Objective To explore the effects of light on the expression patterns of clock and clock-related genes in peripheral lymphocytes.To develop basic knowledge needed for clinical application of the circadian clock.Methods One hundred Sprague-Dawley rats were housed under constant dark(DD)or normal light-dark(LD 12:12)conditions for six weeks.Peripheral iymphoeytes were collected at different time points.The expression level of the clock gene and melatonin receptor genes mt1 and mt2 were detected using semi-quantitative RT-PCR.The data were analyzed with cosine software.Results Circadian expression of the genes was observed in both groups,but the peak phase,amplitude and strength of expression of each gene differed with the light conditions.Conclusion Light influences the expression of clock and clock-related genes in rats' peripheral lymphocytes.The clock gene might play an important role in regulating the expression of mt1 and mt2.

ObjectiveTo detect the features of dopamine secretion of cultured bovine retinal pigmentary epithelial(RPE) cells. The level of dopamine and survival rate after passage and microencapsulation were also observed. MethodsThe primary culture of bovine RPE cells was made using enzyme digestion. After purification and identification the growth curve of the cell was observed. Alginate-polylysine-alginate(APA) microencapsulated cells were made with a high voltage electostatic system. The activity of the cells in the mirocapsule was investigated by trypan blue staining. The secretion of dopamine was determined by high performance liquid chromatography (HPLC) assay. ResultsThe cell had high purity of immunocytochemistry. The growth curve showed that the exponential growth occurred at the first 1~4 days. Dopamine content was first detected at the time point of 2 hour, and arrived at the peak at about 48 hour of the cultivation. The secretion of dopamine was not different between passages. Dopamine secretion was dramatically decreased in the first 4 days after microencapsulation (P

Objective To establish a model of malignant transformation of human cells in vitro to study the lung cancer induced by radon and cigarette smoke. Methods The immortalized human bronchial epithelial cells BEAS-2B were divided into control group( C ), radon group ( Rn), cigarette smoke group (Sm) and combined group (Rn-Sm). Cells were planted onto transwell membrane one day before exposure and were directly exposed to radon and cigarette smoke pumped in a gas inhalation box. After the exposure cells were trypsinized into dishes for further growth and malignancy transformation phenotype was detected in order to compare the effects due to radon and cigarette smoke exposure. Results BEAS-2B cells showed malignantly transformed phenotype by exposure to radon and cigarette smoke. A series of sequential steps emerged among transformed cells, including altered growth kinetics, resistance to serum has changed from 0. 31 ± 0. 18 to 1.92 ± 0. 27,2. 03 ± 0. 14,2.95 ± 0. 60, and anchorage-independence growth increased from (0.01 ±0.02)% to (4.89 ±0.30)%,(8.36 ±0.50)%,(11.74 ±0.69)%.After being subculture for 20 generations, cell apoptosis of the fifth generation cells exposed to radon,cigarette smoke and both was significant decreased from ( 11.76 ± 0. 17 ) % to (4. 62 ± 0. 42 ) %、 ( 8.63 ±0. 15 )%、 (3.68 ± 0. 33 )%. Conclusions BEAS-2B cells could be malignancy transformed by radon and cigarette smokein vitro, which could be used as a cell model in lung bronchial carcinogenesis.

Objective To analyze and summarize the risk factors of failed internal fixation for intertrochanteric fracture.Methods From April 2008 to April 2011,267 patients with intertrochanteric fractures in 4 hospitals were treated with internal fixation.The relationship between the failure of internal failure and possible factors as age,gender,hypertension,diabetes,the abuse of alcohol and tobacco,use of glucocorticoid,the degree of osteoporosis and fractures type were studied.According to the surgical risk assessment table,the patients were divided into low-risk,mid-risk,and high-risk group.The rate of internal fixation failure was compared in the 3 groups.Results We found 42 cases which showed radiographic failures.The internal fixation failure directly related with advanced age,diabetes,severe osteoporosis,unstable type fracture,but not gender,hypertension,the abuse of alcohol and tobacco,use of glucocorticoid.Risk factors of internal fixation failure included diabetes,osteoporosis degree,and fracture stability.Failed intertrochanteric fracture fixation mainly occurred in the mid-risk and high-risk groups.Conclusion Severe osteoporosis,unstable fracture,diabetes are risk factors of failure of intertrochanteric fracture fixation.These factors will affect the quality of surgery.For the patient with intertrochanteric fractures in the low-risk groups,internal fixation should be the first choice for treatment.For the patients in the mid-risk and high-risk group,internal fixation should be applied cautiously.For the aged patients in high-risk groups,hip arthroplasty is a wise option.

