Objective To establish a method for studying molecular mechanism of Rhubarb inhibiting anti-Yersinia pesti based on DNA microarray.Methods A whole genome DN A microarray containing 4005 annotated genes of Yersiniapesti Was used.The minimal inhibitory concentration(MIC)of Rhubarb to Yersiniapestiwas determined by liquid dilution method.The gene expression profile of Yersinia pesti was performed after the exposure to Rhubarb at a concentration of 10×MIC for 30 minutes.The total RNA extracted and purified from Yersinia pesti Was reversely transfected to cDNA and labeled by Cy3-Cy5 dye.The labeled probes were hybridized to the microarray anti the results were obtained by a laser scanner and the microarray data was confirmed by real-time quantitative RT-PCR.Results The platform of the DNA microarray-based bacteria transcriptional profile was established.A total of 498 genes of Yersinia pesti changed significantly in response to Rhubarb.Among them.358 genes were up-regulated,140 down-reguated.Conclusions The whole genome DNA microarray can be used in the studying of molecular anti-Yersinia pesti mechanism of Rhubarb.

OBJECTIVETo explore the effective methods for the correction of lower eyelid retraction at different degree.

METHODS258 patients with lower eyelid retraction were treated in our department since 1999. The lower eyelid retraction could be divided into mild, moderate and severe degree. The lateral canthal anchoring (n = 150), Hamra's lower eyelid blepharoplasty (n = 80) and translid cheek lifting (n = 28) were adopted according to the severity. The therapeutic effect for different degree of lower eyelid retraction was compared.

RESULTS98 patients were followed up for 3-12 months, including 51 patients of mild degree, 29 patients of moderate degree, and 18 patients of severe degree. The retraction were corrected completely in 91 patients. The lower eyelid was repositioned to the level of inferior limbus without inferior scleral show when eyes opened. The palpebral fissure could close completely. The blunt lateral canthus turned to be acute and the scar was inconspicious. The retraction was improved, but not corrected completely in 3 patients of moderate degree and 4 patients of severe degree. The complications included petechiae, chemosis, and so on.

CONCLUSIONSThe lower eyelid retraction can be corrected effectively if the appropriate techniques are performed according to the degree of retraction.

Adult ; Aged ; Blepharoplasty ; methods ; Eyelid Diseases ; surgery ; Eyelids ; surgery ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Treatment Outcome

OBJECTIVETo investigate the feasibility of in vitro proliferation of rat Leydig cells by modifying the cell culture system.

METHODSLeydig cells were isolated from three-week-old rats by a procedure combining collagenase dispersion, stainless steel mesh infiltration and differential adhesion. The isolated cells were cultured in DMEM/F12 and modified media for stem cell proliferation, and the proliferation of the cultured cells was evaluated by cell counting and MTP test. The expression of 3beta-HSD in the cultured cells was detected by immunohistochemistry and flow cytometry, and testosterone productivity in the isolated Leydig cells with or without hCG stimulation was determined at 2 hours and 4 days after cell isolation.

RESULTSThe Leydig cells cultured in the modified media proliferated actively, with a doubling time of (2.26 +/- .31) days, as compared with (16.32 +/- 2.14) days for those cultured in the traditional media (P <0.05). The 3beta-HSD positive rate in the cultured cells was (554.3 +/- 7.1)% after 2 hours and (93.6 +/- 4.6)% after 4 days of culture. All the proliferated cells exhibited testosterone productivity, and their testosterone secretion was significantly upregulated by hCG stimulation (P <0.05).

CONCLUSIONLeydig cells isolated by differential adhesion proliferate actively in the modified culture media.

Animals ; Cell Count ; Cell Culture Techniques ; Cell Proliferation ; Cells, Cultured ; Leydig Cells ; cytology ; Male ; Rats ; Rats, Wistar ; Testosterone ; secretion

OBJECTIVETo analyze the molecular characters of the W135 Neisseria meningitidis strain firstly isolated from patients in Guangdong province.

