Objective To detect the mutations of ATP2C1 gene in patients with Hailey-Hailey dis- ease (HHD).Methods PCR and direct sequencing were performed in 17 patients and 120 healthy controls to screen the mutations in the exons of ATP2C1 gene.Results Eight mutations were identified in nine probands, including three deletion mutations (nt1464-1487 del/nt1462-1485del,1523delAT,2375delTTGT),three splice site mutations (360—2A→G,1415—2A→T,2243+2T→C) and two missence mutations (C920T and G1942T).None of the above mutations was found in the controls.Conclusion Eight specific novel mutations were identified in nine probands of HHD,which could be causative factors of the disease.

On account of the dense cuticles of the fresh stem and the light, hard and pliable texture of the dried stem, Dendrobii Caulis is difficult to dry or pulverize. So, it is very important to the ancient doctors that Dendrobii Caulis should be properly treated and applied to keep or evoke its medicinal effects. The current textual research results about the preliminary processing, processing and usage methods of Dendrobii Caulis showed that: (1) In history the clinical use of fresh or processed Dendrobii Caulis as teas and tinctures were very common. (2) Its roots and rhizomes would be removed before using. (3) Some ancillary approaches were applied to shorten drying times, such as rinsing with boiling mulberry-ash soup, washing or soaking with liquor, mixing with rice pulp and then basking, etc. (4) According to the ancients knowledge, the sufficient pulverization, by means of slicing, rasping, hitting or pestling techniques, was necessary for Dendrobii Caulis to take its effects. (5) The heat processing methods for Dendrobii Caulis included stir-baking, stir-frying, steaming, decocting and stewing techniques, usually with liquor as an auxiliary material. Among above mentioned, steaming by pretreating with liquor was most commonly used, and this scheme was colorfully drawn in Bu Yi Lei Gong Pao Zhi Bian Lan (Ming Dynasty, 1591 CE) ; moreover, decocting in advance or long-time simmering so as to prepare paste products were recommended in the Qing Dynasty. (6) Some different processing programs involving stir-baking with grit, air-tightly baking with ondol (Kangs), fumigating with sulfur, which appeared in modern times and brought attractive outward appearance of the drug, went against ancients original intentions of ensuring drug efficacy.
Dendrobium ; History, Ancient ; Medicine, Chinese Traditional ; history ; Technology, Pharmaceutical ; history

Adolescent ; Adult ; Aged ; Biopsy ; Delayed Graft Function ; diagnosis ; pathology ; Female ; Graft Rejection ; diagnosis ; pathology ; Humans ; Kidney ; pathology ; physiopathology ; Kidney Function Tests ; Kidney Transplantation ; pathology ; Male ; Middle Aged ; Young Adult

BACKGROUNDCerebral ischemia-reperfusion injury is the main reason for the loss of neurons in the ischemic cerebrovascular disease. Therefore, to deeply understand its pathogenesis and find a new target is the key issue to be solved. This research aimed to investigate the neuroprotective effects of salvianolic acid B (SalB) against oxygen-glucose deprivation/reperfusion (OGD/RP) damage in primary rat cortical neurons.

METHODSThe primary cultures of neonatal Wister rats were randomly divided into the control group, the OGD/RP group and the SalB-treatment group (10 mg/L). The cell model was established by depriving of oxygen and glucose for 3 hours and reperfusion for 3 hours and 24 hours, respectively. The neuron viability was determined by MTT assay. The level of cellular reactive oxygen species (ROS) was detected by fluorescent labeling method and spin trapping technique respectively. The activities of neuronal Mn-superoxide dismutase (Mn-SOD), catalase (CAT) and glutathione peroxidase (GSH-PX) were assayed by chromatometry. The mitochondria membrane potential (ΔΨ(m)) was quantitatively analyzed by flow cytometry. The release rate of cytochrome c was detected by Western blotting. The neuronal ultrastructure was observed by transmission electron microscopy. Statistical significance was evaluated by analysis of variance (ANOVA) followed by Student-Newman-Keuls test.

RESULTSOGD/RP increased the level of cellular ROS, but decreased the cell viability and the activities of Mn-SOD, CAT and GSH-PX; SalB treatment significantly reduced the level of ROS (P < 0.05); and enhanced the cell viability (P < 0.05) and the activities of these antioxidases (P < 0.05). Additionally, OGD/RP induced the fluorescence value of ΔΨ(m) to diminish and the release rate of cytochrome c to rise notably; SalB markedly elevated the level of ΔΨ(m) (P < 0.01) and depressed the release rate of cytochrome c (P < 0.05); it also ameliorated the neuronal morphological injury.

CONCLUSIONThe neuroprotection of SalB may be attributed to the elimination of ROS and the inhibition of apoptosis.

