Objective To identify a rapid and efficient fungal genomic DNA extraction method for PCR amplification.Methods Genomic DNA was extracted from Penicillum marneffei,Rhizopus microsporus,Cryptococcus neoformans and Candida albicans by heating pyrolysis,microwave,repeated freezing and thawing,lysozyme digestion,overnight snail enzymatic and Qiagen kit methods.DNA electrophoretogram was observed by gel imaging system.The concentration and purity of extracted DNA were determined with an ultramicro nucleic acid protein tester and the yields were calculated.PCR amplification and sequencing were also performed.ANOVA and SNK-q test were used for data analysis.Results There were statistical differences in concentrations and yields of the fungal DNA extracted from Penicillum marneffei (hyphal phase and yeast phase),Rhizopus microsporus,Coptococcus neoformans and Candida albicans by six methods (F=750.83,220.95,669.35,132.01,510.20 and 1658.35,287.10,963.64,1147.77,4521.22,all P ＜0.01).Of six methods,microwave method gained the highest DNA concentration and yield,followed by heating pyrolysis method,while Qiagen kit method obtained the lowest concentration and yield.All DNA extracted by 6 kinds of methods were positive in PCR amplification.Conclusion All of the six methods can be used for fungal DNA extraction which is sufficient for PCR amplification,but microwave and heating pyrolysis methods are more easy and simple to perform.

Deafness ; diagnosis ; genetics ; Genetic Counseling ; Humans

Objective To primarily investigate the clinical value of analgesia/nociception index (ANI) in evaluating the analgesic effect during lobectomy performed via video-assisted thoracoscope.Methods Forty ASA physical status Ⅰ or Ⅱ patients,aged 25-64 yr,weighing 45-80 kg,undergoing elective lobectomy performed via video-assisted thoracoscope,were enrolled in this study.After induction of anesthesia with propofol,sufentanil and cisatracurium,patients received double lumen endotracheal intubation.Anesthesia was maintained with targetcontrolled infusion of propofol,and iv infusion of remifentanil and cisatracurium.The concentration of propofol was adjusted to maintain the bispectral index (BIS) value in the range of 40-60.ANI,HR,systolic blood pressure (SBP),diastolic blood pressure (DBP) and BIS value were recorded within 5 min before and after the predefined time points including posture change between lateral and supine position,ventilatory pattern change between onelung and double-lung ventilation,skin incision and trocars insertion,lymph node dissection and pleural lavage.At skin incision and during trocars insertion,lymph node dissection and pleural lavage,the development of hemodynamic responses (increase in HR and SBP ＞ 20％ of baseline value) were recorded.Results The incidence of hemodynamic responses was 100％ at skin incision and trocars insertion,and 84 ％ during No.4,7,10 groups of lymph node dissection and after pleural lavage and difference was found in ANI during these stimuli.ANI was significantly decreased within 5 min after skin incision,trocars insertion,No.4,7,10 groups of lymph node dissection and pleural lavage than that before the procedures (P ＜ 0.05).The BIS value was maintained at 40-60,and no significant changes were found between before and after the procedures (P ＞ 0.05).No significant changes were found in ANI,HR,SBP,and DBP between before and after the changes of posture and respiratory pattern (P ＞ 0.05).Conclusion ANI can be used to evaluate the analgesic effect during lobectomy performed via video-assisted thoracoscope in patients and is unaffected by the changes of posture and ventilatory pattern.

Aim To investigate the effects of Aplysin on the inhibition of gastric cancer cell in vitro .Methods MTT assay was used to examine the inhibition of gastric cancer cell 1ine SGC-7901 by Aplysin in different concentrations and at different times.The morphologic changes and the apoptosis of SGC-7901 was observed by inverted microscope and Hematoxylin-Eosin(HE)staining.Reverse transcriptase polymerase chain reaction(RT-PCR)assay was used to detect the changes of COX-2 mRNA expressions.Results Aplysin could decrease the proliferation significantly in a dose-dependent manner in SGC-7901 cells.When treating SGC-7901 with Aplysin in concentration of 120, 240 mg·L~(-1) for 24 h, the growth of the cell was obviously inhibited observing by inverted microscope.Aiso, when treating with the same concentration for 18 h, its chromatin became crimpled and breakdown, as well as cell shrinkage and apoptotic bodies formation when using HE staining.The apoptotic rates(%)of SGC-7901 was(15.0±2.12)%, (18.4±2.3)%, respectively, which was significantly higher than(1.4±0.55)% that in control group(P <0.01).60、120、240 mg·L~(-1) Aplysin could not effectively inhibited the mRNA expressions of COX-2(P ＞0.05).Conclusions Aplysin can inhibit the proliferation and induces apoptosis of SGC-7901 cells.

?Pathological myopia are often complicated by a series of pathological changes in fundus including foveoschisis, which can lead to visual dysfunction when processing with retinal detachment, macular hole, epiretinal membrane and vitreoretinal traction diseases. According to the current knowledge, the main mechanism of foveoschisisi might be attributed to the impaired macular structure and function caused by a variety of traction on the retinal and retina poor condition. Surgical treatments have been reported to be effective in treating foveoschisis, however, the indications and surgical procedures are still controversial. ln this article, we reviewed the clinical features, diagnosis, treatment strategies and prognosis of pathological myopia foveoschisis.

