Objective] To evaluate the navel therapy of ancient formula for ehloasma of liver qi stagnation syndrome. [Method] The 40 cases take navel therapy of ancient formula, change every 24 hours for one month; make therapeutic effect observation before and after the treatment . [Result] 21 cases were cured, 10 had marked effect,7 had effective,2 had no effect, and the total effective rate was 95%.[Conclusion] Navel therapy of ncient formula has good cure effect on ehloasma.

Objective To investigate the expression and distribution of parathyroid hormone (PTH) in renal tissues of early stage chronic kidney disease (CKD),and to elucidate its potential role in renal lesion.Methods Eighty-two patients of early stage CKD (stage 1 and 2) diagnosed as glomerulonephritis (GN) with different pathologic types by renal biopsy in our department between 2009 and 2012 were enrolled in the study.Renal tissues of eight patients with mismatched HLA haplotype or the normal part of renal cancer were chosen as controls.Scr,BUN,serum calcium,phosphorus,PTH and 25(OH)VitD3 were measured.Creatinine clearance (Ccr) was calculated by Cockcroft-Gault (CG)formula.99mTc-DTPA clearance rate was used to detect GFR.Patients were divided into mild,moderate and severe groups according to the renal interstitial extent of inflammatory cells infiltration.Immunohistochemistry was used to observe the expression and distribution of PTH in renal tissues.Image-Pro Plus software was used to calculate A value of PTH in renal tissues and compare the extent of PTH expression.Results The levels of calcium,phosphorus,25(OH)VitD3 and PTH in peripheral blood from GN patients of CKD stage Ⅰ and 2 were normal.PTH had no correlation with the above indexes.PTH expression could be seen in renal tissues of all the GN patients with different pathologic types,and it mainly located in renal tubular,only a few in glomeruli and interstitium.The expression of PTH in renal tissues of GN increased compared with the controls (P ＜ 0.01).Furthermore,PTH expression elevated with the increase of inflammatory cells infiltration in interstitium.However the expression of PTH was not significantly different among different pathologic types of GN.Conclusions In the early stage CKD,PTH expression in patients of GN increases,which occurs earlier as compared to PTH elevation in peripheral blood and the imbalance of minerals and bone metabolism.The intensity of PTH expression is associated with the local inflammation.

Objective To investigate the changes and prognostic significance of bone marrow(BM) oil drop and megakaryocyte counts after chemotherapy in acute myeloid leukemia (AML) patients (non-M3).Methods Ninty-nine adult patients with denovo AML (non-M3) were retrospectively analyzed to evaluate the change of BM oil drop and megakaryocyte counts and their influences on overall survival(OS) and disease free survival (DFS) during all stages of standardized therapy.Results The median DFS and OS were 21 (2-88);months and 70 (4-89) months,respectively; and 3-year predicted DFS and OS were 47.3 ％ and 55.8 ％,respectively.After AML patients (non-M3) achieving complete remission (CR) by induction therapy,BM oil drop tended to increase along with postremission chemotherapy cycle accumulation, while megakaryocyte counts tended to decrease.The univariate analysis indicated that megakaryocyte counts decreased after the second course of postremission therapy. BM oil drop increased after the first to the third course of postremission therapy.Grade of myelofibrosis in BM biopsy,serum lactate dehydrogenase (LDH) level at diagnosis,flow cytometric immunophenotyping, the percentage of BM blast cells at diagnosis and the percentage of residual leukemic cells (RLC) during aplasia (7-10 days after the end of induction therapy) had prognostic significance.Multivariable COX analysis indicated the percentage of BM blast cells at diagnosis and change of BM oil drop after the third postremission therapy were independent prognostic factors for DFS (P =0.010,0.018 respectively),and RLCs during aplasia and change rate of the megakaryocyte counts after the second postremission therapy were independent prognostic factors for OS (P =0.009, 0.038respectively).Conclusion After AML patients (non-M3) achieving CR by induction therapy,BM oil drop tends to increase along with postremission chemotherapy cycles accumulation,while the megakaryocyte counts tend to decrease.Dynamic observations of bone marrow oil drop and megakaryocyte counts are helpful for assessing the prognosis of acute myeloid leukemia (non-M3).

