Objective To study the expression of protein kinase B (Akt/PKB) isoforms in the gastrocnemius muscle of rats at different ages. Methods The expression levels of mRNA and protein of three Akt isoforms in the gastrocnemius muscle of 30-month-old and 6-month-old rats were detected respectively by RT-PCR and Western blotting. Data were analyzed statistically. Results (1) The levels of Akt1 and Akt2 mRNA in the 30-month-old rats was significantly higher than those in the 6-month-old rats (t=7.124, P=0.000; t=2.598, P=0.021), however, no significant difference was found in the level of Akt3 mRNA between the two groups (t=0.460, P=0.653) . (2) The level of Akt-Thr308 phosphorylation in the 30-month-old rats was significantly lower than that in the 6-month-old rats (t=-9.861, P=0.000), while the level of Akt2 protein in the 30-month group was significantly higher than that in the younger rats (t=7.522, P=0.000). No significant differences were detected in the levels of Akt1 and Akt3 proteins between the two groups (t=0.469, P=0.646; t=0.058, P=0.955). Conclusion The expression levels of three Akt isoforms in the gastrocnemius muscle of rats change with age, suggesting that the three isoforms of Akt have different functions in the gastrocnemius muscle metabolism of rats at different ages.

This study aimed to analyze the etiology of the encephalitis outbreak in Longyan, Fujian Province, China in 2010, in order to provide valuable information for this prevention and control of this disease. Pathogens were confirmed from cerebrospinal fluid samples with fluorescent RT-PCR, virus isolation (RD cells), and neutralization tests. Then, the VP1 fragments or whole genome nucleotide sequences were determined for four virus strains using PCR. Homology was assessed using the MegAlign software, and a phylogenetic evolutionary tree was drawn using Mega 4.0 software. The results confirmed that the etiology of the outbreak was the ECHO6 intestinal virus, and the nucleotide sequence of the VP1 segment indicated that the C2 subtype was responsible. The genome sequence consisted of 7407 nucleotides, and resembled the genome of other ECHO and CoxB viruses with homology levels of 78.5%-87.3%. The encephalitis outbreak in Longyan in 2010 was caused by the ECHO6 C2 subtype intestinal virus, and its complete genome sequence length is similar to the standard strain (U16283) with a sequence homology of 80.4%.
Child, Preschool ; China ; epidemiology ; Disease Outbreaks ; Echovirus 6, Human ; classification ; genetics ; isolation & purification ; Echovirus Infections ; epidemiology ; virology ; Encephalitis ; epidemiology ; virology ; Female ; Humans ; Infant ; Male ; Molecular Sequence Data ; Phylogeny

OBJECTIVETo assess the efficacy of tendons of minimally invasive therapy (TMIT) combined drug therapy by comparing it with treatment by drug therapy alone on patients with knee osteoarthritis (KOA).

METHODSTotally 60 KOA patients were assigned to the treatment group and the control group according to random digit table, 30 in each group. Patients in the control group took Hydrochloric Acid Glucosamine Capsule and Celecoxib Capsule. Patients in the treatment group additionally received TMIT. The treatment course for all was 4 weeks. Scores for visual analogue scale (VAS) and the Western Ontario and McMaster Universities (WOMAC) Osteoarthritis Index were observed and recorded at week 1 and 4 after treatment by acupotomology mirror.

RESULTSCompared with before treatment, improvement was shown in VAS score, pain and stiffness degrees, activities and functions, and WOMAC scores at week 1 and 4 after treatment in all patients with statistical difference (P < 0.05). Besides, better effect was shown in the treatment group (P < 0.05).

CONCLUSIONSTMIT combined drug therapy could relieve KOA patients' pain, stiffness and joint activities, elevate the overall efficacy. TMIT was easily operated with less injury.

Celecoxib ; Drug Therapy, Combination ; methods ; Humans ; Knee Joint ; Osteoarthritis, Knee ; drug therapy ; Pain ; Pain Measurement ; Tendons ; Treatment Outcome

OBJECTIVETo determine dopamine and its metabolites during in vivo cerebral microdialysis by routine high performance liquid chromatography with electrochemical detection.

METHODSMicrodialysis probes were placed into the right striatum of Wistar rat brains and perfused with Ringer's solution at a rate of 1.5 microL/min. A reverse phase HPLC with electrochemistry was used to assay DA, DOPAC, and HVA after cerebral microdialysates were collected every 20 minutes from awake and freely moving rats. In order to identify the reliability of this method, its selectivity, linear range, precision and accuracy were tested and the contents of DA, DOPAC, and HVA in rat microdialysates were determined.

