AIM To establish an HPLC method for the simuhaneous content determination of five flavonoids in Duchesnea indica (Andr.) Focke at five picking time (April,May,June,July and August).METHODS The analysis of D.indica methanol extract was performed on a 25 ℃ thermostatic Agilent ZORBAX SB-C18 column (250 mm×4.6 mm,5 μm),the mobile phase comprising of acetointrile-0.1％ methanoic acid flowing at 1.0 mL/min in a gradient elution manner,and the detection wavelength was set at 350 nm.RESULTS Rutin,hyperoside,isoquercitrin,celereoin and kaempferol showed good linear relationships within their own ranges (R2 ≥0.998 6),whose average recoveries were 97.1％-101.5％ with the RSDs of 1.37％-2.37％.The contents of five constituents in samples at different picking time exhibited obvious differences,among which lutin and hyperoside contents were the highest in June,isoquercitrin content was the highest in July,and celereoin and kaempferol contents were the highest in August.CONCLUSION The suitable picking time of D.indica is June and July.

Objective To investigate the effect of BML-111 (the analogue of lipoxin) on uterine Hela cell (cervix cancer cell line) proliferation and the underlying mechanism. Methods Hela cells were stimulated by 50, 100, 200 and 400 μg/L BML-111, respectively, and cell viability was determined by MTT assay. Hela cells were divided into three groups:the control group (no treatment), the BML-111(200μg/L) group and the BML-111(200μg/L)plus Boc-2 (10μmol/L)group. Expression and location of P53 protein were detected by immunofluorescence. Expressions of NF-κB p65,P53 and CyclinD1 protein were detected by Western blotting. Results BML-111 (100, 200 and 400 μg/L) could effectively inhibit Hela cell viability compared with the control group (P < 0.05). P53 expression was shown decreased in both the nucleus and the cytoplasm without any change of P53 location , however, Boc-2 could reverse this effect. BML-111 could effectively inhibit P53 and CyclinD1 expression via NF-κB pathway and the effects could also be inhibited by Boc-2. Conclusions BML-111 can effectively inhibit Hela cell proliferation via FPR2 and NF-κB pathway.

AIM To observe the antinociceptive effects of glucosides of Chaenomeles speciosa(GCS) and to study their relative mechanism. METHODS The effects of GCS on normal and inflammatory animals were observed in mouse acetic acid writhing test, mouse formalin test and arthritic flexion test of adjuvant arthritis(AA) rats; the concentration of prostaglandin E2 (PGE2) and tumor necrosis factor-α(TNF-α) secreted by the synovial cells of AA rats were measured by radioimmunoassay. RESULTSDifferent doses of GCS (60, 120, 240 mg·kg-1 for mice and 30, 60, 120 mg·kg-1 for rats, ig) inhibited mice′s writhing response and second phase of formalin response. It also suppressed the increased arthritic flexion scores in AA rats. On 28 d after inflammation induction, GCS (60, 120 mg·kg-1) decreased the concentration of PGE2 and TNF-α of synovial cells in the AA rats. CONCLUSIONGCS have antinociceptive effects, which related to its inhibitory effects on peripheral inflammatory mediators.

ce, sIMRT and IMRT radiotherapy techniques can protect the lung and spinal cord well.

Objective To explore the clinical features, treatment and prognosis of intrabepatic biliary cystadenocarcinoma (IBC). Methods We retrospectively analyzed clinical data of 17 patients with pathologically confirmed IBC, treated between January 2002 and September 2007. Results Of the 17 patients, 4 were men and 13 were women, with a mean age of 49.3 years (range 45 -68). Serum level of AFP was normal in all patients. CA19-9 was abnormally elevated in only two patients and the other two had abnormal serum level of CEA. Seven cases underwent radical resection, 7 underwent complete tumor resection combined with removal of tumor thrombi in the bile duct, 3 received palliative surgery. On histopathological examination, 10 were biliary cystadenocarcinoma,2 were biliary cystadenoma with partial eanceration,2 were papillary cystadenocarcinoma,2 were mutinous papillary cystadenocarcinoma and 1 was of mixed cystadenocarcinoma and hepatocellular carcinoma. According to tumor differentiation, the number of well, moderate and poor-differentiated group was 7, 3 and 7 cases respectively. The medality of surgery (β= - 0.692, P = 0.01) and tumor differentiation (β = - 2.041, P = 0.007) effected the prognosis significantly. Conclusions IBC occurs mainly in elderly women. CA19-9 examination does not help in the establishment of diagnosis of IBC. The occurrence of tumor thrombosis in common bile duct doesn't necessarily indicate poor prognosis, hence a IBC patient will still have a satisfactory prognosis should the primary tumor be completely resected and tumor thrombi removed.

Objective To investigate the diagnositic value of uhrasonography in fetuses with persistent right umbilical vein.Methods Three cases of persistent right umbilical vein diagnosed in our hospital were analyzed,including the sonographic findings and clinical outcome.Results In all cases the fetal gallbladder was located between the intraabdominal umbilical vein and the stomach,the umbilical vein was connected to the right portal vein,and the portal vein curved toward the stomach.Other intracranial malformations were detected in one case,and the baby died after birth.No addional malformations were found in the other two babies,and both of them were healthy at the age of 6 months.Conclusions Persistent right umbilical vein has typical prenatal sonographic appearances.The neonatal prognosis is favorable when other anomalies are ruled out.It's clinically important for early diagnosis and genetics consultation.