Animals ; Apoptosis ; drug effects ; Benzofurans ; pharmacology ; Catalase ; metabolism ; Cells, Cultured ; Cerebral Cortex ; blood supply ; Cytochromes c ; secretion ; Glutathione Peroxidase ; metabolism ; Hypoxia-Ischemia, Brain ; drug therapy ; Membrane Potential, Mitochondrial ; drug effects ; Neuroprotective Agents ; pharmacology ; Rats ; Rats, Wistar ; Reactive Oxygen Species ; metabolism ; Reperfusion Injury ; prevention & control ; Superoxide Dismutase ; metabolism

OBJECTIVETo explore the effect of three-dimensional structure of dermal matrix on biological behavior of fibroblasts (Fb) in the microcosmic perspective.

METHODSThe three-dimensional structure of dermal tissue was analyzed by plane geometric and trigonometric function. Microdots structure array with cell adhesion effect was designed by computer-assisted design software according to the adhesive and non-adhesive components of dermal tissue. Four sizes (8 microm x 3 microm, space 6 microm; 16 microm x 3 microm, space 6 microm; 16 microm x 5 microm, space 8 microm; 20 microm x 3 microm, space 2 microm) of micropier grid used for cell culture (MPGCC) with cell-adhesive microdots, built up with micro-pattern printing and molecule self-assembly method were used to culture dermal Fb. Fb cultured with cell culture matrix without micropier grid was set up as control. The expression of skeleton protein (alpha-SMA) of Fb, cell viability and cell secretion were detected with immunohistochemistry, fluorescent immunohistochemistry, MTT test and the hydroxyproline content assay.

RESULTSThe three-dimensional structure of dermal tissue could be simulated by MPGCC as shown in arithmetic analysis. Compared with those of control group [(12 +/- 3)% and (0.53 +/- 0.03) microg/mg, (0.35 +/- 0.04)], the expression of alpha-SMA [(49 +/- 3)%, (61 +/- 3)%, (47 +/- 4)%, (51 +/- 3)%] and the content of hydroxyproline [(0.95 +/- 0.04), (0.87 +/- 0.03), (0.81 +/- 0.03), (0.77 +/- 0.03) microg/mg] were increased significantly (P < 0.05), the cell viability of Fb (0.12 +/- 0.03, 0.13 +/- 0.04, 0.14 +/- 0.03, 0.19 +/- 0.03) cultured in MPGCC was decreased significantly (P < 0.05). When the parameters of micropier grid were changed, the expression of alpha-SMA, the cell viability and the content of hydroxyproline of Fb cultured in four sizes of MPGCC were also significantly changed as compared with one another (P < 0.05).

CONCLUSIONSMPGCC may be the basic functional unit of dermal template, or unit of dermal template to call. Different three-dimensional circumstances for dermal tissue can result in different template effect and wound healing condition.

Biocompatible Materials ; Cell Adhesion ; Cell Culture Techniques ; Cells, Cultured ; Computer-Aided Design ; Dermis ; Fibroblasts ; cytology ; Tissue Engineering ; methods

OBJECTIVETo research the function of endophytes of mistletoe in parasitism process of mistletoe in Pterocarya stenoptera.

METHODEndophytes from eight different parts of the mistletoe were separated by explant culture, and further screened by different CMC plates culture and DNS method to get cellulase high productive strains. The distribution of the endophytic fungus parasitized in mistletoe were prepared and stained to demonstrate by histological section of the intumescentia part of the P. stenoptera.

RESULTThe histological section indicated that aboundent of hyphasma were distributed around the haustorium of the mistletoe. Eighty three strains of endophytic fungus were separated, 38 of them were able to degrade cellulose, 19 strains showed high cellulase activity and 10 of which were separated from the parasitic position.

CONCLUSIONEndophytic fungus of mistletoe can secrete cellulase and assist the haustorium of mistletoe to breakthrough the cell walls as well as intercellular space tissues of the P. stenoptera, thus, the endophytic fungus plays an important role in the parasitism process of mistletoe in P. stenoptera.

Cellulase ; metabolism ; Cellulose ; metabolism ; Fungi ; metabolism ; Juglandaceae ; Symbiosis ; Viscum ; cytology ; microbiology

OBJECTIVETo explore the effect of 18-β glycyrrhetinic acid (GA) on the endoplasmic reticulum of nasal epithelial cells in allergic rhinitis (AR) model rats.

METHODSTotally 96 Wistar rats were randomly divided into the blank group, the AR model group, the loratadine group, the GA group, 24 in each group. AR models were established by peritoneally injecting ovalbumin (OVA). Morphological scoring was performed. GA at 21. 6 mg/kg was intragastrically administered to rats in the GA group. Nasal mucosal tissues were taken for electron microscopic examinations at the second, fourth, sixth, and tenth week after drug intervention.

RESULTSThe overlapping score was 2.10 ± 0.45 in the blank group, 5.10 ± 0.56 in the loratadine group, 5.10 ± 0.56 in the AR model group, 5.20 ± 0.78 in the GA group, showing statistical difference when compared with the blank group (P < 0.01). Results under transmission electron microscope showed that the number of the endoplasmic reticulum increased in the AR model group, with obvious cystic dilatation, a lot of vacuole formation, and degranulation. A large number of free ribosomes could be seen in cytoplasm. With persistent allergen exposure, changes mentioned above was progressively aggravated in the endoplasmic reticulum of nasal mucosal epithelium in the AR model group. But the dilation of endoplasmic reticulum, vacuole formation, and degranulation were relieved in the GA group, and got close to those of the blank group.