RESULTSThe standard curve was in good linear at the concentration ranging from 74 nmol/L to 1.5 micromol/L for DOPAC (r2=0.9996), from 66 nmol/L to 1.3 micromol/L for DA (r2=1.0000) and from 69 nmol/L to 1.4 micromol/L for HVA (r2=0.9992). The recovery of DOPAC (0.30, 0.77, 1.49 micromol/L), DA (0.26, 0.69, 1.32 micromol/L), and HVA (0.27, 0.71, 1.37 micromol/L) was 82.00+/-1.70%, 104.00+/-4.00%, 98.70+/-3.10%; 92.30+/-1.50%, 105.30+/-2.30%, 108.00+/-2.00%; 80.00+/-7.80%, 107.69+/-8.00%, and 108.66+/-3.10%, respectively at each concentration. Their intra-day RSD was 3.3%, 3.4%, and 2.5%, and inter-day RSD was 4.2%, 2.3%, and 5.6%, respectively. The mean extracellular concentrations of DOPAC, DA, and HVA in rat brain microdialysates were 10.7, 2.4, and 9.2 micromol/L (n=6), respectively.

CONCLUSIONThe findings of our study suggested that the simple, accurate and stable method can be applied to basic researches of diseases related to monoamines neurotransmitters by cerebral microdialysis in rats.

Animals ; Brain ; metabolism ; Chromatography, High Pressure Liquid ; methods ; Dopamine ; metabolism ; Electrochemistry ; methods ; Microdialysis ; Rats ; Reference Standards ; Sensitivity and Specificity

An S-naproxen (S-NAP) molecularly imprinted monolithic stationary phase (MIMSP) with specific recognition for S-NAP and naproxen (NAP) was prepared by in situ technique,utilizing 4-vinylpridine (4-VP) as a function monomer,ethylene glycol dimethacrylate (EDMA) as a cross-linking agent,and low-polar solvents (toluene and dodecanol) as porogenic solvents.The selectivity of the polymers for S-NAP and NAP was evaluated by high performance liquid chromatography (HPLC).The binding characteristics were tested by Scatchard analysis.Racemic NAP could be specifically separated to some extent.At the same time,NAP could be separated from ibuprofen under optimized conditions.Scatchard analysis showed that two classes of binding sites existed in the S-NAP-imprinted polymers,with their dissociation constants estimated to be 1.045 and 5.496 μM,respectively.The results demonstrate that S-NAP and NAP can be recognized specifically on the obtained MIMSP.

Objective To study the effect of antiapoptosis factors PED/PEA-15 and XIAP on prostate cancer cells(PC-3)apoptosis.Methods The expressions of XIAP and PED/PEA-15 in prostate cancer cells(PC-3)were respectively assayed using the RT-PCR technique.XIAP and PED/ PEA-15 specific siRNA vectors were designed and constructed and then were transiently cotransfected into PC-3 cells under induction of liposome.The effects of siRNA vectors on PED/PEA-15 and XIAP transcription were assayed by RT-PCR technique,and the effect of XIAP and PED/PEA-15 on cancer cell apoptosis were determined by flow cytometry and microscope observation.Results PED/PEA- 15 and XIAP were both highly expressed in PC-3 cells.Enzyme digestion analysis and DNA sequencing confirmed that the PED/PEA-15 and XIAP-specific siRNA expression vectors were constructed successfully.The designed siRNA sequences of PED/PEA-15 and XIAP could specifically inhibit their transcription.The PC-3 cells which were cotransfected with PED/PEA-15 and XIAP- specific siRNA vectors were more sensitive to doxorubicin.The apoptosis rate of cotransfected cells was significantly increased.Conclusions PED/PEA-15 and XIAP might be involved in the development of prostate cancer.

Objective To explore an algorithm to define the threshold value for tumor contouring on 18F-fluorodexyglucose (FDG) PET imaging. Methods A National Electrical Manufacturing Association (NEMA)NU 2 1994 PET phantom with 5 spheres of different diameters were filled with 18F-FDG. Seven different sphere-to-background ratios were obtained and the phantom was scanned by Discovery LS 4. For each sphere-to-background ratio, the maximum standardized uptake value ( SUVmax ) of each sphere, the SUV of the border of each sphere ( SUVborder ), the mean SUV of a 1 cm region of background (SUVbg) and the diameter (D) of each sphere were measured. SPSS 13.0 software was used for curve fitting and regression analysis to obtain the threshold algorithm. The calculated thresholds were applied to delineate 29 pathologically confirmed lung cancer lesions on PET images and the obtained volumes were compared with the volumes contoured on CT images in lung window. Results The algorithm for defining contour threshold is TH% = 33.1% + 46.8% SUVbg/SUVmax + 13.9%/D ( r = 0.994) by phantom studies. For 29 lung cancer lesions, the average gross tumor volumes ( GTV ) delineated on PET and CT are ( 7.36 ± 1.62 ) ml and (8.31 ±2.05) ml, respectively (t = -1.26, P＞0.05). Conclusion The proposed threshold algorithm for tumor contouring on PET image could provide comparable GTV with CT.