Adult ; Bronchoalveolar Lavage ; methods ; Humans ; Male ; Middle Aged ; Pneumoconiosis ; therapy ; Treatment Outcome

Carcinoma, Papillary ; genetics ; Humans ; Mutation ; Proto-Oncogene Proteins B-raf ; genetics ; Thyroid Neoplasms ; genetics

Background The retinal degeneration 11 (rd11) mouse is a newly discovered naturally occurring recessive animal model with lysophosphatidylcholine acyltransferase 1 (Lpcatl) mutation.Previous studies showed that the photoreceptor cells are characterized by typical rod-cone degeneration pattern in rd1 1 mice,while cone degeneration pattern in rd11 mice is unclcar.Objective Using immunofluorescence staining techniques with retinal wholemount,we aim to clarify the degeneration patterns of cone-function related M-opsin or S-opsin in different ages of rd1 1 mice.Methods A total of thirty rd1 1 and C57BL/6J mice at postnatal (P) day 14,28,42 (five in each age group) were sacrificed and retinal wholemounts were prepared.Immunohistochemistry was performed to identify the expression of M-opsin or S-opsin in retinal wholemounts,which were photographed with a fluorescent microscope.Cone opsins were compared between rd1 1 retinas and age-matched normal C57BL/6J retinas by manually counting the opsin positive cone cells in different quadrants of the retinas.Results The number of M-opsin or S-opsin positive fluorescent dots in each quadrant was similar at all ages of normal C57BL/6J retina.M-opsin positive fluorescent dots in dorsal/temporal,ventral/temporal,dorsal/nasal and ventral/nasal quadrants of rdl 1 retina at P28 were (414±32),(300± 8),(324 ± 22) and (250± 20)/0.037 mm2,which were lower than the age-matched normal C57BL/6J mice (t =4.114,15.225,7.505,17.990,all at P＜0.05).At the same time the S-opsin positive fluorescent dots in P28 rd11 were (8 ±4),(175 ± 16),(74 ± 13) and (315 ±20)/0.037 mm2,with significant decrease in comparison with those in the age-matched normal C57BL/6J mice (t =8.555,17.076,21.637,13.498,all at P＜0.05).With the development of retinal degeneration in rd11 mice,the M-opsin degeneration spread from central to ventral,nasal and then to temporal and dorsal peripheral retina;and the S-opsin loss started from dorsal/temporal to ventral/nasal retina.Conclusions Most of the M-opsin and S-opsins,especially the S-opsins in rd11 mice,degenerate in 6 weeks.Retinal wholemount and cone opsin immunofluorescent staining provide a useful tool to show the cone degeneration pattern and to evaluate the therapeutic efficiency in ongoing gene therapy study.

Background Trans-corneally subretinal injection in rodent model is a useful method for genetic therapy,stem cell transplantation and the study on the ophthalmic research.Standarized operation process is critical for the successful treatment.However,there is no literature to report the detailed procedure and the influence of this technique on morphology and function of retina.Objective This sudy was to introduce a method of trans-corneally subretinal injection and evaluate its influence on the morphology and function of retina.Methods Trans-corneallly subretinal injection was performed on the left eyes of 2-month-old SPF C57BL/6J mice after dilation of pupils.A 301/2G disposable needle was used to puncture the cornea within the pupil area near limbus and avoid touching the lens and irises under eye surgery microscope.Then,a 33G blunt needle was used to insert into the vitreous and toward subretinal space via corneal puncture.Normal saline with 0.1％ fluorescein sodium of 1 μl was slowly injected into the space,and 2.5％ hydroxypropyl methylcellulose was dropped on ocular surface for the observation of the fundus clearly.According to the percentage of the retina filled with subretinally injected solution,the experimental eyes were divided into 80％-100％ area group,50％-70％ area group after injection,and the mice in the pseudo-injected group,in which injection procedure stopped just before the solution was pushed in to the subretinal space did not inject any solution after punctured.The right uninjected eyes of the mice served as normal control group.Four eyes were selected for each group.The structural changes were evaluated by optical coherance tomograpby (OCT) 1 day,2 days,3 days and 5 weeks after injection,and retinal function was assessed by the recored of electroretinography (ERG) 5 weeks after injection.The retinal sepcimens were prepared to examin the morphological changes by hematoxylin and esosin staning.The use of care followd the Regulations for the Administration of Affair Concerning Experimental Animals of Zhejiang Province.Results About 70％ of the injected eyes showed that retinal blebs filled with injected green fluorescein solution occupied 50％ or more retinal area with minimal damages.The focal detachment between neurosensory retinal layer and retinal pigment epithelium (RPE) was exhibited 1 day postinjection,and almost all the retinas retached 2 days after injection.In the fifth week after injection,the amplitudes of ERG b wave were (386.25±37.88),(357.50±41.03),(324.25±53.45) and (410.50±14.88) μV in the sham operation group,50％-70％ area group,80％-100％ area group and normal control group,respectively,showing a significant difference among the 4 groups (F=3.574,P=0.047),and the amplitudes of b wave in the normal control group were higher than those in the 80％-100％ area group (all at P ＜ 0.05).The detachment between retinal neuroepithelium layer and RPE layer,cell proliferation and transposition in the outer nuclear layer were dispalyed under the light microscope in the sham operation group,50％-70％ area group and 80％-100％ area group,and the disordered outer segment of photoreceptors at the injecting area was seen in the 50％-70％ area and 80％-100％ area groups at five weeks after injection.However,retinal sructure and morphology were normal at the non-injection area.Conclusions Trans-corneally subretinal injection is an effective and safe way for subretinal injection.