Objective To study the ER status in Adriamycin-sensitive and Adriamycin-resistant MCF-7 human breast cancer cell lines. MethodsThe status of ER of MCF-7/ADR and parental MCF-7 cells was detected by Western blot. The expression of ER mRNA was detected by RT-PCR .The growth and the sensitivity to Estrogen(E 2) and droloxifene(Dro) of cells were investigated by MTT assay, and the distribution of cell cycle was detected by flow cytometric assay. Results ER and ER mRNA were positive in MCF-7 cells, and negative in MCF-7/ADR cells. In comparison with MCF-7 cells, MCF-7/ADR cells showed lower growth rate, and the cell cycle was arrested at G 0/G 1 phase. E 2 at concentrations between 1?10 -12mol/L to 1?10 -7mol/L significantly stimulated the growth of MCF-7, but did not stimulate the growth of MCF-7/ADR.Dro at concentrations between 10*!?mol/L to 20*!?mol/L significantly inhabited the growth of MCF-7, and the inhibition was dose-dependent. Dro at concentrations below 20*!?mol/L did not inhibit the growth of MCF-7/ADR, dro inhabited the growth of MCF-7/ADR only at the concentration of 20*!?mol/L, and the inhibition was more effective than MCF-7. Conclusions ER was lost in MCF-7/ADR cells,probably at mRNA level. Compared with MCF-7, the growth rate of MCF-7/ADR decreased,MCF-7/ADR cells lost the dependence on estrogen and the sensitivity to endocrine therapy.

Objective To investigate diagnosis and treatment of venous cerebral infarction.Methods The reasons, clinical findings, imaging characters, treatment methods and prognosis of 19 patients diagnosed venous cerebral infarction were analyzed.Results The most frequent symptoms were headache, focal deficits, epilepsy, increased CSF pressure.Among 19 cases, multiple cerebral lesions were found in 12 cases, lobes lesions in18 cases, cerebral infarction with hemorrhage in 3 cases. After various treatments, 16 patients were discharged and one patient died and 2 patients gave up treatment. Conclusions Venous cerebral infarction has complicated causes, non-specific chinical manifestations and characteristic MR images. Anticoagulation and thrombolysis treatment in early stage may have good therapeutic effect.

The concept of epigenetics was first proposed by Waddington in 1942,referring to phenotypic changes that are not related to the underlying DNA sequence,yet alterations in the level and function of gene expression can be heritable,such as DNA methylation,histone modifications,microRNA interference,etc.Phenotype changes while genotype remains unchanged,this controlled mechanism can be kept stable throughout cell division,proliferation,and subsequent process of development.This process thus may cause corresponding changes in the pathophysiology,which are correlated with the occurrence of cancer,immune disorder,cardiovascular disease,metabolic syndrome and so on.Diabetes mellitus is the third heaviest chronic diseases in the world,and does serious harm to human health,especially the diabetic vascular diseases.Even if the blood glucose level is controlled in an ideal state,vascular inflammation still persists,indicating the existence of metabolic memory.Studies have suggested that the pathogenesis can be elucidated at the level of epigenetics.This paper mainly focuses on DNA methylation,histone modifications,and is an overview of epigenetics research progress in diabetic vascular diseases.

A silver needle-knife which has the dual function of silver needle and needle-knife is designed. The main components of this silver needle-knife are approximately 50% silver and approximately 50% nichrome. The silver needle-knife is composed of five parts, including needle-knife tail, spiral handle; steering handle, needle-knife body and needle-knife edge. It converges the advantages of needle-knife and silver needle, which can cut loose of diseased tissue and peel adhesion of lesions, but also be heated with moxa cone and thermal therapeutic instrument, and connect with electroacupuncture apparatus. It has the function of warming channel and removing coldness, dispelling wind and eliminating dampness, resolving spasm and relieving pain, dredging the channel and so on. Due to the spiral handle and the steering handle, the operation is easier, which reduces the blindness of cutting and increase the safety. It is mainly used for soft tissue injury, rheumatism and rheumatoid arthritis, as well as degenerative diseases of spine and joint, and it has obvious efficacy on some internal medical diseases.
Acupuncture Therapy ; instrumentation ; Equipment Design ; Humans ; Needles ; standards

Objective:To analyze the stress distribution in the fixed denture of edentulous maxilla supported by 6 implants at different implant position.Methods:The skull of a volunteer with total anodontia and moderate alveolar bone absorption was scanned by CBCT.Computer software,Abaqus 6.9,Geomagic Studio 12,and Mimics10.01,were applied for data processing and reconstruction of a 3D finite element model of implant-supported fixed denture with 6 implants in different location;and then,the stress distribution was analisized.Results:3D finite element model of edentulous maxillary implant-supported fixed bridge with good geometric similarity was established.The implant neck bone interface stress value of the distal of the implant fixed bridge at the molar area,the stress concentration area,was the maximum.The main stresses were compressive in the cortical bone adjacent to the mesial or the distal implants.The shift of the denture decreased from anterior to posterior.In model Ⅰ (implants at 13,15,17,23,25 and 27) distal cantilever was not used and the stress distribution was the evenest.Conclusion:The implant position at 13,15,17,23,25 and 27 of edentulous maxillary 6-implants-supported fixed bridge was the best for the stress distribution among the 8 models.

