Background Acanthamoeba keratitis is a sort of serious infectious eye disease with high causing-blindness rate.Acanthamoeba keratitis often is misdiagnosed as fungal keratitis or viral keratitis in the early stage.Because conventional clinical diagnosis methods show a low specificity and take a long time,timely treatment often is delayed.Conventional PCR does not apply well because the lesion sample is not enough to extract DNA.However,direct PCR can amplify 18S rRNA conserved sequence of acanthamoeba keratitis without the extraction of DNA.Objective This study was to discuss the feasibility for rapid diagnosis of acanthamoeba keratitis using direct PCR to amplify the gene 18S rRNA fragment.Methods Ten acanthamoeba strains were isolated from 10 eyes with acanthamoeba keratitis in Qingdao Eye Hospital.The sensitivity of the direct PCR assay was tested using different numbers of amoebas.The specificity of the assay was tested using DNA extracted from acanthamoeba,candida albicans,pseudomonas aeruginosa,herpes simplex virus-1 (HSV-1) and normal human corneal epithelial cell.Acanthamoeba keratitis models were established using infected method in clean 6-week-old female BALB/c mice.Corneal lesion samples were obtained 1 day,3,5,7,10,15 days after modeled.The effectivity and feasibility of the direct PCR assay for rapid diagnosis of acanthamoeba keratitis were evaluated and compared with culture method,corneal smear examination and real-time PCR.Results Direct PCR primers could only amplify DNA of acanthamoeba rather than other pathogens,and 10 stains of acanthamoeba were detected at least in each sample.During the development of acanthamoeba keratitis in the mice,the diagnosis positive rate of direct PCR was 80.0％,90.0％,80.0％,70.0％,70.0％ and 50.0％ in 1 day,3,5,7,10,15 days after modeled with the total positive rate 73.3％,which was higher than 31.7％ of culture method,56.7％ of corneal smear examination and 61.7％ of realtime PCR,with a significant difference between the direct PCR and culture method (P =0.005),but no significant difference was seen in the total positive rate between the direct PCR and real-time PC R (P =0.172) or corneal smear examination (P =0.056).Conclusions The direct PCR assay is a simple,rapid,highly specific and sensitive method for the rapid diagnosis of acanthamoeba keratitis,especially for the limited lesion sample.

OBJECTIVETo improve the rapid propagation system of Pinellia ternata.

METHODDocumentaries at home and abroad were consulted.

RESULTThe tissue culture's conditions and research results of P. ternata were summarized.

CONCLUSIONAdvanced research on the structure of calli and litter tubercle should be made. Traditional rapid propagation system may be improved from the following aspects: the best cooperation of hormones, the most active ex-plant, one time culture and suitable transplant.

Culture Media ; Culture Techniques ; Lighting ; Pinellia ; growth & development ; Plant Growth Regulators ; pharmacology ; Plants, Medicinal ; growth & development ; Temperature

This study was designed to evaluate the effects of drilling through the growth plate and using adipose-derived stem cells (ADSCs) and bone morphogenetic protein-2 (BMP-2) to treat femoral head epiphyseal ischemic necrosis,which can be done in juvenile rabbits.Passage-four bromodeoxyuridine (BrdU)-labeled ADSCs were cultured,assayed with MTT to determine their viability and stained with alizarin red dye to determine their osteogenic ability.Two-month-old,healthy male rabbits (1.2 to 1.4 kg,n=45) underwent ischemic induction and were randomly divided into five groups (group A:animal model control;group B:drilling;group C:drilling & ADSCs;group D:drilling & BMP-2;and group E:drilling & ADSCs & BMP-2).Magnetic resonance imaging (MRI),X-ray imaging,hematoxylin and eosin staining and BrdU immunofluorescence detection were applied 4,6 and 10 weeks after treatment.Approximately 90％ of the ADSCs were labeled with BrdU and showed good viability and osteogenic ability.Similar results were observed in the rabbits in groups C and E at weeks 6 and 10.The animals of groups C and E demonstrated normal hip structure and improved femoral epiphyseal quotients and trabecular areas compared with those of the groups A and B (P＜0.01).Group D demonstrated improved femoral epiphyseal quotients and trabecular areas compared with those of groups A and B (P＜0.05).In summary,drilling through the growth plate combined with ADSC and BMP-2 treatments induced new bone formation and protected the femoral head epiphysis from collapsing in a juvenile rabbit model of femoral head epiphyseal ischemic necrosis.

OBJECTIVETo study the human immunodeficiency virus (HIV) status through heterosexual transmission in Yining city and to provide information on effective intervention measures.

METHODSCohort of HIV sero-discordant couples identified from 1997 to 2000 was formed. Proportional risk model was used to analyze the time of HIV sero-conversion and the related factors. All the recruiters were under informed consent.

