To observe the expression of N - cadherin and fibronectin in retinal pigment epithelium ( RPE) cells in vitro under high glucose conditions, furthermore, to explore the effects of high glucose on epithelial -mesenchymal transition (EMT) in RPE cells.
●METHODS: Human RPE (hRPE) cells were cultured in vitro. Containing a final concentration of 60mmol/ L glucose was used for high glucose treatment. The cells were divided into normal glucose group (5. 5mmol/ L, NG) and high glucose group (24, 48 and 72h) respectively. The expression of N - cadherin and fibronectin in hRPE cells were evaluated by immunofluorescence and real -time PCR.
●RESULTS:RPE cells became disorganized and swollen over time under high glucose conditions, especially in 72h subgroup. lmmunohistochemical analysis revealed that the expression of N - cadherin in RPE cells under high glucose conditions was decreased compared with that in the control group, while the expression of fibronectin was increased. Real - time PCR results showed that the expression of N - cadherin mRNA in high glucose group was decreased at 24h compared with that in the control group, and declined markedly at 72h ( F = 12. 252, P =0. 000). There were no significant differences between the control group and the high glucose group at 24h, while the differences between the control group and the high glucose group (48 and 72h) were significant respectively (P < 0. 05 ). Meanwhile, the expression of fibronectin mRNA in RPE cells was increased in high glucose group at 24h, and reached the peak at 72h (F = 50. 543, P = 0. 000). There were no significant differences between the control group and the high glucose group at 24h. Compared with the control group, the expression of fibronectin mRNA in hRPE cells was increased significantly in high glucose group (48 and 72h) respectively (P= 0. 000, P= 0. 000).
●CONCLUSlON: The expression of epithelium marker N-cadherin is down - regulated under high glucose conditions in hRPE cells in vitro. Meanwhile, the expression of mesenchymal maker fibronectin is induced and appeared to EMT changes. Results of this study will enrich our growing understanding in proliferative diabetic retinopathy and hopefully lead to novel insights for the pathogenesis and therapeutic treatments.

Objective To evaluate the effect of transurethral feedback microwave thermotherapy with the ProstaLund CoreTherm Device(PLFT) in high risk patients with benign prostate hyperplasia (BPH). Methods Sixty-six high risk patients diagnosed with BPH, including aged ≥80 in 32 pa-tients, hypertension in 31 patients, diabetes in 5 patients, heart failure in 8 patients, chronic obstruc-tive pulmonary disease in 8 patients, cerebral infarction in 11 patients, fracture, amputation or joint stiffness unsuitable for lithotomy position in 3 patients, abnormal blood coagulation in 4 patients, pan-creatitis in 2 patients, cardiac arrhythmia in 6 patients and malignant tumor in 3 patients, were treated with PLFT using individual power at urethral local anesthesia, resulting in coagulation necrosis in 15%-30% of prostate tissue around urethra. Meanwhile, real-time monitoring the temperature of prostate and the tissue around it was used. All patients were evaluated by comparing volume of pros-tate, maximal urinary flow (Q_max), international prostate symptom score (IPSS) and quality of life questionnaire (QOL) in pre-treatment and three months after respectively. Results All of patients well tolerated PLFT. There was bleeding lightly, infection lightly and temporary incontinence. There was no severe surgical complication. After three months, the volume of prostate reduced from 62. 2 ml to 44.5 ml; IPSS decreased from 23. 4 to 11.7; QOL decreased from 4.5 to 2.4; Q_max rised from 4, 2 ml/s to 11.2 ml/s. All differences reached significance. Conclusion PLFT is one of effective and safe treatments for patients with BPH especial BPH complicating with severe conditions.

