Purpose The research was conducted to evaluate the stability of N-terminal site-specific PEGylated uricase. Methods The enzyme activity is used as an index to evaluate the thermal stability (at 4-80 ℃) , the pH stability, the ability of avoiding the trypsin digestion and half-life in vivo of PEG-uricase, then it is compared with uricase. Results PEGylated uricase is thermally more stable than uricase between 4 ℃ and 60 ℃, but at 70 ℃, the enzyme activity of both PEG-uricase and uricase decreases sharply. In the test of pH stability, the curve of uricase shows an obvious decrease of enzyme activity at 5 .2-6.0 and 9.2-10.0. The behavior of PEG-uricase indicates that the destabilizing was prevented effectively by PEGylation. The test of anti-trypsin digestion suggests that PEG-uricase retains 70% of its enzyme activity,but uricase only 20% ,200 minutes after being reserved in trypsin solution. Stability in vivo indicates that the half-life of PEG-uricase is 1 530 min and uricase, only 45 min. Conclusion PEGylated uricase has improved thermal stability, the pH stability, ability of protecting from trypsin digestion and stability in vivo.

AIM: To study the effectiveness of hyaluronidase periocular injection combined with α - lipoic acid and Chinese medicine in the treatment of diabetic vitreous hemorrhage.
METHODS: Retrospective analysis of 67 patientswith proliferative diabetic retinophathy ( PDR ) and vitreous hemorrhage, were divided into two groups: treatment group and control group. There were 32 cases in every group, the treatment group were giving hyaluronidase periocular injection combined with α - lipoic acid intravenously and with traditional Chinese medicine in promoting blood circulation to remove blood stasis, control group were only given Chinese medicine treatment in promoting blood circulation to remove blood stasis.
RESULTS: Followed up 2-3mo, the total efficiency of treatment group ( 78%) was better than control group (57%), there was statistically significant difference ( P<0-05 ) . The efficiency of the two groups' patients with vitreous hemorrhage more than 2wk time was 22% and 25% respectively, there was no statistically significant difference in the two groups (P>0. 05).
CONCLUSION: Hyaluronidase periocular injection combined with α-lipoic acid and Chinese medicine in the treatment of PDR combined with vitreous hemorrhageis more effective than only use Chinese medicine. But it only has effect for the early, over 2wk, it has no significant effect.

AIMTo investigate the effects of human urotensin II (hUII) on in vivo pia mater microcirculation in rats.

METHODSAdult SD rats were randomly assigned to the following groups: control, sodium chloride injection (NS), UII(10(-6) mol/L), noradrenaline (NA, 10(-6) mol/L), and UII (10(-6) mol/L) + NA (10(-6) mol/L) groups. For recording of microcirculation images in pia mater, skull windows were performed and mounted on the stage of an intravital microscope equipped with a TV camera. Video images of microcirculation were stored by a video cassette recorder. Temporal changes in internal diameter and microcirculatory velocity of microvessels were measured by computer using the Image Pro software. The blood flow in cerebral tissues were measured with PIMII laser Doppler perfusion Imager (Lisca, Sweden).

RESULTSThe internal diameters of arterioles and venules in control group were (35.4 +/- 3.6) microm and (40.6 +/- 8.5) microm, respectively. In UII group, the arterioles and venules contracted immediately after treated with UII and up to the peak at 1 min, the internal diameters of arterioles and venules were (25.6 +/- 3.4) microm and (23.4 +/- 3.3) microm, respectively (P < 0.05). Both microcirculatory velocity in arterioles and venules had no significant changes in UII group (P > 0.05). The blood flow in meninges increased 1 min after treated with UII and up to high peak at 5 min (3.5 +/- 0.4 perfusion unit vs. control 2.3 +/- 0.6, P < 0.05).

CONCLUSIONhUII can contract microvessels in pia mater of rats and increase microcirculatory blood perfusion to cerebral tissue involved.