Objective To explore continuous renal replacement therapy(CRRT) machine for plasma exchange in critical disease in children.Methods Retrospective study of 8 patients(8 month to 14 years,mean 5.7 years) and 32 plasma exchange treatments,after(adowble) lumen catheter inserted into the subclayian venous,using the Baxter BM25 machine with commercially available plasma filters.Results Five patients(3 ABO-incompatibility in bone marrow transplantation,1 thrombotic thrombocytopaenic purpura TTP,1 sepsis) gained full recovery.One systemic lupus erythematosus(SLE) and 1 sepsis experienced moderate improvement while 1 case of acute disseminated encephalomyelitis failed PE treatment.The average total exchange volume was 80-100 mL/kg,achieved at a blood flow rate of 5-10 mL/(kg?min) and a turnover rate of 60-120 mL/(kg?h) over a 3-hours duration.Thirty-one PE treatments were finished smoothly,one of which experienced the serious complication involving plasma filter.Conclusion Plasma exchange therapy is a safe and effective procedure for severe autoimmune abnormalities and pathogen removal in children.

Clinical pathway (CP), as a standardized approach and clinical management process for disease diagnosing and treating, is being widely used with more relevant publications reported gradually. But different forms of CP reports may prevent the evaluation and spreading of CP. The authors recommend guidelines for evidence-based Chinese medicine CP report, including 18 items which form 5 big sections, by precise item selection and rigorous expert consensus. All these items can embody the principles of Chinese medicine and comply with the requirements of the clinical practice of Chinese medicine. All above are based on synthesizing the published literature systematically and analyzing the current status in this field, also according to the characteristics of Chinese medicine and methodology requirement of evidence-based medicine.

Objective To evaluate the intraoperative opioid?sparing effect of different duration transcutaneous electrical acupoint stimulation ( TEAS ) in video?assisted thoracoscopic lobectomy. Methods Seventy?five patients, aged 18-64 yr, weighing 40-96 kg, of ASA physical status Ⅰ or Ⅱ, scheduled for elective video?assisted thoracoscopic lobectomy under general anesthesia, were randomly divided into 3 groups (n=25 each) using a random number table: control group (group C), 30 min of stimulation before induction of anesthesia group ( group B) , and stimulation throughout surgery ( group T) . In group B, the patients received TEAS ( frequency 2∕100 Hz ) on acupoints Xinshu ( BL15 ) , Feishu (BL13), Neiguan (PC6), Hegu (LI4) on the operated side starting from 30 min before induction of anesthesia until the beginning of induction, and the intensity was the maximum current that could be tolerated. The intensity for Neiguan ( PC6) and Hegu ( LI4) was 6-12 mA, and for Xinshu ( BL15) and Feishu ( BL13 ) was 9-18 mA. In group T, the patients received TEAS on the four acupoints mentioned above starting from 30 min before induction of anesthesia until the end of surgery. The patients had the electrodes applied, but received no stimulation in group C. After anesthesia was induced with propofol?sufentanil?cisatracurium, double lumen endotracheal tube was inserted. Propofol was given by target?controlled infusion to maintain BIS value within the range of 40-60. Cisatracurium was infused continuously to facilitate muscle relaxation. The infusion rate of remifentanil was adjusted to maintain analgesia nociception index value within the range of 50-70. The intraoperative consumption of remifentanil ( the intraoperative consumption of sufentanil was converted to the consumption of remifentanil producing the equivalent effect by 1∶ 10) was recorded. Results Compared with group C, the intraoperative consumption of remifentanil was significantly decreased in B and T groups. The intraoperative consumption of remifentanil was significantly lower in group T than in group B. Conclusion TEAS on Xinshu ( BL15 ) , Feishu (BL13), Neiguan ( PC6) and Hegu acupoints throughout surgery and for 30 min before induction of anesthesia significantly reduces intraoperative opioid consumption in the patients undergoing video?assisted thoracoscopic lobectomy, while TEAS throughout surgery provides better effect.

Objective To evaluate effects of trichostain A (TSA) on cell proliferation, cell cycles, apoptosis and invasiveness of multiple myeloma cell line U266; as well as active changes of methylation regulating proteins including DNA methyl-transferase(DNMTs), methyl-binding domain (MBD) proteins: MBD2 and MeCP2 after treated with TSA. Methods U266 cells were treated with different concentrations of TSA for 12, 24, 48 and 60 h. The proliferation activity of U266 cells was detected by MTT and the IC50 of 24 h was calculated. After U266 cells were treated with IC50, cell cycles were check out by dying with PI. mRNA of matrix metalloproteinase-2(MMP-2), bc1-2, bcl-xl and methylation regulating proteins (DNMTs, MBD2 and MeCP2) were detected by real-time PCR. FCM and Western blotting were used to measure expressions of MMP-2 and MBD2. Results MTT results revealed TSA inhibited proliferation of U266 cells in a dose-and time-dependent manner and the IC50 of 24 h was 0.07 μmol/L FCM analysis showed that TSA could arrest the cell cycle in G0/G1 and the proliferation index (PI) in U266 cells [(49.90 0.39)%]were significantly different after exposed to TSA (0.7 μmnol/L for 24h compared with that in the control cells[(55.78 0.49)%](P <0.01). After treated by TSA, the 2-△△Ct of MMP-2, bcl-2 and bcl-xl were 0.71 0.06, 5.04 0.92 and 2.95 0.35, respectively. There were great changes on mRNA of DNMT, MBD2 and MeCP2. TSA could reverse the transcription of DNMT, MBD2 and MeCP2. Conclusion TSA can arrest the U266 cell cycle in GVG, to prevent its proliferation and promote apoptosis, which maybe greatly connect with the changes of the methylation regulating proteins.