Objective To investigate the approach and safety of minimally invasive surgical procedures treating symptomatic caliceal diverticular calculi. Methods Clinical data of 21 cases with symptomatic caliceal diverticular calculi were retrospectively reviewed. Twelve females and 9 males aged 22 to 57 years old. The average diameter of caliceal diverticulum was 3.7 cm (2.5-7.0 cm) and average diameter of calculi was 2.3 cm (0.8-3.5 cm). The patients underwent flexible ureteroscopic lithotripsy, PCNL or mPCNL, laparoscopic techniques and laparoscopy-assisted transperitoneal PCNL, respectively. Four cases underwent flexible ureteroscopic lithotripsy. PCNL (2 cases) or mPCNL(5 cases)were performed in 7 cases. Nine cases underwent laparoscopic techniques. Laparoscopyassisted transperitoneal PCNL was performed in 1 case. Results The operations were performed successfully in 21 cases. No case need to transfer to open surgery during the operation and no major complications like perforation or organic injury were noted. One case with iatrogenic arteriovenous fistula of the kidney after 1 week postoperative was cured by delayed hemorrhage 2 days later, while clinical symptoms of 2 cases with residue calculi relieved. 19 cases without residue calculi were followed up for 6 to 12 month without recurrence. Conclusions After handling indication of treatment efficiently and creating advisable therapeutic decision-making, minimally invasive surgical procedures treating symptomatic caliceal diverticular calculi appears to be effective and safe.

To study the equipment modular allocation in the emergency medical unit after Beijing emergency public affairs. Using an improved Delphi method, 25 experts from various fields in Beijing were involved into two rounds consultations. The questionnaire of the first round consultation was formed based on analysis of emergency rescue equipment. Based on the result of the first round consultation, the questionnaire of the second round consultation was set up. The two rounds of questionnaires had the responding rates of experts 100% respectively, and the au-thoritative coefficients of the experts were 0.81 and 0.88. An emergency medical rescue equipment solution with three de-grees modular configuration in Beijing was made based on the consultations and experts' advices. The solu-tion develops the emergency medical service ability of mobile medical units and vehicle-mounted mobile hospital. LUO Ying and GAO Hong-wei are the first authors who contributecl equally to the article.

OBJECTIVETo identify the predisposing factors of deep neck abscess and review diagnosis and treatment experience.

METHODSA respective review was conducted in 50 cases who were diagnosed as having deep neck abscess in this hospital from Jan. 1997 to Dec. 2002.

RESULTSThe causes of deep neck abscess were tooth diseases (3 cases), acute tonsillitis and laryngitis (8 cases), infection of upper respiratory tract (9 cases), foreign bodies in esophagus (14 cases), diabetes mellitus (5 cases), uncertain cause (11 cases). Among 21 cases of pus bacterial cultivation, 13 (64%) cases were positive. By different ways of drainage including neck-mediastinum incision, and use of large dosage of antibiotics, 46 six cases were cured and 4 cases died. Two died of massive hemorrhage of neck blood vessel burst, one massive hemorrhage of upper digestive tract, another infective shock.

CONCLUSIONSOnce deep neck abscess is diagnosed, early surgical drainage, appropriate use of antibiotics and control of complications and accompanying diseases are very important to improve the cure rate.

Adolescent ; Adult ; Aged ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Male ; Middle Aged ; Neck ; Retropharyngeal Abscess ; Retrospective Studies ; Soft Tissue Infections ; Young Adult

Objective To evaluate the effect of transurethral feedback microwave thermotherapy with the ProstaLund CoreTherm Device (PLFT) in benign prostate hyperplasia ( BPH ) patients with high risk factors 24 months after treatment.MethodsSixty-two BPH cases with high risk factors of aged ≥ 80or complicating severe conditions of no less than one organ or system,were treated with PLFT under urethral local anesthesia.The average pre-treatment prostate volume,international prostate symptom score (IPSS),quality of life score (QOL) and maximal urinary flow (Qmax) were 62.03 ml,23.19,4.58 and 4.33 ml/s,respectively.The changes of prostate volume,IPSS,QOL and Qmax at 3 months,12 months and 24 months after treatment were analyzed.ResultsAll patients tolerated well of PLFT performed in common therapy room except lightly bleeding,minor infection and temporary incontinence.There was no severe surgical adverse event.After 3 months,the prostate volume reduced to 43.85 ml,IPSS decreased to 11.63,QOL decreased to 2.44,Qmax rose up to 11.44 ml/s； The average values were 45.10 ml,12.23,2.61 and 10.91ml/s at 12 months after treatment.The corresponding values were 45.80 ml,12.37,2.66 and 10.82 ml/s,respectively at 24 months after treatment.Compared with pre-treatment,all the parameters showed significant improvement ( P ＜ 0.01 ).ConclusionsPLFT is one of the effective and safe treatment options for BPH patients with high risk factors.It can be safely used on day-surgery patients.The best effect appears at 3 months after treatment.

