Objective To obtain a single toxin component from the jellyfish Cyanea capillata and provide a foundation for the further study on bioactivity and function of the serine proteases from C.capillata.Methods Primers designed with restriction enzyme were used to amplify the coding region of cDNAs (CcSP1, CcSP2 and CcSP3).PCR fragments were ligated with the pET-24a( +) vector to construct the recombinant plasmids (pET24a-CcSP1, pET24a-CcSP2 and pET24a-CcSP3).After screening and identification,the recombinant plasmids were transformed into the Rosetta (DE3).plysS for protein expression.After induction with IPTG, SDS-PAGE and Western-blot were used to detect the expression of the recombinant proteins.Results SDS-PAGE showed that the proteins of rCcSP1, rCcSP2 and rCcSP3 were expressed in a single band at about 34 kDa, 42 kDa and 42kDa, respectively.Western-blot detection with anti-His antibody further confirmed that these recombinant proteins were His-tagged CcSP1, CcSP2 and CcSP3 fusion protein were obtained.Conclusion Prokaryotic recombinant plasmids of C.capillata serine proteases are contructed and recombinant proteins are obtained, which establishes the foundation for future study on the function of serine proteases from jellyfish.

Objective To investigate the inhibitory effect of melatonin on the proliferation activity of murine foregastic carcinomac ( MFC) cells via melatonin membrane receptors MT 2 and its relationship with the signaling pathways of mitogen-activated protein kinases ( MAPKs), phosphatidylinositol 3-kinase ( PI3K)-Akt.Methods Using siRNA technology to silence MT2 expression, we examined the ability of melatonin to inhibit the proliferation activity of MFC cells and its influence on the phosphorylation of ERK 1/2 and Akt.Results We found two interesting effects of SiRNA-mediated silencing of MT2 expression.Firstly, it significantly antagonized the inhibitory effect of melatonin on the proliferation activity of MFC cells .Secondly , it partially blocked the inhibitory effect of melatonin on the phosphorylation of ERK 1/2 and Akt.Conclusion Our results suggest that melatonin can inhibit the phosphorylation of ERK 1/2 and Akt via MT2 receptors , thereby inhibiting the proliferation of gastric cancer cells .

Objective To examine the effects of glucose on the expression of P38 MAPK and TGF?2 in cultured human umbilical vein endothelial cells.Methods Human umbilical vein endothelial cells(HUVECs)were incubated in a culture medium with 11.2 mmol/L,16.8 mmol/L and 33.6 mmol/L glucose concentrations for 24 h,48 h and 72 h respectively.The expression P38 MAPK and TGF?2 was measured by RT-PCR and Western-blot.Results When D-glucose concentrations rose,HUVECs exposed to high glucose concentration(11.2 mmol/L,16.8 mmol/L and 33.6 mmol/L)showd increase in cell expression of P38 MAPK and TGF?2 after 48 h and 72 h exposure as compared with those cultured in medium of low glucose concentration(5.6 mmol/L).ConclusionHigh concentration of glucose can arrest the proliferative response.Even a short-term exposure of endothelial cells(ECs)to high glucose concentration may lead to their activation associated with increased expression of P38 MAPK and TGF?2.High Glucose Concentration on P38 MAPK and TGF?2 expression may participate the pathogenesy of diabetic retinopathy.

