Accurate determination of HIV-1 proviral burden and viral load is very useful in prognosis of HIV-1 infected patients and in assessment of drug for therapy of AIDS patients. In order to establish a quantitative method in detecting HIV-1 proviral burden and viral load, 8E5 cell line and a recombinant RNA constructs were used as the HIV-1 proviral DNA and viral RNA external references, respectively. The PCR products were labeled with the fluorescent DNA dye SYBR green. The amount of burden or load was measured by GeneAmp 5700 Sequence Detection System. Using this method, the HIV-1 proviral burdens in PBMC of patient and in cell suspension treated with the compounds AZT, GL and WT were measured. HIV-1 viral loads in supernatant of the cell culture treated with the above compounds were also determined. The therapeutic indices (TIs) of the compounds calculated based on the inhibition of virus induced syncytial formation, and inhibitionn of proviral burdens and viral loads were compared, and their TIs successively increased. The fluorescent real time quantitative PCR possesses very good specificity, sensitivity and duplication. TI value of a drug based on inhibition of proviral burden in cell culture, and the proviral burden in PBMC of patient may be useful in evaluating a drug on eradicating provirus from resting and memory CD4 T cells.

AIMTo demonstrate the specific killing of folate receptor (FR)-positive tumor cells can be achieved by folate-targeted penicillin-G amidase (PGA) combined with its prodrug substrate N-(phenylacetyl) doxorubicin (DOXP).

METHODSFolic acid was covalently linked to PGA and folate content value was determined by quantitative UV spectrophotometry. The ability of folate conjugated PGA to hydrolyze DOXP was measured by RP-HPLC. Visual demonstration of uptake by FR (+) HeLa and SKOV3 cells was detected by using FITC labeled folate-PGA and a fluorescence microscopy. The cytotoxicity of DOXP towards the cells in the presence or absence of folate-PGA was assayed by using MTT method.

RESULTSThe folate-PGA has a specific activity of 29. 8 U x mg(-1) (protein). FR selectivity was confirmed by fluorescence microscopy. The combination of DOXP prodrug with folate-PGA generated higher cytotoxicity towards the FR (+) cells than free doxorubicin. The IC50 was 0.72 micromol x L(-1) for HeLa cells and 0.75 micromol x L(-1) for SKOV3 cells, respectively. Further, the enhanced cytotoxicity reduced greatly with the addition of free folic acid.

CONCLUSIONFolate conjugated PGA did not significantly compromise PGA catalytic activity and enabled binding prodrug-activating enzyme PGA to folate receptor expressing cells, and increased the sensitivity of the cells to doxorubicin followed by administration of its prodrug substrate.

Antibiotics, Antineoplastic ; pharmacology ; Carrier Proteins ; metabolism ; Cell Line, Tumor ; Doxorubicin ; analogs & derivatives ; pharmacology ; Drug Delivery Systems ; Female ; Folate Receptors, GPI-Anchored ; Folic Acid ; chemistry ; pharmacology ; HeLa Cells ; Humans ; Inhibitory Concentration 50 ; Ovarian Neoplasms ; pathology ; Penicillin Amidase ; chemistry ; pharmacology ; Prodrugs ; pharmacology ; Receptors, Cell Surface ; metabolism

OBJECTIVETo establish a multiplex RT-PCR-based reverse dot blot hybridization technique to detect influenza viruses.

METHODSObtain the HA nucleotide sequences of seasonal influenza H1N1, seasonal influenza H3N2, influenza H1N1 and human avian influenza H5N1 from GenBank. Design primers in conservative district and probes t in high variable region respectively, after analyzing the HA nucleotide sequences of influenza virus through the Vector NTI 9.0. Establish and optimize multiple RT-PCR system by comparing amplification efficiency and specificity at different primer concentrations. Establish the reverse dot hybridization system after optimizing the concentration of probes. To compare the sensitivity and specificity of this technique and the general RT-PCR Method through extracting the viral RNA of the mentioned influenza virus which are to be the reference substance.

RESULTSSuccessfully establish a multiplex RT-PCR-based reverse dot blot hybridization technique for detecting influenza viruses. This technique is 100-1000 times more sensitive than gel electrophoresis method, and it has a good specificity.

CONCLUSIONSuccessfully established multiplex RT-PCR-based reverse dot blot hybridization technique for detecting influenza viruses.

Humans ; Influenza A Virus, H1N1 Subtype ; genetics ; isolation & purification ; Influenza A Virus, H3N2 Subtype ; genetics ; isolation & purification ; Influenza A Virus, H5N1 Subtype ; genetics ; isolation & purification ; Influenza, Human ; diagnosis ; virology ; Nucleic Acid Hybridization ; methods ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Sensitivity and Specificity

OBJECTIVETo investigate the expression and the localization of Cathepsin K and IL-6 mRNA in root-resorbing tissue and to elucidate the molecular changes and mechanism of root resorption induced by tooth movement.