Objective To explore the effect of different doses of dobutamine on acute lung injury (ALI) in rabbits with septic shock and to clarify the possible mechanism.Methods The rabbits model of septic shock was made by cecal ligation and puncture combined with intravenous injection of endotoxin,70 male New Zealand white rabbits were randomly divided into five groups (14 rabbits in each groups):shamc operation group (group A),ALI group (group B),dobutamine low-dose group (group C),dobutamine medium-dose group (group D) and dobutamine high-dose group (group E),7 rabbits from each group were sacrificed 3 h and 6 h after septic shock.The level of cyclic adenosine monophosphate (cAMP) in lung tissue was detected by ELISA.The expression of aquaporin 5 (AQP5) protein was determined by western blotting.The wet to dry weight (W/D) ratio was measured.The pathological and ultrastructural changes of lung tissue were evaluated by optical microscopy and electron microscope,and lung injury score was assessed.The differences among the different groups were analyzed by one-way ANOVA (LSD test).Results The level of cAMP and expression of AQP5 protein in lung tissue at 3 h and 6 h were dramatically lower in group B than those in group A (3.53 ±0.43) pmol/mLvs.(21.18 ±0.62) pmol/mL; (0.44 ± 0.04) pmol/mLvs.(0.99±0.06)pmol/mL; (2.71±0.56)pmol/mLvs.(21.78±0.62)pmol/mL; (0.29 ±0.05) pmol/mLvs.(0.91 ±0.06) pmol/mL; all P ＜0.001,while the W/D ratio was obviously higher in group B than those in group A (all P ＜0.001).Compared with group B,the level of cAMP and AQP5 protein expression in lung tissue were significantly increased at 6 h in group C (8.48 ±0.61) pmol./ mLvs.(2.71±0.56) pmol/mL,P＜0.01; (0.49 ±0.04) pmol/mLvs.(0.29 ±0.05) pmol/mL,P=0.001 and at3 hand6 hin groupDandE (10.86±0.66) pmol/mLvs.(3.53±0.43) pmol/mL; (0.60±0.05) pmol/mLvs.(0.44±0.04) pmol/mL; (13.80±0.49) pmol/mLvs.(2.71±0.56) pmol/mL; (0.64 ± 0.03) pmol/mLvs.(0.29 ± 0.05) pmol/mL; (15.57 ± 0.60) pmol/mL vs.(3.53±0.43) pmol/mL; (0.91 ±0.05) pmol/mLvs.(0.44 ±0.04) pmol/mL; (19.30±0.42) pmol/mL vs.(2.71 ±0.56) pmol/mL; (0.89 ±0.08) pmol/mL vs.(0.29 ±0.05) pmol/mL; all P ＜ 0.01,while the W/D ratio in group E was decreased obviously (P =0.002; P =0.001).Compared with group C and D,the level of cAMP and the expression of AQP5 protein at 3 h and 6 h in group E increased significantly (all P ＜0.01.The pathological and ultrastructural changes of lung tissue were more intensive in group B than those in group A and the lung injury scores were obviously higher (P ＜0.01).The degree of lung pathological and ultrastructural lesion was ameliorated after administration of dobutanmine.Additionally,histological scores decreased significantly (P ＜ 0.01).Conclusions Our study demonstrated that dobutamine could improve ALI induced by endotoxin,the mechanism of protective effect may involve in increasing the level of cAMP and up-regulating the AQP5 protein expression,and high-dose dobutamine had better effects.

OBJECTIVE:To establish the method for the simultaneous determination of clopidogrel (CLO) and its active metabolites (CATM) and inactive metabolites (CCAM),and to conduct pharmacokinetic study. METHODS:The plasma sam-ple had been derivatized by 2-bromine-3′-methoxy acetophenone(MPB),and was precipitated by acetonitrile. Using carbam-azepine as internal standard,UPLC-MS/MS was adopted. The separation was performed on Waters ACQUITY UPLC HSS T3 column with mobile phase consisted of water(containing 0.1% formic acid)-acetonitrile(containing 0.1% formic acid)using a gradient elution program at the flow rate of 0.50 ml/min. The ESI was equipped and quantitative analysis was operated in posi-tive ion and MRM mode. The mass transition ion-pairs were followed as m/z 322.1→211.8(CLO),m/z 504.1→155.0(the alkyl-ation derivatives of CATM,CATMD),m/z 308.3→198.0(CCAM),m/z 273.2→194.3(internal standard). RESULTS:The lin-ear calibration curves for CLO,CATMD and CCAM were obtained in the concentration range of 0.03-20.00 ng/ml,0.30-200.00 ng/ml and 10.00-10 000.00 ng/ml in plasma,respectively;intra-day and inter-day RSD for them were all less than 15%,and relative error(RE)ranged from -3.5% to 5.7%. Main pharmacokinetic parameters of CLO,CATMD and CCAM in 5 healthy volunteers after oral administration of CLO 300 mg were as follows:cmax were(7.89±5.46),(15.58±8.08),(8 023.33± 1 047.39)ng/ml;tmax were(1.25 ± 0.43),(1.25 ± 0.43),(1.67 ± 0.29)h;t1/2 were (2.31 ± 0.61),(0.64 ± 0.08),(6.53 ± 2.55)h;AUC0-t were(17.19±14.59),(21.39±9.58),(30 648.85±8 026.63)ng·h/ml. CONCLUSIONS:The established method is sensi-tive,rapid and convenient,which is suitable for pharmacokinetic study and plasma concentration determination of CLO and its metabolites.