Objective To explore the value of 1H-MRS on the evaluation of Alzheimer's disease (AD) with neural stem cells (NSCs) transplantation in an APP-PS1 double transgenic (tg) AD mouse model.Methods NSCs from C57BL/6 mice were cultured and amplified.APP-PS1 tg mice (n =30) aged 12 months were used as the study group,and mild-type mice (n =15) were used as the control group.Animals in the study group were randomized into two subgroups,the AD mice in one subgroup received NSCs transplantation (NSCs group) and in another subgroup received phosphate buffer saline (PBS,PBS group)in bilateral hippocampal CA1.Animals in the control group were not treated.Using a 7.0 T high-fieldstrength MR imager,1H-MRS was performed before and 6 weeks after transplantation to measure the area under the peak of n-acetyl aspartate (NAA),glutamate (Glu),myo-inositol ( mI),choline (Cho) and creatine (Cr) in the hippocampal area,NAA/Cr,Glu/Cr,mI/Cr and Cho/Cr ratio were calculated and compared with histopathological results (including Nissl's staining and electron microscope examination).Comparisons among NSCs,PBS and control groups were conducted by one-way ANOVA.Results NSCs from C57BL/6 mice were cultured successfully. Before transplantation,the mean NAA/Cr,Glu/Cr and mI/Cr in NSCs,PBS and control groups were 0.89 ± 0.05,0.88 ± 0.04 and 1.15 ± 0.05,0.40 ± 0.03,0.39 ± 0.03 and 0.45 ± 0.05,0.67 ± 0.05,0.67 ± 0.05 and 0.52 ± 0.04,respectively,and differences were statistically significant (F =148.918,7.529,59.468,P ＜ 0.01 ). There were no significant differences in NAA/Cr,mI/Cr and Glu/Cr ratios between NSCs and PBS groups before transplantation (t =0.147,0.096,0.207,P ＞ 0.05 ),but the differences were significant compared with the control group (t =0.255,0.467,0.171 and t =0.269,0.527,0.151,P ＜0.05).Six weeks after transplantation,the mean NAA/Cr,Glu/Cr and mI/Cr in three groups were 1.13 ±0.07,0.86 ±0.05 and 1.14 ±0.05,0.45 ± 0.04,0.38 ± 0.02 and 0.44 ± 0.03,0.58 ± 0.04,0.67 ± 0.04 and 0.53 ± 0.04,respectively,and differences were statistically significant ( F =112.092,23.076,44.367,P ＜ 0.01 ).NAA/Cr and Glu/Cr ratios were increased and mI/Cr was decreased in NSCs group,and the difference was significant compared with PBS group at the same time point ( t =0.271,0.071,0.089,P ＜ 0.05 ).There were no significant differences in NAA/Cr and Glu/Cr ( t =0.013,0.012,P ＞ 0.05 ),but there was a significant difference in mI/Cr between NSCs and control groups ( t =0.046,P ＜ 0.05).There were no significant differences in Cho before and after transplantation among the three groups (P ＞ 0.05 ). Nissl's staining showed that the number of neurons in the hippocampal area increased more significantly in tg mice receiving NSCs than that without receiving NSCs.Electron microscopy showed that most hippocampal NSCs in NSCs group were morphologically normal with abundant organelles,while hippocampal NSCs in PBS group were swollen with sparse synapses.Conclusion 1H-MRS is able to display intracranial metabolite changes before and after NSCs in APP-PS1 double transgenic AD mice and has an applicable value in evaluating the therapeutic effect of NSCs on AD.