Objective To introduce the technique and experience in reconstruction of posterior crueiate ligament-posterolateral comer(PCL-PLC)injuries with only one allograft of Achilles tendon. Methods The instable knees in 12 cases with PCL injury combined with three degree chronic PLC injury were treated with PCL reconstruction under arthroscope and PLC reconstruction through posterolateral arc incision.Single bundle grafts of PCL reconstructions in tibial and femoral tunnels were fixed by resorption screws.Fibular collateral ligament(FCL)and popliteofibular ligament(PFL)were reconstructed with reforming Larson(?)method.All reconstruction grafts only needed one Achilles tendon as donator.Total op- eration time was 130 minutes including 90 minutes of PCL reconstruction and 40 minutes of PLC recon- struction.Gradual weight loading was allowed after six weeks of bracing.Results Follow-up for mean 12 months(5-24 months)indicated that tibial“step off”reduction was 83%(10/12)and posterior drawer test of 0-1~+ 75%(9/12).Dial sign evaluated that normal external rotation angle was 75%(8/12).Nor- real varus stress test at 30?knee flexion accounted for 83%(10/12).Scores of Lysholm,Tegner and HSS were 90.5,5.1 and 84.5,respectively(P＜0.01=.Conclusion Synchronized reconstruction of PCL and PLC injuries with only one Achilles tendon can obtain satisfactory clinical result,with less expense and shorter operation time.

Objective In order to provide clinical evidence of reducing postoperative complications of procedure for prolapse and hemorrhoids and observe the effects of constipation on postoperative complications of procedure for prolapse and hemorrhoids.Methods One hundred and sixty cases of hemorrhoid were divided into two groups according to whether suffering from constipation:there were eihgty cases who were suffered from constipation in the experimental group and eighty cases who were not suffered from constipation in the control group.All patients' postoperative bleeding,anal edema,stool impaction and satisfaction were observed within 30 days and these data were compared and analysed by using SPSS 13.0 software.Results In the experimental group,the postoperative bleeding incidence was 37.5 ％ (30/80),the anal edema incidence was 31.3 ％ (25/80),the stool impaction inci dence was 8.6％ (7/80),and the patients' satisfaction rate was 88.6％ (71/80).In the control group,the postoperative bleeding incidence was 18.6％ (15/80),the anal edema's incidence was 15.0％ (12/80),the stool impaction incidence was 1.3％ (1/80),and the patients' satisfaction rate was 97.5％ (78/80).All the indexes above were different statistically in the two groups (P ＜ 0.05).Conclusion constipation could increase complications and reduce the degree of satisfaction after procedure for prolapse and hemorrhoids.

Objective To observe the changes of serum potassium concentration in rectal cancer patients from the admission to before the operation,to provide clinical evidence for preventing serum potassium disorder during and after their operation.Methods Monitoring of the changes of serum potassium concentration of 40 cases of rectal cancer patients according to three time points including the first day of admission,before taking of cathartics and the morning before operation.They were divided into different groups according to their age(young,middle age and elderly group),gender(male and female group),diet(normal,decreased slightly and serious decline group) and course(long,middle and short course group) of disease.The serum potassium concentration was observed between each time point of each group and SPSS13.0 statistical software was used to analyze the changes above.Results The mean of all patients' serum potassium concentration was (4.09 ± 0.62) mmol/L on the first day of admission,was (3.83 ± 0.46) mmoL/L before taking of cathartics that was decreased compared to the former and there was significant difference (P ＜ 0.01),it was (3.36 ±0.40) mmol/L on the morning of operation and had significant difference with the above two groups (P ＜ 0.01).It hadn't significant difference between different gender and age groups at different time points (P ＞ 0.05) except between the young group and the elderly group in the morning of the operation (P ＜ 0.05).There were statistically significant differences between the different diet condition and different course at anytime point (P ＜ 0.01).Conclusions The rectal cancer patients were possiblely in hypokalemia before operation,the causes might be relative to the long course of disease,the decreased diet,the aged and infirm,the diet structure change after admission and taking cathartics cleaning intestinal tract.