Animals ; Cerebrovascular Circulation ; drug effects ; Humans ; Male ; Microcirculation ; drug effects ; Rats ; Rats, Sprague-Dawley ; Urotensins ; pharmacology

The response rules of pressing pain on the back section in the Governor Vessel in patients with gastro-esophageal reflux disease (GERD) were studied to provide references for the diagnosis and treatment of GERD. Seventy-six cases of GERD were included into an observation group while 30 healthy volunteers were recruited into a control group. A mechanical measurement device of pressing pain that could measure the pain threshold was adapted to observe the pressing pain on the back section in the Governor Vessel in GERD patients and healthy volunteers. The test area is from spinous process of the 1st thoracic vertebra to that of the 12th thoracic vertebra (T1 -T12), including acupoints and non-acupoints on the Governor Vessel. As a result, in the observation group the pain threshold of T5-T7 spinous process clearance, which was the location of Shendao (GV 11), Lingtai (GV 10) and Zhiyang (GV 9), was lower than that in the control group (all P < 0.05). This result indicated that there was significant pressing pain in T5-T7 spinous process clearance in patients with GERD, which could be taken as an important auxiliary diagnosis and a new thinking method in the treatment of GERD with acupuncture.
Adolescent ; Adult ; Aged ; Diagnosis, Differential ; Female ; Gastroesophageal Reflux ; diagnosis ; physiopathology ; Humans ; Male ; Meridians ; Middle Aged ; Pressure ; Sensation ; Thoracic Vertebrae ; physiopathology ; Young Adult

Alterations of the autophagy-lysosomal pathway (ALP) and autophagy have been involved in lung ischemia-reperfusion (I/R) injury. However, dynamic imaging of ALP function under lung I/R injury particularly is not fully understood. Here we depicted the live-cell fluorescence imaging of autophagosome to monitor ALP activation and autophagy function. The pAsRed2-N1-LC3 vectors were transfected into CRL-2192 NR8383 (an alveolar macrophage cell line) and CCL149 (an alveolar epithelial cell line) successfully. 0-h, 2-h, 4-h, and 6-h hypoxia/0-h, 2-h, 4-h, and 6-h reoxygenation were then induced with an ALP inhibitor (3-MA) or activator (rapamycin) in the culture of transfected cells separately. ALP activation was conformed by up-regulating AMPK and beclin1 expression. Apoptosis was not obvious in 2-h hypoxia/2-h reoxygenation. pAsRed2-N1-LC3 CCL149 and pAsRed2-N1-LC3 NR8383 cells revealed gradually enhanced AsRed2 from 2-h to 6-h hypoxia/reoxygenation. AsRed2 varied sensitively to 3-MA and rapamycin interventions during 2-h hypoxia/reoxygenation. Our data provides a simple method of autophagosome imaging to monitor ALP activation and autophagy function in lung I/R injury.
Animals ; Autophagy ; Base Sequence ; DNA Primers ; Hypoxia ; physiopathology ; In Vitro Techniques ; Lung ; physiopathology ; Lysosomes ; physiology ; Oxygen Inhalation Therapy ; Rats ; Real-Time Polymerase Chain Reaction

Abstract: Our previous work revealed berberine can significantly enhance the susceptibility of fluconazole against fluconazole-resistant Candida albicans, which suggested that berberine has synergistic antifungal activity with fluconazole. Preliminary SAR of berberine needs to be studied for the possibility of investigating its target and SAR, improving its drug-likeness, and exploring new scaffold. In this work, 13-substitutited benzyl berberine derivatives and N-benzyl isoquinoline analogues were synthesized and characterized by 1H NMR and MS. Their synergetic activity with fluconazole against fluconazole-resistant Candida albicans was evaluated in vitro. The 13-substitutited benzyl berberine derivatives 1a-1e exhibited comparable activity to berberine, which suggested that the introduction of functional groups to C-13 can maintain its activity. The N-benzyl isoquinolines, which were designed as analogues of berberine with its D ring opened, exhibited lower activity than berberine. However, compound 2b, 2c, and 4b showed moderate activity, which indicated that berberine may be deconstructed to new scaffold with synergistic antifungal activity with fluconazole. The results of our research may be helpful to the SAR studies on its other biological activities.
Antifungal Agents ; pharmacology ; Berberine ; pharmacology ; Candida albicans ; drug effects ; Drug Resistance, Fungal ; Drug Synergism ; Fluconazole ; pharmacology ; Isoquinolines ; pharmacology ; Microbial Sensitivity Tests