METHODSBiochemical profile by using the API NH system (bio-Merieux, France) was used for confirmation,and sero-grouping of the meningococcal isolates including one serogroup W135, one serogroup C and three serogroups of a Neisseria meningitidis isolated from patients in Guangdong province in recent two years were performed. The subtype was determined after amplifying porA and porB respectively from the genome DNA of Neisseria meningitidis. Multilocus sequence typing (MLST) was performed for determining the allele profiles and the sequence types (STs). The polygenetic tree was obtained by analyzing the allele profiles with program Splits tree online. The molecular characters of the serogroup W135 Neisseria meningitidis was analyzed by its evolution relationship and the variable regions in porA and porB which encoding the outer membranes proteins (OMPs).

RESULTSThe subtype determined by porA variable regions of the serogroup W135 Neisseria meningitidis was P1.5,2, which was one of the most invasive types. The types of variable regions (VRs) I, IV, V, VII with porB were 1, 1, 1, 17, and there was no VI and VIII in porB. The allele profile of the W135 strain in this study was 2, 123, 4, 3, 8, 4, 6, and its sequence type was ST-2960, which belonged to ST-11/ET-37 clone complex. The subtypes of the serogroup C and serogroup A strains were P1.20, while their types of VR IV were all 7, and they all hadn't VR VII in porB. The strain serogroup C belonged to ST-4821 clone complex, and the 3 serogroup A strains belonged to ST-5 clone complex.

CONCLUSIONThe molecular character of the serogroup W135 Neisseria meningitidis should be the same with the strains isolated in foreign country, and be different from the epidemic types in the area. This serogroup W135 Neisseria meningitis isolated from patients in Guangdong for the first time was thought to be a new type appearing in the local area.

Bacterial Typing Techniques ; China ; epidemiology ; DNA, Bacterial ; Disease Outbreaks ; Encephalomyelitis ; epidemiology ; microbiology ; Genotype ; Humans ; Molecular Sequence Data ; Neisseria meningitidis, Serogroup W-135 ; classification ; genetics ; isolation & purification ; Sequence Analysis, DNA

OBJECTIVE: To explore the underlying mechanism of inhibition by Jinkui Shenqi Pills (JKSQP) on glucocorticoid-enhanced axial length elongation in experimental lens-induced myopia (LIM) guinea pigs. METHODS: Sixty 2-week old male guinea pigs were randomly divided into 4 groups with 15 guinea pigs in each group, according to the random numbers generated by SPSS software: control, LIM, saline and JKSQP groups. The control group includes animals with no treatment, while the guinea pigs in the other 3 groups received lens-induced myopization on the right eyes throughout the experiment (for 8 weeks). The saline and JKSQP groups were given daily intraperitoneal injections of 10 mg/kg hydrocortisone for 2 consecutive weeks at the same time, and then orally administered either saline or JKSQP [13.5 g/(kg•d) for 6 consecutive weeks. Body weight, anal temperature and animal appearance were observed and recorded to evaluate the GC-associated symptoms. The ocular parameters, including refraction and axial length, were measured by streak retinoscopy and A-scan ultrasonography, respectively. The levels of plasma hormones associated with the hypothalamic-pituitary-adrenal axis (HPAA), including free triiodothyronine, free thyroxine, estradiol and testosterone, were measured by radioimmunoassay, and cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate were measured by enzyme-linked immunosorbent assay. In addition, the mRNA and protein expressions of retinal amphiregulin (AREG) was measured by quantitative real-time polymerase chain reaction and Western blotting, respectively. RESULTS: JKSQP effectively increased body weight and anal temperature, improved animal appearance and suppressed axial length elongation in glucocorticoid-enhanced myopic guinea pigs with normalization of 4 HPAA-associated plasma hormones (all P<0.05). The plasma level of cAMP was significantly increased, whereas the plasma level of cGMP and the mRNA and protein expressions of retinal AREG were decreased after treatment with JKSQP (all P<0.05). CONCLUSION JKSQP exhibited a significant inhibitory effect on axial length elongation with decreased expression of AREG in the retina, and normalized 4 HPAA-associated plasma hormones and the expression of cAMP and cGMP in GC-enhanced myopic guinea pigs.
Guinea Pigs ; Male ; Animals ; Glucocorticoids ; Hypothalamo-Hypophyseal System ; Pituitary-Adrenal System ; Myopia/metabolism* ; Body Weight ; RNA, Messenger ; Disease Models, Animal