Objective To analyze the pitfalls of 18F fluorodeoxyglucose (FDG) positron emission tomography/computer tomography (PET-CT) scan in the diagnosis of 60 patients of tuberculosis mimicking malignancy.Methods The study included 60 patients with PET-CT diagnosis of probable malignancy.Fifty patients were proved to be tuberculosis by pathological examinations and 10 were diagnosed by clinical followup.The images of whole body were acquired at 60 min after administration of 222-555 MBq 18F-FDG.The PET-CT imaging characteristics and clinical data,including lesion size,distribution,standardized uptake value (SUV) were retrospectively analyzed.After the whole body scan of PET-CT,each patient had a chest spiral CT scan for detailed observation of lung lesions.Contrast enhanced CT (CECT) was performed in 8 patients.Results (1)Thirty patients were misdiagnosed as lung cancer,14 patients as malignant lymphoma,6 patients as malignant mesothelioma,3 as intestine carcinoma,2 as bone malignancy,1 patient as hepatocarcinoma,spleen malignancy,ovarian cancer,laryngocarcinoma and nasopharyngeal carcinoma respectively.(2) 90.9％ (20/22) of patients showed normal level of serum CEA and 100％ (13/13) of patients showed normal level of CA199.Increasing serum CA125 was found in all patients (6/6) with activeTB patients accompanied with ascites,pleural fluid and (or) pericardial effusion.(3) 93.3％ (28/30)active tuberculosis showed accumulated 18F-FDG which was incorrectly interpreted as malignancy.The most common sites of TB lymphadenopathy were bilateral cervical tissues,which was accounted for 85.7％(12/14).CECT revealed characteristics of peripheral enhancement and central necrosis in tubercular lymphadenopathy,which was 87.5％ (7/8).Conclusions The diverse manifestations of TB on imaging and high uptake of 18F-FDG on PET imaging result in misdiagnosis of malignancy.It is important for radiologists and nuclear medicine physicians to identify the common imaging features and patterns of TB to make a correct diagnosis.Integration of reconstruction HR CT,PET-CT and lab examinations may improve the diagnostic accuracy.

In order to investigate the role of the PTEN expression in carcinogenesis and development of endometrial carcinoma and clarify whether and how PTEN and PI3K/Akt pathway relate to endometrial carcinoma, the expression of PTEN and phospho-Akt was detected by semiquantitative reverse transcription-polymerase chain reaction (RT-PCR) methods and Western-blot from 24 cases of endometrial carcinoma, 10 cases of endometrial atypical hyperplasia, 10 cases of endometrial hyperplasia, and 10 cases of normal endometrium. SP immunohistochemical methods were used to measure levels of PTEN protein expression in following 5 study groups: 31 cases of endometrium in proliferative phase, 30 cases of endometrium in secretory phase, 71 cases of endometrial hyperplasia, 25 cases of atypical hyperplasia and 73 cases of endometrial carcinoma. Immunostaining score of PTEN was 3.39+/-0.15 in proliferative phase, 1.90+/-0.21 in secretory phase, 3.34+/-0.29 in endometrial hyperplasia, 0.62+/-0.11 in atypical hyperplasia, and 0.74+/-0.19 in endometrial carcinoma, respectively. PTEN mRNA relative value in normal endometrium, endometrial hyperplasia, endometrial atypical hyperplasia, and endometrial carcinoma was 2.45+/-0.51, 2.32+/-0.32, 0.46+/-0.11, and 0.35+/-0.13 respectively. The expression levels of PTEN mRNA and protein in patients with endometrial carcinoma and atypical hyperplasia were significantly lower than in those of proliferative phase and with endometrial hyperplasia. The level of PTEN expression in patients with endometrial carcinoma was significantly related to tissue type (P<0.005), differentiation (P<0.05) and clinical stage (P<0.05), but not to depth of myometrium invasion (P>0.05). Western blot analysis revealed that Phospho-Akt level in PTEN negative cases was significantly higher, and there was a negative correlation between PTEN and phospho-Akt (r=-0.8973, P<0.0001). It was suggested that loss of PTEN expression was an early event in endometrial tumorigenesis. The phosphorylation of Akt induced by the loss of PTEN took part in the tumorigenesis and development of endometrial carcinoma.

OBJECTIVETo compare the clinical efficacy in the treatment of post-stroke shoulder-hand syndrome between floating-needle therapy and conventional acupuncture on the basis of rehabilitation training.