Objective To assess the diagnostic value of 99Tcm-ECD SPECT for hyperacute cerebral ischemia using rats models.Methods A stable and permanent acute cerebral ischemia model using unilateral middle cerebral occlusion was tested in 24 healthy SD rats.The rats were randomly divided into 6 groups according to the time duration between imaging and induced-ischemia (1,2,3,4,5 and 6 h,respectively).The rats were sacrificed immediately after 99Tcm-ECD SPECT/CT imaging and then the brain tissue was dissected for triphenyl tetrazolium chloride (TTC) and HE staining.The count ratio of affected cortex to the contralateral cortex of ＜ 50％ was defined as ischemia on micro SPECT/CT.The volume of the ischemic area was calculated on both SPECT/CT and TTC images.Paired t test was used to determine the statistical difference between the volumes on SPECT/CT and TTC staining.Results The ischemia volume evaluated by TTC staining at 1,2,3,4,5 and 6 h after occlusion was (73.98 ± 27.76),(90.75 ±29.00),(135.00±40.83),(136.25±22.51),(158.50±32.72) and (168.00±32.75) mm3,respectively.The corresponding ischemia volume evaluated by micro SPECT/CT was (98.50 ± 27.77),(110.40±26.80),(157.00±36.82),(165.50±26.54),(175.75±31.16) and (177.25 ±34.33) mm3,respectively,which was concordant with that by TTC staining at each time point (t:-1.681 to-0.390,all P ＞0.05).The ischemic area on micro SPECT/CT imaging was consistent with the pink area by TTC staining.The volume evaluated by micro SPECT/CT tended to be constant 3 h after the occlusion.The ischemia volume showed no significant difference among 3,4,5 and 6 h (all P ＞ 0.05).Conclusions Micro SPECT/CT may have an haemodynamic value for evaluating in vivo cerebral ischemia applied in a rat model.It might have clinical value for the evaluation and decision-making of ultra acute cerebral infarctions.

Objective To report clinical study of diabetic foot with microsurgical treatment.Methods 32 cases basing on physical treatment underwent operation which included reconstruction of vessel under DSA and flap transfer and relaxation of nerves.Results 8 eases were examined with DSA after operation,it showed that the bypass grafts were unobstructed and the distal blood were improved;All flap were lively. Conclusion The ulcer of the patients with diabetic foot was closed early and the blood supply of the limb have been reconstructed by microsurgical treatment,it can not only avoid amputation or lower the limb amputation level,but also improve the life quality of patients and obtain social benefit.

To have an overview of the international status and trends of complementary and alternative medicine (CAM) research, and to provide a reference tool for researchers.

Objective To study the clinical significance of the mRNA expression level of a novel gene which encodes a kind of transmembrane prostate protein induced by androgen-PMEPA1, as it may predict the progress of prostate cancer from hormone-dependent to hormone-independent. Methods We used Real-time PCR to detect the mRNA expression of PMEPA1 and GSTP1 in prostate cancer cell lines (LNCaP, PC-3), epithelia cells of benign prostatic hyperplasia and tissues from 33 patients with prostate cancers and 16 cases of prostatic hyperplasia. Results We found the mRNA expression of GSTP1 and PMEPA1 were both down-regulated in prostate cancer cell lines. The mRNA expression of GSTP1 was up-regulated in 6.1% of cases, down-regulated in 81.8%, and showed no difference in 12.1%. While PMEPA1 was highly expressed in 27.3% of cases, lowly expressed in 27.3%, and not differently expressed in 45.4%. Statistical analysis showed that the mRNA expression of GSTP1 was relevant to ages, but had no relationship with PSA, TNM stage, osseous metastasis or tumor differentiation, while the mRNA expression of PMEPA1 was relevant to osseous metastasis and tumor differentiation, but had no relationship with age, PSA or TNM stage. Conclusions PMEPA1 is possibly a useful biomarker, as it can identify patients with unfavourable prognosis, however, this hypothesis needs to be further studied with large samples.