RESULTSThrough following on 22 sero-discordant couples, we found that the incidence density (ID) of HIV sero-conversion was 32.49/100 person-year (PY) with 33.74/100 PY for women. In the proportional hazard model, the course of sero-conversion was only 2.43 years and the frequency of sexual contact was statistically significant (>or= 3 times/week vs. < 3 time/week: RR = 1.984, 95% CI: 1.045 - 3.767), indicating this factor was related to the hazard of HIV sero-conversion. However, the viral load of HIV infections has no such effect on HIV sero-conversion of their spouses. In addition, the ratio of CD4(+)/CD8(+) was lower in spouses of HIV sero-conversion than that in spouses of HIV non-sero conversion (t test: t = 4.77, P < 0.01).

CONCLUSIONIn order to control HIV transmission among general population, we suggested that HIV/AIDS counseling and testing be developed for pre-marital people in the region with high HIV prevalence.

Blotting, Western ; China ; epidemiology ; Enzyme-Linked Immunosorbent Assay ; Female ; HIV ; immunology ; HIV Infections ; epidemiology ; immunology ; Humans ; Incidence ; Male ; Proportional Hazards Models ; Sexual Behavior ; Sexually Transmitted Diseases, Viral ; epidemiology ; immunology

OBJECTIVETo explore the central mechanism of moxibustion on analgesic effect.

METHODSMale Wistar rats were screened by pain threshold value before making model, and 48 rats whose pain threshold was (250 +/- 25) g were selected. Twelve male Wistar rats were randomly selected as a normal group. For the rest rats the rheumatoid arthritis (RA) model was duplicated by raising in a windy, cold and wet environment combined with injection of Freund's complete adjuvant (FCA), and then they were randomly divided into a model group, a moxibustion group and a moxa volatile oil group, 12 rats in each group. The moxibustion and the moxa volatile oil igroup were treated with moxibustion and moxa volatile oil at "Shenshu"(BL 23) and "Zusanli"(ST 36), respectively, for 15 days. No interventions were added on the model group and the normal group. The pain threshold in Iinjured foot and the expression of hypothalamic POMC mRNA and PDYN mRNA in rats were observed.

RESULTSCompared with the normal group, the pain threshold and the expression of hypothalamic POMC mRNA and PDYN mRNA in the model group were increased (all P < 0.01). Compared with the model group, the pain threshold and the expression of hypothalamic POMC mRNA and PDYN mRNA in the moxibustion group were increased significantly (all P < 0.01), but no statistically significance in the moxa volatile oil group (P > 0.05). Compared with the moxa volatile oil group, the above-mentioned observative indices in moxibustion group were all increased significantly (all P < 0.01).

CONCLUSIONMoxibustion has obvious analgesic effect and its mechanism may be related to the increasing expression of hypothalamic POMC and PDYN mRNA through the warming effect of moxibustion.

Animals ; Arthritis, Rheumatoid ; genetics ; metabolism ; therapy ; Enkephalins ; genetics ; metabolism ; Humans ; Hypothalamus ; metabolism ; Male ; Moxibustion ; Pro-Opiomelanocortin ; genetics ; metabolism ; Protein Precursors ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Wistar

A new viral sampling concentration device was designed which was equipped with a new cationic filter membrane-Nanoceram suitable for field sampling. Norovirus Genegroup II was detected from environmental water with the aid of this device. The effects on virus recovery of prefiltration, various second-concentration methods, and different eluants were investigated through pre-experiment. The concentration optimized process, and the optimal concentration process were then determined. The results showed that the prefiltration had a profound effect on virus recovery, and two second-concentration method: PEG-NaC1 precipitation and celite adsorption, had almost the same concentration effects. The Na2 HPO4 solution of 0.15 mol/L was selected as the final eluant to elute the adsorbed Nuorovirus from the celite. The virus recovery of Nanoceram was determined to be 3.02%. Finally, successful detection of Norovirus GII in sewage from Yangqiao River, Fengtai District, Beijing was acheived. All these data had shown that the Naneceram filter concentration method could concentrate Norovirus from environmental water with a steady effects.
Filtration ; instrumentation ; methods ; Fractional Precipitation ; instrumentation ; methods ; Genotype ; Norovirus ; classification ; genetics ; isolation & purification ; Rivers ; virology ; Water Microbiology

Objective To study the effects of Jianpi Xiaoai Prescription on cell cycle and apoptosis of human colon cancer HCT116 cells and related factors of Wnt/β-catenin signaling pathway; To investigate its mechanisms of anti-metastasis and recurrence of colorectal cancer. Methods The logarithmic growth phase HCT116 cells were divided into blank group, saline group, 5-Fu group, and Jianpi Xiaoai Prescription low-, medium-, and high-dose groups. After intervention for 48 h, the cells were harvested, and the cell cycle and apoptosis were detected by flow cytometry. The protein expression of β-catenin in the nucleus was detected by Western blot, and the expression of c-myc and cyclinD1 mRNA was detected by PCR. Results Compared with the blank group and saline group, the ratio of HCT116 cells apoptosis of Jianpi Xiaoai Prescription low-, medium-, and high-dose groups increased; the proportion of cells in phase G1 increased and the proportion of S cells decreased; the expression of β-catenin protein in the nucleus and the expression of c-myc,cyclinD1 mRNA decreased,especially in the Jianpi Xiaoai Prescription high-dose group,with statistical significance(P<0.05,P<0.01).Conclusion Jianpi Xiaoai Prescription can promote apoptosis of human colon cancer HCT116 cells and block the cell cycle, and its mechanism may be related to regulating Wnt/β-catenin signaling pathway.