Objective To label neural stem cells (NSCs) with superparamagnetic iron oxides (SPIO) and to explore the tropism of NSCs after transplantation into the hippocampus of APP/PS1 AD mice by MRI.Methods NSCs from C57BL/6 mouse were cultured and identified.Feridex and Poly-L-Lysine were added into the medium to be co-cultured to make magnetic labeled NSCs and transmission electron microscopy was used to identify the iron particles in NSCs.Transgenic (tg) and wild-type (wt) mice at 12 months of age were divided into three groups: SPIOs labeled NSCs group (A and C),unlabeled NSCs group(B).Feridex-labeled NSCs were migrated into the hippocampus of APP/PS1 AD mice to monitor in vivo by MRI.After 1,2,4 and 6 weeks,the mice were sacrificed and their brain tissues were sectioned to investigate the migration of SPIO labeled NSCs and compared with MRI.Results NSCs of C57BL/6 mice were cultured successfully.Transmission electron microscope showed visible iron granules in cytoplasm.MRI detection of labeled cells: T2WI and T2* WI showed remarkable low signal intensity at the hippocampus injection points 1 week after transplantation,particularly on T2* WI.Area of low signal intensity enlarged increasingly along the injection points after 2 weeks.At 4 weeks,area of low signal intensity spread throughout the hippocampus,but intensity shadowed Six weeks later,low signal intensity almost disappeared.There was no obvious low signal change in unlabeled cell transplantation group.For wt mice,size and location of low signal did not appear obvious change at all designated time points.Prussian blue positive cells were observed in the hippocampus,indicating that NSCs labeled with SPIO could survive,migrate and differentiate in the brain of the APP/PS1 AD mice.Changes of pathology were well correlated with the area where a signal intensity loss was observed in MRI 1,2,4 and 6 weeks after transplantation Conclusions Diffuse migration of transplanted NSCs labeled with SPIO is observed in the hippocampus in APP/PS1 tg mice,and MRI technique is an ideal method for tracking labeled stem cells after grafting in vivo.

AIM To speculate the hypoglycemic mechanism for rats with type 2 diabetes by exploring the therapeutic effects of leaf aqueous extract from Cyclocarya paliurus on liver insulin receptor (InsR) and insulin receptor substrate 2 (IRS-2).METHODS The diabetic rat model was established through intraperitoneal injection of streptozotocin and fed with high-fat diet.The moleled rats were equally assigned into the control group and leaf aqueous extract from Cyclocarya paliurus group (extract group).After the test extract was orally administrated for four weeks,body weight,urine output,food intake,water intake and fasting blood-glucose (FBG) were measured,and the levels of serum insulin,InsR and IRS-2 mRNA in liver tissue were investigated in rats.RESULTS Compared with the control group,the extract group showed a reduction in urine output,food intake,water intake,FBG and insulin levels.Meanwhile,the rats' body weights in extract group were presented a trend to increase.The gene expressions of InsR and IRS-2 in liver tissue were up-regulated.Moreover,the insulin sensitivity was improved.CONCLUSION The leaf aqueous extract from Cyclocarya paliurus can reduce FBS,improve insulin sensitivity,which may be associated with the increase of InsR and IRS-2 gene expression in liver tissue.

Objective To develop a new method for correction of the deformity of the complete bilateral cleft lip with short pre-lip. Methods Two specimens with the complete bilateral cleft lip was chosen to investigate the arterial supply to the pre-lip. The arteries were observed by radiography when the specimen was infused with lead oxide. 16 patients with the complete bilateral cleft lip and shortage in the length of pre-lip were enrolled in this study. Taking advantage of the unique character of arterial supply of complete bilateral cleft lip, those with short pre-lip were corrected. Photos of them were taken when preand post-operation for further measurement. And more than 1 year following-up was conducted to judge if the results were good enough or not. 30 babies were chosen as a control group to confirm the results of surgury objectively. Results Post nasal septal arteries were found to supply the pre-lip of the bilateral complete cleft lip. Appearance of 16 patients was good after surgery or 1 years later. There was no statistical significant difference between the patients and control group in the measurement data but nasal-lip angle. Conclusion It is an effective way to correct deformity of the bilateral cleft lip with short pre-lip by taking advantage of post nasal septal arterie's flap.