CONCLUSION18-β GA could improve the expansion, vacuolization, and degranulation of the endoplasmic reticulum of nasal epithelial cells in AR model rats.

Animals ; Anti-Inflammatory Agents ; pharmacology ; therapeutic use ; Endoplasmic Reticulum ; Epithelial Cells ; drug effects ; Glycyrrhetinic Acid ; pharmacology ; therapeutic use ; Nasal Mucosa ; drug effects ; Rats ; Rats, Wistar ; Rhinitis, Allergic ; drug therapy

Objective To estimate the intangible cost and associated factors on patients with hepatitis B-related diseases, so as to explore the differences of the three elicitation techniques on the health economics-related information by trained investigators, using a structured questionnaire. WTP was employed to estimate the intangible cost while an open-ended question format, together with iterative bidding game and payment card were respectively used to elicit WTP for the hypothetical cure of hepatitis B-related diseases. A Multiple linear stepwise regression model was determined to identify those factors potentially affecting the intangible cost. Results A total of 564 subjects from 641 patients with hepatitis B-related diseases were identified for the inclusion of this study. The average annual intangible cost of patient with hepatitis B-related diseases was 54 320.4 Yuan (Ren Minbi).The intangible cost accounted for 53.0% of the total cost, which was much more than the proportions of the direct and indirect costs (38.5% and 8.5%, respectively). Among annual personal and the household income of the patient, proportions of intangible cost were 262.6% and 67.6% respectively,suggesting that the patients were under huge spiritual and psychological pressure. Response rate of the approach, combined open-ended questions with iterative bidding game, was the highest (76.6%) among the three elicitation formats. Considered the characteristics of data being gathered, the approach seemed to be more reasonable. Further studies were needed to examine the results yielded from other WTP elicitation formats. We also noticed that the progression of disease was associated with the increase of direct and indirect costs, but not with the intangible cost. Data from the multiple linear stepwise regression analysis indicated that the types of hospital and commercial medical insurance were significantly different in explaining the variation of the intangible cost. Conclusion Measures should be taken to reduce the intangible cost of hepatitis B-related diseases. The approach regarding the combination of open-ended questions with iterative bidding game should be recommended when carrying our further WTP studies of this kind.

Objective To summarize the experience of long-term survival in patients after simulta- neous kidney-pancreas transplantation(SKPT)with modified enteric drainage(ED).Methods From October 2001 to July 2004,6 patients with end-stage renal disease due to Type 1 diabetes underwent SKPT with modified ED,ie,side-to-side anastomosis between the duodenum of donors and jejunum of recipients. The medication regimen included:mycophenolic acid 500 mg and tacrolimus 2 mg before operation;methyl- prednisolone(MP)1.0 during operation;and 2-dose anti-IL-2 receptor monoclonal antibody(2 cases)or antihuman thymocyte globulin(ATG)(4 cases)for immune induction therapy;MP was used on the first 3 d after transplantation,triple immunosuppressive therapy(tacrotimus,mycophenolic acid and prednisone)was used on the second d after transplantation.Anticoagulants such as low molecular heparin or alprostadil were used for 7-10 d to prevent thrombosis in pancreas graft.Somatostatin was used as prophylaxis for graft pan- creatitis.Ganciclovir was used to prevent cytomegalovirus infection when renal graft gradually recovered 3 to 5 d after transplantation.The follow-up was from 1 year and 3 months to 4 years and 1 month.Results Transplantation was successful in all 6 cases.The blood sugar levels were 6-16 mmol/L.Low-dose insulin was used for 5-10 d,then the blood sugar levels returned to normal range.One of 6 patients experienced nephrotoxicity because of high tacrolimus blood concentration at 7 d after operation;after 3 dialyses and re- duction of tacrolimus dose,the renal allograft regained normal function.Three cases experienced alimentary tract hemorrhage at 14,20 and 22 d,respectively,after operation;the bleeding was stopped after treatment. There were no complications such as pancreatic fistula,intestinal fistula and thrombosis early after operation. All the patients are now alive,specifically,1 survived over 4 years,3 over 3 years,1 over 2 years,and 1 over 1 year.All had normal blood sugar free of insulin use.Five cases had normal renal graft function,with normal sCr,and 1 had sCr＞400?mol/L. Two cases were admitted to hospital due to upper respiratory infection and furuncles in the skin of head 6 months and 2 years,respectively,after operation.They were both cured.No complications such as urinary infection,metabolic acidosis and dehydration occurred.Conclusions SKPT is effective for the treatment of end-stage renal disease due to Type 1 diabetes.SKPT with modified ED are relatively simple with physiological compatibility and fewer complications.High quality of donated organs, HLA matching,pancreatic drainage pattern,rational periopcrative medications and infection late after trans- plantation are important factors affecting the long-term survival of the patients.

Adolescent ; Adult ; China ; epidemiology ; Disease Outbreaks ; Female ; Humans ; Incidence ; Leptospirosis ; epidemiology ; mortality ; Male ; Middle Aged