OBJECTIVETo observe the prevalence, blood pressure change in prehypertensive population and associated cardiovascular risk factors.

METHODSData from a prehypertensive cohort defined with the JNC-7 prehypertension diagnostic criteria were obtained in the employees of kailuan group during the health examination between 2006 to 2007 and the same population was revisited between 2008 to 2009 to observe the change of blood pressure and the associated determinants for blood pressure change.

RESULTS(1) There were 25 474 prehypertensive during the 1(st) visit and 8361 subjects developed hypertension during the 2(nd) visit (35.3% in men and 23.3% in women, 27.2% with baseline blood pressure 120 - 129/80 - 84 mm Hg (1 mm Hg = 0.133 kPa) and 43.8% with baseline blood pressure 130 - 139/85 - 89 mm Hg, 34.3% with risk factors and 19.9% without risk factors). (2) Multiple logistic regression analysis showed that the baseline SBP, waist circumference, age, BMI, gender (male), DBP, TC, FBG, TG, LDL-C were the risk factors of blood pressure progression with a RR (95%CI) of 1.052 (1.048 - 1.056), 1.009 (1.006 - 1.013), 1.023 (1.021 - 1.026), 1.063 (1.052 - 1.074), 1.554 (1.442 - 1.675), 1.036 (1.029 - 1.043), 1.064 (1.037 - 1.093), 1.043 (1.024 - 1.062), 1.041 (1.021 - 1.062) and 1.035 (1.000 - 1.072), respectively.

CONCLUSIONA third (32.8%) prehypertensive population progressed into hypertension after two years, baseline SBP, waist circumference, age, BMI, gender (male), DBP, TC, FBG, TG, LDL-C were the risk factors of predicting blood pressure progression.

Adult ; Blood Glucose ; metabolism ; Blood Pressure ; Cardiovascular Diseases ; epidemiology ; Cholesterol ; blood ; Female ; Follow-Up Studies ; Humans ; Hypertension ; epidemiology ; Male ; Middle Aged ; Prehypertension ; epidemiology ; Prevalence ; Risk Factors ; Waist Circumference

OBJECTIVETo sequence the complete sequence of bocavirus I with sequence independent single primer amplification (SISPA-PCR).

METHODSTo exclude the co-effection samples, all clinical samples of diarrhea cases were screened with special primers of rotavirus, astrovirus, adenovirus, calicivirus and bocavirus I. The virus were enriched through ultracentrifugation. Other nucleic acids, such as human and bacteria genomes, were degradated by DNase I and RNase. DNA of bocavirus was Amplificated with SISPA-PCR, then purificated, cloned and sequenced. The sequences were alighmented in nr with blastn and assembled with DNAstar.

RESULTSA 4834bp sequence of bocavirus I were assembled.

CONCLUSIONSISPA-PCR is an economical and efficient technique for sequence a virus complete genome.

Base Sequence ; Bocavirus ; genetics ; isolation & purification ; DNA Primers ; genetics ; Diarrhea ; virology ; Genome, Viral ; Humans ; Parvoviridae Infections ; virology ; Polymerase Chain Reaction

OBJECTIVETo investigate the feasibility of multi-modalities in the reconstruction of circumferential defects after resection of cancers in pharyngoesophageal regions, and to compare the pros and cons between different surgical procedures.

METHODSAccording to the nature and extend of defects, five different methods including pectoralis major myocutaneous flap, laryngeal tube replacement, free jejunum, free forearm flap and gastric pull-up were used to reconstruct the circumferential pharyngoesophageal defects in 72 patients. Function of deglutition and restoration of swallowing was regularly followed up and objectively evaluated.

RESULTSA total of 35 cases of pectoralis major myocutaneous (PM) flaps, 8 cases of laryngeal tube replacement, 12 cases of free jejunum, 12 cases of free forearm flaps and 16 cases of gastric pull-up were performed. Different complications including wound infection, pharyngeal fistula, partial necrosis of PM flap, partial necrosis of gastric wall, stricture of anastomotic site were encountered in 15 cases. All patients survived the operation except one due to partial necrosis of the gastric wall. Two of 4 patients who developed anastomotic stricture can ingest half-liquid food, the remaining cases regained normal deglutition function. The mean postoperative follow-up time was 1. 6 years with 2-year survival rate of 45.3%.

CONCLUSIONSCircumferential defects resulting from resection of carcinomas in pharyngoesophageal region can be reconstructed with different operative techniques depending on the nature and extend of the defects. Once the operative indications are properly selected, the good reconstructive results are to be achieved.

Adult ; Aged ; Esophageal Neoplasms ; surgery ; Esophagus ; surgery ; Feasibility Studies ; Female ; Humans ; Hypopharyngeal Neoplasms ; surgery ; Male ; Middle Aged ; Reconstructive Surgical Procedures ; methods