Objective　 To study the effects of sodium butyrate on proliferatio n, differentiation and apoptosis of immortalized esophagus epithelial cells. Methods　 SHEE, an immortalized human fetal esophageal epithelial cell line induced by HPV18 E6E7, was cultivated in culture flasks and 24-well plates. Two experi m ent groups of cultured cells were treated with 1 and 5 mmol/L of sodium butyrate respectively for 4 days, and one group of untreated cells set aside as control. The numbers of cloned cells were calculated. The ultra-structure of SHEE cells was examined by transmission electron microscopy (TEM). The cell cycle and numbe r of apoptotic cells were measured by flow cytometry, Ki-67 and cytokeratin of ce lls were detected by immunohistochemistry method and F-actin of cells labeled w ith phalloidin was examined by laser confocal scanning microscopy. Results　 Colo ny formations showed a significant decrease in the 2 experiment groups after 4 d ays of culture (P<0.01). In the 1 mmol/L group, the cells at S phase were di minish ed and arrested at G0/G1 phase. Compared with control group, Ki-67 positive cells were found decreased, while F-actin and cyto keratin were increased. Apoptotic cells in 5 mmol/L group were increased markedl y. Conclusions　 Sodium butyrate may induce SHEE cells growth arrest,differentiation and apoptosis. The effects depend on sodium butyrate concentrat ion and time of exposure. Whether it can be used in combination with other antic ancer drugs should be further studied.

OBJECTIVEStudy on the promoter effects of sodium butyrate in high or low dosages on carcinogenesis process, based on the immortalization of human fetal esophageal epithelium induced by human papillomavirus (HPV) 18E(6)E(7) genes.

METHODSThe immortalized esophageal epithelium SHEE was treated with high concentration of the sodium butyrate (80 mmol/L) and then with low concentration (5 mmol/L) for 8 weeks respectively. The cells were cultured continuously without sodium butyrate for 14 weeks. The morphology, proliferation and apoptosis of the cells were studied by phase contrast microscopy, immunohistochemistry and flow cytometry. The dead and the viable cells were assayed by fluorescent microscopy with Hoechst 33342 and Propidium iodide staining. Tumorigenesis of the cells was assessed by soft agar colony formation and by transplantation of cells into nude mice and SCID mice.

RESULTSWhen cells were exposed to high concentration of sodium butyrate, cell death was increased leaving few live cells. When cells were cultured in the medium with low concentration of sodium butyrate, the first proliferative stage appeared. Removal of the butyrate caused the cell to enter a crisis stage with a long doubling time resembling senescent cells. After the crisis stage, the cells progressed to the second proliferation stage with continuous replication and atypical hyperplasia. At the end of the second proliferative stage, carcinogenesis of the cells appeared with large colonies in soft-agar and tumor formation in transplanted SCID mice and nude mice.

CONCLUSIONSThe malignant change of the immortalized epithelium by the effects of sodium butyrate is the consequence of a two-stage mortality mechanism: cells death by butyrate cytotoxicity and cell crisis by abrogation of sodium butyrate. These data reveal that in high dosage, sodium butyrate induces cell death and in low dosage, it induces cell proliferation, which emphasizes the importance of butyrate as a promotor of carcinogenesis.

Animals ; Butyrates ; toxicity ; Carcinogens ; toxicity ; Cell Death ; drug effects ; Cell Division ; drug effects ; Cell Line, Transformed ; Cell Transformation, Neoplastic ; chemically induced ; Esophageal Neoplasms ; etiology ; Esophagus ; pathology ; Humans ; Mice ; Mice, Inbred BALB C ; Papillomaviridae ; pathogenicity

Objective:To discuss the pathological relationship between P33ING1 and stomach cancer and its clinical significance. Methods: In 71 cases of stomach cancer specimen, twelve cases of gas tric mu cous membrane atypical hyperplasia tissues and 18 cases of normal gastric mucous membrane tissue（as control）,the expression of P33ING1 were detected b y EnVision immunohistochemical method,while the expression of P53 and Bcl -2 in stomach cancer were also detected. Results: P33IN G1 expression in mucous membrane atypical hyperplasia group and control group was positive, the expression in stomach cancer group was extremely low(62.0%，44 /71), significantly different from the other 2 groups(P<0.01).P33ING1 expression in stomach cancer was related to the tumor growth, lymph node meta s tasis and tumor polarization (P<0.01), P53 expression was related to tumor s ize, growth and lymph node metastasis (P<0.01). Bcl-2 expression was relate d to lymph node metatasis and tumor polarization.The expression of P33ING1 was related to that of P53 in stomach cancer(P<0.05),while had no relation with that of Bcl-2.Conclusion:P33ING1 may play an importa nt role in the occurrance and development of stomach cancer.It's very important to detect the expression of P33ING1 and P53 simultaneously.