Objective To observe the role of rosiglitazone in unclipped kidneys of two-kidney- one-clip hypertensive rats and examine its relationship to angiotensinⅡreceptors.Methods Two- kidney-one-clip hypertensive rats were divided randomly into 4 groups as follows:positive control group (CONT),traditional antihypertensive drugs group (TAHD,reserpine 50?g?kg~(-1)?d~(-1), dihydralazine 6.25 mg?kg~(-1)?d~(-1) and hydrochlorothiazide 6.25 mg?kg~(-1)?d~(-1),regular-dose rosiglitazone group (RRGL,rosiglitazone 5 mg?kg~(-1)?d~(-1)),and high-dose rosiglitazone group (HRGL, rosiglitazone 20 mg?kg~(-1)?d~(-1)).Sham operation rats were as negative controls.Each group had 8 rats. Animals were monitored and sacrificed at 10th week.Results Blood systolic pressure in TAHD group and HRGL group was significantly lower than that in CONT group [TAHD(137?27 ) mm Hg and HRGL (143?16) mm Hg vs CONT (191?25 ) mm Hg,P＜0.05],but no significant difference between the former two groups was found.Nor did the blood systolic pressure between RRGL group [(176?18) mm Hg] and CONT group.At 10th week,rats in SHAM group and treated groups had lower urinary urinary protein excretion rate,glomerular injury score and wall-to-lumen ratio of arteriole than those in CONT group [vs CONT urinary protein excretion rate (44.60?17.40) mg/24 h,P＜0.05; vs CONT glomerular injury score 60.85?33.05,P＜0.05;vs CONT wall-to-lumen ratio of arteriole 2.33?1.01,P＜0.01,except TAHD group].Though with the similar level of blood pressure,blood glucose and lipid,HRGL,compared with TAHD group showed lower urinary protein excretion rate [HRGL (16.78?3.50) mg/24 h vs TAHD (27.94?12.79) mg/24 h,P＜0.05],decreased glomerular injury score (HRGL 18.04?7.76 vs TAHD 27.92?6.39,P＜0.05) and wall-to-lumen ratio of arteriole (HRGL 1.75?0.38 vs TAHD 2.16?0.90,P＜0.05) in the cortexes of unclipped right kidneys.The expression of type 1 angiotensinⅡreceptor (AT1R) mRNA was no difference in HRGL group and TAHD group,but the expression of type 2 angiotensinⅡreceptor (AT2R) mRNA was more intensive in HRGL group.Conclusion Rosiglitazone can protect the kidneys from hypertensive injury,especially in high dose.The beneficial effects seem incompletely dependent on the metabolism modulating and reduction of blood pressure,but in relationship to the upregulation of AT2R mRNA.

Seven emotions generation relates to the interaction between individual and external objective matters,which is accompanied with interaction between internal desire and external matters.The activity of seven emotions bases on essence of zang-fu organs,being regulated by five zang viscera.Heart is the upstream controller,liver is the key organ to maintain normal emotionds,spleen and stomach is the hub of emotional activities,lung is the auxiliary organ, kidney is origin of emotional generation.Five zang viscera's cooperation and interaction generate the seven emotions.

Objective To investigate clinical characteristics of relapsing polychondritis(RP)and to improve early recognition for it.Methods Clinical and laboratory data of 56 patients with RP were analyzed retrospectively.Results Ratio of number of male patients to female ones was 1.2.Age at onset was(46?11)years(ranging from 27 to 71)and average interval between onset and diagnosis was(21? 35)months,(8?6),(16?31)and(29?37)months for patients initial onset with auricle,respiratory tract and joints involved,respectively.Site involved included airway in 40 patients(71.4%),auricle in 32 (57.1%),joints in 32(57.1%),eyes in 27(48.2%),nasal chondritis in 25(44.6%)and inner ear in 13(23.2%).At initial stage of the course,17 patients were misdiagnosed as respiratory infection (30.4%),nine as perichondritis(16.1%),six as pulmonary tuberculosis(10.7%),five as rheumatoid arthritis(8.9%).Seven of 40 patients with airway involvement received metallic stents for their tracheobronchial stenosis.Four patients whose condition never improved after regular therapy all had respiratory involvement.Conclusions Patients of RP with initial onset at non-auricle,non-nasal sites tended to be misdiagnosed.Prevalence of airway involvement was not so low with a poor prognosis in patients of RP.