The LEAFY (LFY) homologous gene of Dendrobium moniliforme (L.) Sw. was cloned by new primers which were designed based on the conservative region of known sequences of orchid LEAFY gene. Partial LFY homologous gene was cloned by common PCR, then we got the complete LFY homologous gene Den LFY by Tail-PCR. The complete sequence of DenLFY gene was 3 575 bp which contained three exons and two introns. Using BLAST method, comparison analysis among the exon of LFY homologous gene indicted that the DenLFY gene had high identity with orchids LFY homologous, including the related fragment of PhalLFY (84%) in Phalaenopsis hybrid cultivar, LFY homologous gene in Oncidium (90%) and in other orchid (over 80%). Using MP analysis, Dendrobium is found to be the sister to Oncidium and Phalaenopsis. Homologous analysis demonstrated that the C-terminal amino acids were highly conserved. When the exons and introns were separately considered, exons and the sequence of amino acid were good markers for the function research of DenLFY gene. The second intron can be used in authentication research of Dendrobium based on the length polymorphism between Dendrobium moniliforme and Dendrobium officinale.
Amino Acid Sequence ; Base Sequence ; DNA, Plant ; genetics ; Dendrobium ; genetics ; Exons ; Introns ; Orchidaceae ; genetics ; Phylogeny ; Plant Leaves ; genetics ; Plant Proteins ; genetics ; Plants, Medicinal ; genetics ; Sequence Alignment ; Sequence Homology, Amino Acid

OBJECTIVEThis study was to explore the cytotoxic effect and the related injury mechanism of deoxynivalenol (DON) on articular chondrocytes in human embryo.

METHODSArticular cartilage cells were isolated from knees of human embryo and cultured in DMEM/F12 medium. The cells of the 4th generation were divided into five groups and incubated with varying concentrations of DON as the followings: control group and group with DON of 0.1, 0.2, 0.4, 1.0 µg/ml. The effects of DON were observed 72 hours after incubation. Cell apoptosis was assayed by flow cytometry (FCM) with Annexin V-FITC/PI staining; MMP-13 and PGE2 were detected by ELISA kits; NO was measured by Griess assay with spectrophotometer. Inducible nitric oxide synthase (iNOS) and collagen II in cells were detected by FCM. The expression levels of iNOS, mRNA and collagen II mRNA were measured with RT-PCR.

RESULTSThe rates of cell apoptosis in DON groups were 6.78% - 19.05%, which were significantly higher than that in control (1.20%, F = 174.761, P < 0.05). The levels of NO in DON groups were 20.8 - 40.7 µmol/L, which were significantly higher than that in control (10.2 µmol/L, F = 91.966, P < 0.05). The levels of MMP-13 in DON groups were 0.25 - 0.56 µmol/L, which were significantly higher than that in control (0 µmol/L, F = 78.420, P < 0.05). The levels of PGE2 in DON groups were 3.2-20.6 µmol/L, which were significantly higher than that in control (11.6 µmol/L, F = 276.453, P < 0.05). The proportions of cells with positive iNOS in DON groups were 14.8% - 56.8% which were significantly higher than that in controls (7.1%, F = 214.614, P < 0.05). The proportions of cells with positive collagen II in groups with DON of 0.4 µg/ml and 1.0 µg/ml were 56.7% and 52.7%, which were significantly lower than that in control (62.2%, F = 5.134, P < 0.05). The relative absorbance values of iNOS mRNA in DON groups were 1.07 - 1.33, which were significantly higher than that in control (0.62, F = 8.358, P < 0.05). The levels of collagen II mRNA in groups with DON of 0.4 µg/ml and 1.0 µg/ml were 0.83 and 0.82, which were significantly lower than that in control (1.14, F = 7.887, P < 0.05).