METHODSRats were subject to experimental tooth movement to induce root resorption. In situ hybridization was performed to identify the cells in root-resorbing tissue that produced Cathepsin K or IL-6 the difference of CK mRNA or IL-6 mRNA expression between root resorption group and control group was calculated by t-test.

RESULTSCathepsin K mRNA was highly and selectively expressed in multinuclear odontoclast and IL-6 mRNA expressed in fibroblast, osteoblast, osteocyte and cementoblast. The expression of Cathepsin K mRNA and IL-6 mRNA in root-resorbing tissue increased evidently compared with the normal periodontium.

CONCLUSIONSOdontoclast in the root-resorbing tissue expresses Cathepsin K mRNA that participates in proteolysis during root resorption. IL-6 plays a very important role in the root resorption as a multifunctional cytokine.

Animals ; Cathepsin K ; Cathepsins ; biosynthesis ; genetics ; Interleukin-6 ; biosynthesis ; genetics ; Male ; Osteoclasts ; enzymology ; RNA, Messenger ; biosynthesis ; genetics ; Rats ; Rats, Wistar ; Tooth Movement Techniques ; Tooth Resorption ; enzymology

The purpose of this study was to evaluate whether the cranial and circumaxillary sutures react differently to maxillary expansion (ME) and alternate maxillary expansions and constrictions (Alt-MEC) in a rat model. Twenty-two male Sprague-Dawley rats (6 weeks old) were used and divided into three groups. In ME group (n=9), an expander was activated for 5 days. In Alt-MEC group (9 animals), an alternate expansion and constriction protocol (5-day expansion and 5-day constriction for one cycle) was conducted for 2.5 cycles (25 days total). The control group comprised 4 animals with no appliances used, each of two sacrificed on day 5 and day 25 respectively. Midpalatal suture expansion or constriction levels were assessed qualitatively and quantitatively by bite-wing X-rays and cast models. Distances between two central incisors and two maxillary first molars were measured on cast models after each activation. Circumaxillary sutures (midpalatal, maxillopalatine, premaxillary, zygomaticotemporal and frontonasal suture) in each group were characterized histologically. Results showed that midpalatal suture was widened and restored after each expansion and constriction. At the end of activation, the widths between both central incisors and first molars in Alt-MEC group were significantly larger than those in ME group (P<0.05). Histologically, all five circumaxillary sutures studied were widened in multiple zones in Alt-MEC group. However, only midpalatal suture was expanded with cellular fibrous tissue filling in ME group. Significant osteoclast hyperplasia was observed in all circumaxillary sutures after alternate expansions and constrictions, but osteoclast count increase was only observed in midpalatal suture in ME group. These results suggested that cranial and circumaxillary sutures were actively reconstructed after Alt-MEC, while only midpalatal suture had active reaction after ME.
Animals ; Male ; Mandible ; anatomy & histology ; physiology ; Masticatory Muscles ; anatomy & histology ; physiology ; Maxilla ; anatomy & histology ; physiology ; Range of Motion, Articular ; physiology ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo select the premium alert threshold for major communicable disease by using the control graph alert technique based on the local disease information.

METHOD8 major communicable diseases in Songjiang district were ascertained by analysis of the national early warning detection information system which include the other diarrhea, mumps, chickenpox, scarlet fever, rubella, hand foot mouth disease, influenza and dysentery; weekly reported cases from 2008 to 2011 were used to establish the early detection model (PERCENTILE (array, x), array (4×5), x = 0.05, 0.10…0.95) by moving percentile method, next applying the established early detection model and the golden standard (AKX(-)D ± 2s) to predict the expected weekly cases in 2012 respectively, and then ascertain the predict results by comparison with the actual weekly cases in 2012 respectively, finally the premium threshold was selected by comparison of the model predicted results with the golden standard predicted results after comprehensive consideration of the sensitivity, specificity, positive predictive value and negative predictive value and receiver operating characteristic (ROC) curve.

RESULTSThe premium alert threshold for mumps, other diarrhea and rubella was P90, dysentery was P75, scarlet fever and chickenpox was P80, and the premium threshold for hand-foot-mouth disease (HFMD) and influenza was P95, the sensitivity of 8 major communicable diseases were 100%, 100%, 86%, 100%, 100%, 100%, 94%, 100%, respectively; the specificities were 92%, 73%, 72%, 77%, 73%, 92%, 66%, 80%, respectively; the positive predictive values were 43%, 40%, 32%, 8%, 24%, 20%, 59%, 47%, respectively; and the negative predictive values were 100%, 100%, 97%, 100%, 100%, 100%, 96%, 100%, respectively. The national recommended alert thresholds for the 8 major communicable diseases were P80, except for chickenpox (P50) and HFMD (CUSUM).

CONCLUSION6 out of 8 major communicable diseases' early detection thresholds in Songjiang district should be adjusted according to the analysis results. Premium alert threshold selection need to consider the local disease report and the characteristics of infectious diseases to upgrade the early detection capability.