BACKGROUND:There is a high risk for the elderly cervical spine surgery combined with cerebral infarction, whereas conservative treatment is often unable to resolve serious cervical lesions.
OBJECTIVE:To discuss the surgical effects of anchor fixation via posterior approach on cervical spinal canal stenosis combined with cerebral infarction.
METHODS:A total of 21 patients with cervical spinal canal stenosis combined with cerebral infarction who were admitted over the past 5 years accepted cervical posterior expensive open-door laminoplasty and fixation with wire anchors. Therapeutic effects were evaluated according to the Japanese Orthopaedic Association (JOA) scores.
RESULTS AND CONCLUSION:Al the patients were fol owed up 6 to 24 months, averagely 15 months. Al patients were smoothly through the perioperative period. There were no acute severe cerebral infarction cases. Preoperative JOA score was (7.6±2.0) points averagely, and postoperative JOA score was (13.3±1.8) points averagely, showing a significant difference (P<0.01). Of the 21 patients, excellent effects were in 11 cases and good in 7 cases. The effective rate was 95%(20/21), and the excellent-good rate was 86%(18/21). The risk of surgical treatment of patients with cerebral infarction is higher, but it is not the absolute contraindication. Choosing the proper operation way is highly important. Cervical posterior expensive open-door laminoplasty and fixation with wire anchors can be used to reduce the risk of cerebrovascular accidents to some extent, and obtain an excellent clinical effect.

Objective To report three cases of localized primary sclerosing cholangitis (PSC) mimicking a hilar cholangiocarcinoma (Klatskin tumor) and to summarize their clinical characteristics and the ways to differentiate them through a literature review.Method The clinical data of three patients with localized PSC mimicking a hilar cholangiocarcinoma were retrospectively analyzed.The characteristics of laboratory tests and imaging examination were reviewed,and therapy and prognosis were discussed.Results The three patients were all diagnosed to have a hilar cholangiocarcinoma preoperatively,but the diagnosis of PSC was confirmed by histopathology post-operatively.All the three patients had elevated CA19-9,2 patients had elevated anti-nuclear antibody (ANA) and 2 patients had elevated IgG.All the three patients underwent surgical resection and histopathological study showed chronic inflammation of the hilar bile ducts and cholangitis of the intrahepatic portal area.The three patients were followed up from 7 months to 8 years with no symptoms.Conclusions Localized PSC is rare and it can casily be misdiagnosed as a hilar cholangiocarcinoma.Biopsy before surgery is helpful for the differential diagnosis but it is difficult to get a good biopsy sample.Surgical resection is an effective treatment.

Objective To investigate the effects of heat-killed Staphylococcus aureus (HKSA) on the apoptosis and expression of iNOS and IL-1β in THP-1 monocytes in the presence of high concentration of glucose.Methods THP-1 cells were cultured in medium containing 25.0 mmol/L(HG) or 5.5 mmol/L (LG,control) D-glucose for 12 h-8 d.The THP-1 cells cultured for 6 d were extracted on the 0-48 h with or without HKSA,then apoptosis and expression of iNOS and IL-1β were examined.Apoptosis was analyzed by flow cytometry and expressions of IL-1β and iNOS were quantitated by real-time PCR.Results The expression of iNOS and IL-1β in THP-1 monocytes was increased significantly in the presence of high concentration of glucose for 12-48 h(P＜0.05),reaching the highest level at 24 h and returned to baseline after 4 d.The expression was significantly lower than that of control after 4-6 d.Apoptosis rate was also increased significantly after 48 h to 4 days.HKSA infection enhanced apoptosis,but inhibited the expression of iNOS and IL-1 β in the presence of high concentration of glucose.The expression of iNOS and IL-1β increased significantly at 6 h(P＜0.01),reaching the highest level at 12 h,but the levels were significantly lower than those in control groups (P＜0.05).Conclusion These data suggest that high concentration of glucose can interfere with the anti-bacterial function of monocytes by reducing their expression of iNOS and IL-1β and enhancing their apoptosis.