The aim of this study was to observe and compare the endogenous circadian rhythm and photoresponse of Clock gene transcription in the suprachiasmatic nucleus (SCN) and pineal gland (PG) of rats. With free access to food and water in special darkrooms, Sprague-Dawley rats were housed under the light regime of constant darkness (DD) for 8 weeks (n=36) or 12 hour-light: 12 hour-dark cycle (LD) for 4 weeks (n=36), respectively. Then, their SCN and PG were dissected out every 4 h in a circadian day, 6 rats at each time (n=6). All animal treatments and sampling during the dark phases were conducted under red dim light (<0.1 lux). The total RNA was extracted from each sample and the semi-quantitative RT-PCR was used to determine the temporal mRNA changes of Clock gene in the SCN and PG at different circadian times (CT) or zeitgeber times (ZT). The grayness ratio of Clock/H3.3 bands was served as the relative estimation of Clock gene expression. The experimental data were analyzed by the Cosine method and the Clock Lab software to fit original results measured at 6 time points and to simulate a circadian rhythmic curve which was then examined for statistical difference by the amplitude F test. The main results are as follows: (1) The mRNA levels of Clock gene in the SCN under DD regime displayed the circadian oscillation (P<0.05). The endogenous rhythmic profiles of Clock gene transcription in the PG were similar to those in the SCN (P>0.05) throughout the day with the peak at the subjective night (CT15 in the SCN or CT18 in the PG) and the trough during the subjective day (CT3 in the SCN or CT6 in the PG). (2) Clock gene transcription in the SCN under LD cycle also showed the circadian oscillation (P<0.05), and the rhythmic profile was anti-phasic to that under DD condition (P<0.05). The amplitude and the mRNA level at the peak of Clock gene transcription in the SCN under LD were significantly increased compared with that under DD (P<0.05), while the value of corresponding rhythmic parameters in the PG under LD were remarkably decreased (P<0.05). (3) Under LD cycle, the circadian profiles of Clock gene transcription induced by light in the PG were quite different from those in the SCN (P<0.05). Their Clock transcription rhythms were anti-phasic, i.e., showing peaks at the light phase ZT10 in the SCN or at the dark time ZT17 in the PG and troughs during the dark time ZT22 in the SCN or during the light phase ZT5 in the PG. The findings of the present study indicate a synchronous endogenous nature of the Clock gene circadian transcriptions in the SCN and PG, and different roles of light regime in modulating the circadian transcriptions of Clock gene in these two central nuclei.
Animals ; CLOCK Proteins ; genetics ; Circadian Rhythm ; physiology ; Male ; Photoreceptor Cells, Vertebrate ; physiology ; Pineal Gland ; physiology ; Rats ; Rats, Sprague-Dawley ; Suprachiasmatic Nucleus ; physiology ; Transcription, Genetic

OBJECTIVETo study the effect of Bushen Wenyang Huayu Recipe (BSWYHYR) on nerve growth factor (NGF) and transient receptor potential vanilloid receptor I (TRPV1) in experimental endometriosis (EMT), and to explore its mechanism for treating EMT-induced pain.

METHODSTotally in-bred line BALB/c 75 female mice were divided into five groups, i.e., the sham-operation group, the model group, the high dose BSWYHYR group, the low dose BSWYHYR group, the gestrinone group, 15 in each group. Writhing response was observed in each group. Serum contents of NGF were detected using ELISA. Expression levels of NGF and TRPV1 in uterus and ectopic foci were detected using immunohistochemical staining SP and Western blot. mRNA expression levels of NGF and TRPV1 in uterus and ectopic foci were detected by Real-time PCR.

RESULTSThe serum NGF content in the model group was higher than that in the sham-operation group (P < 0.01), and there was positive correlation between NGF and the writhing frequency (r = 0.574, P < 0.01). Compared with the model group, serum levels of NGF significantly decreased in the 3 treatment groups (P < 0.05). Compared with the sham-operation group, mRNA and protein expression levels of NGF and TRPV1 increased significantly in the model group (P < 0.01). Protein expression levels of NGF and TRPV1 decreased significantly in the 3 treatment groups, when compared with the model group (P < 0.01). mRNA expression levels of NGF and TRPV1 decreased most in the high dose BSWYHYR group (P < 0.01). NGF in uterus and ectopic foci was positively correlated with protein and mRNA expression levels of TRPV1 (P < 0.01).

CONCLUSIONSNGF and TRPV1 participated in the occurrence of pain in EMS. BSWYHYR played an important role in inhibiting EMT-induced pain through reducing the up-regulation of NGF on TRPV1.

Animals ; Drugs, Chinese Herbal ; therapeutic use ; Endometriosis ; Female ; Humans ; Mice ; Mice, Inbred BALB C ; Nerve Growth Factor ; metabolism ; Pain ; RNA, Messenger ; TRPV Cation Channels ; metabolism ; Up-Regulation ; Uterus