotension.The cognitive function of rats was determined by a step-down type passive avoidance test and Morris Water Maze test on the third day after reperfusion.Rats were sacrificed after Water Maze test and the pathological changes ofhippocampal CA1 region were observed and the related inflammatory mediators were determined.As compared with S group,the escape latency in I/R group was prolonged from the first day to the fifth day,while that in L group was prolonged from the first day to the third day.The retention time in I/R group and L group in the first quadrant was shortened.The reaction time,frequency of reaction mistake and frequency of escape mistake in I/R group increased,and the latent period shortened.The frequency of escape mistake in L group increased,and the damage in the hippocampal CA1 region of I/R group and L group was obvious.The levels of S-100β,TNF-α,IL-lβ,IL-10 and NF-κB in I/R group and L group increased.As compared with I/R group,the escape latency in L group was shortened from the first day to the fifth day,and the retention time in the first quadrant prolonged.The reaction time,frequency of reaction mistake and frequency of escape mistake in L group decreased,and the latent period prolonged.The damage in the hippocampal CA1 region of L group was alleviated as well.The levels of S-100β,TNF-α,IL-1β and NF-κB in L group decreased,and those of IL-10 increased.It can be concluded that LXA4 pretreatment can improve the cognitive function in aged rats after global cerebral ischemia reperfusion probably by inhibiting the inflammatory reaction.

OBJECTIVE: The stability of ketamine in biological samples was studied under different storage temperature and time. METHODS: The rabbits were given intragastric administration of ketamine with a dose of 150 mg/kg and were sacrificed after 30 minutes. Blood, liver, kidney and brain of the rabbits were stored at room temperature (between 18 degrees C and 24 degrees C) and -20 degrees C. The specimens were analyzed at different times by GC-MS and GC-NPD. RESULTS: At -20 degrees C, the concentration of ketamine decreased in all of samples (P < 0.05) within 30 days. The concentration of ketamine increased in all of samples stored at room temperature after 5 days(P < 0.05). CONCLUSION The stability of ketamine in biological samples stored at -20 degrees C was better than that at room temperature. The samples suspected containing ketamine should be stored at -20 degrees C and should be tested as soon as possible.
Animals ; Brain/metabolism* ; Cryopreservation ; Disease Models, Animal ; Drug Stability ; Female ; Forensic Toxicology ; Gas Chromatography-Mass Spectrometry/methods* ; Hydrogen-Ion Concentration ; Ketamine/poisoning* ; Kidney/metabolism* ; Liver/metabolism* ; Male ; Rabbits ; Specimen Handling/methods* ; Temperature ; Time Factors

OBJECTIVETo explore the signal transduction mechanisms of apoptosis in renal tubular epithelial cells in diabetic rats with fluctuant high blood glucose.

METHODSHealthy SD rats were randomly divided into 3 groups: normal control group (A), stable high blood glucose group (B) and fluctuant high blood glucose group (C). Diabetic rats were induced by intraperitoneal injection of streptozotocin (STZ, 65 mg/kg), and the fluctuant high blood glucose animal model was induced by intraperitoneal injection of ordinary insulin and glucose at different time point every day. The superoxide dismutase (SOD) activity and the content of malonaldehyde (MDA) in renal tissue homogenate were detected with colorimetry. The protein expression of Nox4 and JNK were examined by immunohistochemistry and Western blot. Apoptosis was assessed by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labelling (TUNEL).

RESULTSAfter 12 experimental weeks, significantly increased cell apoptosis, up-regulation of Nox4 and P-JNK expression in renal tubular epithelial cells were observed in B and C groups compared with those in A group. The MDA content increased and SOD activity decreased in renal tissue in B and C groups. Above effects were more obviously shown in C group.

CONCLUSIONFluctuant high blood glucose induced more apoptosis of renal tubular epithelial cell than stable high blood glucose in diabetic kidney, which might be related to the activation of JNK signal transduction pathway.

Animals ; Apoptosis ; Blood Glucose ; metabolism ; Diabetes Mellitus, Experimental ; metabolism ; pathology ; Epithelial Cells ; metabolism ; Kidney Tubules ; cytology ; MAP Kinase Kinase 4 ; metabolism ; MAP Kinase Signaling System ; Male ; Malondialdehyde ; metabolism ; NADPH Oxidase 4 ; NADPH Oxidases ; metabolism ; Rats ; Rats, Sprague-Dawley ; Superoxide Dismutase ; metabolism

The aim of the present study was to investigate the expression changes of three steroidogenic enzymes in the polycystic ovary syndrome (PCOS). Thirty Sprague-Dawley (SD) rats were randomly divided into normal control (NC) group and PCOS group. PCOS rat model was established by DHEA injection. The serum levels of progesterone, estrogen and testosterone were measured by immunoradioassay or enzyme immunoassay. The cellular distributions of 3β-hydroxy steroid dehydrogenase (3β-HSD), 17β-hydroxy steroid dehydrogenase (17β-HSD) and cytochrome P450 aromatase (P450arom) in ovaries were detected by immunohistochemistry. The expression levels of 3β-HSD, 17β-HSD and P450arom were detected by RT-PCR and Western blot. The results showed that the serum levels of estrogen and testosterone of PCOS group were significantly higher than those of the NC group. There was no significant difference of serum progesterone level between the PCOS and NC groups. Compared with the NC group, the PCOS group showed increased mRNA and protein expressions of both 3β-HSD and 17β-HSD, as well as reduced P450arom mRNA and protein expressions. These results suggest that 3β-HSD and 17β-HSD, but not P450arom, may participate in the ovarian hormonal regulation in the present rat model of PCOS.
17-Hydroxysteroid Dehydrogenases ; metabolism ; 3-Hydroxysteroid Dehydrogenases ; metabolism ; Animals ; Aromatase ; metabolism ; Disease Models, Animal ; Estrogens ; blood ; Female ; Polycystic Ovary Syndrome ; enzymology ; Progesterone ; blood ; Rats ; Rats, Sprague-Dawley ; Testosterone ; blood

OBJECTIVETo examine the role of glutaminergic neurons in the transmission and integration of the sweat taste information in the brain stem and the amygdala.

METHODSConscious Sprague-Dawley rats were subjected to oral sweet taste or water (control) stimulations. The activated neurons were identified by detecting c-Fos expression in taste-related brain areas, and the glutaminergic neurons by detecting vesicular glutamate transpoter-3 (VGLUT3).

RESULTSCompared with control group, the rats with oral sucrose solution stimulation exhibited significantly increased c-Fos-expressing and double-labeled neurons in the nucleus of the solitary tract (NST), the parabrachial nucleus (PBN) and the amygdala.

CONCLUSIONNeurons in the NST, PBN and amygdala are activated after oral sweet taste stimulation. The sweet taste perception at different levels in the CNS is partly mediated by glutamate.

Amygdala ; physiology ; Animals ; Brain Stem ; physiology ; Glutamic Acid ; metabolism ; Male ; Neurons ; metabolism ; physiology ; Proto-Oncogene Proteins c-fos ; metabolism ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; Solitary Nucleus ; cytology ; physiology ; Sucrose ; administration & dosage ; metabolism ; Taste Perception ; physiology ; Vesicular Glutamate Transport Proteins ; metabolism