China ; epidemiology ; Communicable Disease Control ; methods ; Communicable Diseases ; epidemiology ; Disease Notification ; Disease Outbreaks ; prevention & control ; Humans ; Models, Theoretical ; Population Surveillance ; methods

Objective To investigate the epidemiological condition of the sub-health status of the young and middle-aged persons in urban and rural areas of Hubei province, and to explore the relationships among personality, quality of sleep and sub-health status. Methods Purposive sampling method and Personal Health Status Questionnaire developed by Chinese Academy of Medical Sciences were used to investigate the sub-healthy status of 3187 subjects aged 18-55 and evaluate the relationships among sub-health status, personality and the quality of sleep with structural equation model. Results Gender, education, marriage, age and occupation were factors influencing the status of sub-health. Among all of the subjects, sub-health status of females were more serious than that of males (P＜0.0001 of all). The divorced, separated and widowed ones were more likely to experience sub-health than other status of marriage (P= 0.0006, P=0.0102). The sub-health status of the subjects aged 30-55 in the urban (P=0.0197)and low-literacy residents in rural (P=0.0003)were among the worst. The condition of the sub-health in migrant workers and professional were worse compared to other occupational groups (P=0.0043). Structural equation model analysis revealed that sub-health status associated with the quality of sleep and the type of characteristics (P＜0.01 ). Conclusion Sub-health status did exist among the young and middle-aged persons and was influenced by the quality of sleep and personality. However, it could be adjusted by improving the quality of sleep and personality to prevent sub-health.

Taking Guangdong Province Traditional Chinese Medical Hospital as an example,the paper introduces the study idea of hospital IT intelligent operation maintenance and monitoring platform,expounds on design and realization of the platform,including general technical architecture,functional architecture and core business process,summarizes related practical experiences,and points out that application of the platform is able to enhance operation maintenance level and user satisfaction significantly.

OBJECTIVETo investigate the anatomy of penis and its adjacent organ for phalloplasty.

METHODSAnatomic dissection of penis and perineum was performed on 30 adult male cadavers (60 sides). Observation and measurement were focused on the penile length of different parts, the morphological relationship of infundibular ligament and suspensory ligament with penile radix, and the feature of crus penis with relation to the deep penile artery.

RESULTSThe average length of the penile shaft was 8.13 cm, the penile radix was 7.67 cm and the crus penis was 5.96 - 5.98 cm. The deep penile artery penetrated into the crus penis at its middle 1/3. The infundibular ligament attached to superficial fascia of the penis and extended downward to the scrotal septum to constitute the suspensory structure for both of them. The suspensory ligament attached to the dorsal deep fascia of the penis. Becoming thicker, the rear part of the suspensory ligament connected firmly to the pubic arcuate ligament to constitute a part of suspensory mechanism for the urethra. There was a part of cavernous body, which was free from either ligament or bony attachment, between the penile radix and the crus penis, where the dorsal artery and nerve of penis turned around from the ventral to the dorsal aspect of the penis and the penile dorsal vain penetrated the urogenital septum, draining into intrapelvic venous plexus.

CONCLUSIONSThe divisional measurement of the penis length, the recognition of the suspensory ligaments and the anatomic feature of the crus penis with relation to the deep penile artery are all of significant importance to improve the operation of phalloplasty.

Adolescent ; Adult ; Aged ; Humans ; Male ; Middle Aged ; Penis ; anatomy & histology ; Perineum ; anatomy & histology ; Reconstructive Surgical Procedures ; methods ; Young Adult

OBJECTIVETo isolate the bioflocculant-producing bacteria from activated sludge and investigate the flocculating characteristics of the newly isolated bioflocculant.

METHODSBacteria were screened from activated sludge samples to isolate bioflocculant-producing bacteria. Flocculating activity was used as a measure of the flocculating capability of the bioflocculant.

RESULTSA novel bioflocculant-producing bacterium was isolated, which was identified to belong to genus Aeromonas and named as Aeromonas sp. N11. Flocculating activity increased in the presence of K+, Na+, or Ca2+. The highest flocculating activities for kaolin suspension were obtained in acidic pH ranges, and optimum pHs for it were 3.0, 4.0, and 5.0 with 1 mmol/L K+, Ca+, and Na+ present, respectively. The highest flocculating activities for soil suspension were observed at pH 8.0. The bioflocculant had a good flocculating activity and could achieve a flocculating activity of 92.4% for kaolin suspension at a dosage of only 1 mgxL(-1), and its activity in kaolin suspension was decreased by only 9.2% after heating at 100 degrees C for 60 min.

CONCLUSIONThe bioflocculant produced by Aeromonas sp. N11 has strong flocculating activity and high stability, which affords high possibility of its practical use.

Aeromonas ; metabolism ; Culture Media ; Flocculation ; Hydrogen-Ion Concentration ; Kaolin