CONCLUSIONDON could promote anabolism of NO in articular cartilage cells by which up-regulated the expression of PGE2 and MMP-13, which both promoted resolution of articular cartilage matrix such as collagen II. DON induced apoptosis in articular cartilage cells.

Cartilage, Articular ; cytology ; embryology ; Cells, Cultured ; Chondrocytes ; drug effects ; metabolism ; Dinoprostone ; metabolism ; Humans ; Matrix Metalloproteinase 13 ; metabolism ; Nitric Oxide ; biosynthesis ; Trichothecenes ; toxicity

Bletilla striata has been used as traditional Chinese medicine for several centuries. In recent years, the quality and quantity of wild B. striata plants have declined sharply due to habitat deterioration and human over-exploitation. Therefore, it is of great urgency to evaluate and protect B. striata wild plant resource. In this study, sequence-related amplified polymorphism (SRAP) markers were applied to assess the level and pattern of genetic diversity in twelve populations of B. striata. The results showed a high level of genetic diversity (PPB = 90.48%, H = 0.349 4, I = 0.509 6) and moderate genetic differentiation among populations (G(st) = 0.260 9). Based on the unweighted pair-group method with arithmetic average (UPGMA), twelve populations gathered in three clusters. The cluster 1 included four populations. There are Nanjing, Zhenjiang, Xuancheng and Hangzhou. The seven populations which come from Hubei Province, Hunan Province, Jiangxi Province and Guizhou Province belonged to the cluster 2. The cluster 3 only contained Wenshan population. Moreover, Mantel test revealed significant positive correlation between genetic distances and geographic distances (r = 0.632 9; P < 0.000 1). According to the results, we proposed a series of conservation consideration for B. striata.
China ; Genetic Markers ; Genetic Variation ; Genetics, Population ; Orchidaceae ; genetics ; Phylogeny ; Plants, Medicinal ; genetics

To identify adulterants from medicinal plants of Bletilla H. G. Reichenbach, the suitable candidate DNA barcoding of Bletilla was evaluated. In this study, the internal transcribed spacer (ITS) of nuclear ribosomal DNA, the LFY homologous gene intron 2 and chloroplast ycfl gene were amplified and sequenced from forty-one samples. The intra-specific and inter-specific divergences of Bletilla were calculated, and the identification efficiency was assessed using Barcoding Gap, NJ tree by K2P distance and BLAST1 method. The result showed the intra-specific divergence of nrDNA ITS and ycJfl (0.022-0.106 and 0.017-0.106) were obviously higher than the inter-specific divergence (0-0.012 and 0-0.015), and four species of Bletilla were also accurately distinguished in NJ trees. Whereas, there was no Barcoding Gap on LFY homologous gene intron 2, thus it cannot effectively identify species of Bletilla. Using NJ tree of nrDNA ITS and ycfl gene, powdery medicine and the adulterants of Bletilla were successfully unidentified. In conclusion, nrDNA ITS and ycfl can be used as a potential DNA barcoding to identify the medicinal plants in Bletilla and its adulterants. There were only three basic differences on nrDNA ITS between "Jujing baiji" and Bletilla striata of Lu'an in Anhui province, and two basic differences in ycfl. Based on morphological and molecular data, "Jujing baiji" could be recognized as the species of Bletilla striata.
Base Sequence ; DNA Barcoding, Taxonomic ; DNA, Plant ; genetics ; DNA, Ribosomal Spacer ; genetics ; Orchidaceae ; classification ; Plants, Medicinal ; classification