METHODSOne hundred cases of post-stroke shoulder-hand syndrome were randomized into a floating-needle group and an acupuncture group, 50 cases in each one. The passive and positive rehabilitation training was adopted in the two groups. Additionally, in the floating-needle group, the floating-needle therapy was used. The needle was inserted at the site 5 to 10 cm away from myofasical trigger point (MTrP), manipulated and scattered subcutaneously, for 2 min continuously. In the acupuncture group, the conventional acupuncture was applied at Jianqian (EX-UE), Jianyu (LI 15), Jianliao (TE 14), etc. The treatment was given once every two days, 3 times a week, and 14 days of treatment were required. The shoulder hand syndrome scale (SHSS), the short form McGill pain scale (SF-MPQ) and the modified Fugl-Meyer motor function scale (FMA) were used to evaluate the damage severity, pain and motor function of the upper limbs before and after treatment in the two groups. The clinical efficacy was compared between the two groups.

RESULTSSHSS score, SF-MPQ score and FMA score were improved significantly after treatment in the two groups (all P < 0.01), and the improvements in the floating-needle group were superior to those in the acupuncture group (all P < 0.05). The total effective rate was 94.0% (47/50) in the floating-needle group, which was better than 90.0% (45/50) in the acupuncture group (P < 0.05).

CONCLUSIONThe floating-needle therapy combined with rehabilitation training achieves a satisfactory efficacy on post-stroke shoulder-hand syndrome, which is better than the combined therapy of conventional acupuncture and rehabilitation training.

Acupuncture Therapy ; instrumentation ; methods ; Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Male ; Middle Aged ; Reflex Sympathetic Dystrophy ; etiology ; rehabilitation ; therapy ; Stroke ; complications ; Treatment Outcome ; Young Adult

OBJECTIVEX-linked adrenoleukodystrophy (ALD) is a genetically determined disorder that involves the nervous system white matter, axons, adrenal cortex and testes. The typical clinical manifestations are progressive psychomotor regression, vision and/or auditory impairment and adrenal insufficiency. The clinical manifestation, biochemical change and genetic counseling work of X-linked ALD were analyzed.

METHODSThe clinical features of 29 cases with ALD were summarized and analyzed, including symptoms and signs, measurement of blood very long chain fatty acids (VLCFA), adrenal function, cranial magnetic resonance imaging (MRI) and pedigree investigation.

RESULTSAmong these 29 cases, the clinical phenotype could be classified into childhood cerebral (22 cases), adolescent cerebral (4 cases), adrenomyeloneuropathic (1 case), Addison's disease (1 case) and asymptomatic or presymptomatic (1 case) types. Nine of them had positive family history. Pedigree investigation was consistent with typical sex-linked recessive inheritance. There were 45 ALD patients in these 29 pedigrees. The neurological manifestations varied among members of the same family. Nine cases died during follow up. The causes of death were central respiratory failure or other complications of ALD and so on. Laboratory tests demonstrated abnormally high plasma levels of VLCFA in ALD patients; MRI demonstrated symmetric butterfly-like low T(1) and high T(2) signals in the parieto-occipital white matter. The impairment in the splenium of corpus callosum made the bilateral lesion region converge into one. It could progress anteriorly and injure the bilateral posterior limb of internal capsule and the temporal lobe, and could injure the brainstem inferiorly. Following intravenous injection of contrast material, thin stripe of lacelike enhancement could be observed.

CONCLUSIONSThe atypical initial symptom of ALD was seizures. The MRI showed abnormal signal in the cerebellar white matter. This disease can influence the normal development of children, this was more pronounced in the childhood cerebral ALD type. It tended to progress rapidly with dementia, vegetative state or death. Since antenatal diagnostic method is available now, emphasis should be made on the antenatal examination in order to make an early diagnosis and abort pregnancy if necessary.

Adolescent ; Adrenoleukodystrophy ; blood ; diagnosis ; therapy ; Child ; Child, Preschool ; China ; Fatty Acids ; blood ; Female ; Follow-Up Studies ; Humans ; Male ; Pedigree ; Treatment Outcome