Objective To investigate the dynamics and development trends of drinking water type of endemic fluorosis after water improvement in Xinbaerhuyouqi of Hulunbeir city, Inner Mongolia and to provide a scientific evidence for the development of countermeasures. Methods We mainly selected Adunchulusumu and Kerlunsumu in Xinbaerhuyouqi of Hulunbeir city as the two monitoring points after water improvement in 2000 -2009. Of these, 1 sample of centralized water supply source water and 3 samples of tap water and 5 samples of noncentralized water supply source water according to water well locations of east, west, south, north and center were collected and the levels of water fluoride were tested; the prevalence of dental fluorosis of school children aged 8 to 12 were examined; from 2002 onwards, the urine samples of 30 children aged 8 to 12(five age groups, six urine samples for each age group) were collected, and all urine samples were collected in the case of less than 30, and urine fluoride was tested. Dental fluorosis was diagnosed using Dean method; water fluoride was tested using fluoride ion selective electrode(WS/T 106-1999); urinary fluoride was tested by determination of fluoride in urine using ion-selective electrode(WS/T 89-1996). Results In 2000 - 2009, the mean levels of fluorine in drinking water in Adunchulusumu and Kerlunsumu were 1.79 - 4.35 mg/L and 1.38 - 3.18 mg/L, respectively; the detection rate of dental fluorosis of children aged 8 to 12 were 45.24％(19/42) - 89.78％(123/137) and 40.00％ (28/70) - 74.47％ (70/94), respectively; the median urinary fluoride of them were 2.30 - 4.15 mg/L and 2.73 - 4.55 mg/L, respectively. ConclusionsThe detection rate of children's dental fluorosis remains high in Xinbaerhuyouqi during the past 10 years after changing water. The endemic fluorosis remains a serious disease. Effective prevention and control measures must be taken to control the occurrence of fluorosis in the future.

Objective To verify the presence of functional subsets of natural killer cells based on the cytokine production. Methods NK cells were purified and cultured in complete RPMI1640 medium in the presence of either IFNγ+anti-IL-4(classical Thl polarization) or IL-4 +anti-IFNγy(classical Th2 polarization) for three days, and then were collected and detected for type Ⅰ/type Ⅱ cytokines by RT-PCR method. Results NK cells were purified from 15 healthy donors, over 70% purity of NK cells were determined by flow cytometry. NK cells in peripheral blood expressed high level of type Ⅰ cytokines, mainly IFNγ, but low level of type Ⅱ cytokines such as IL-10 and IL-13, IL-4 was not produced by NK cells. Cells cultured in IFNγ+ anti-IL-4 condition exhibited significantly increased level of IFNγ, unchanged IL-2, and de creased type Ⅱ cytokines. Cells grew in IL-4 + anti-IFNγcondition exhibited increased IL-10 and IL-13,and decreased IFNγ expressions. Conclusions Based on the cytokine production, NK cells may be divided into two functional subsets in the same manner as that of T lymphocytes(e.g. Th1/Th2): .NKh1 and NKh2. The biological characterization and phenotypic marker are under investigate.

ObjectiveTo investigate the effect of drinking brick tea with low-fluoride level on prevention of tea type fluorosis.MethodsHandahangacha,Hadayinggegacha,Dalainuoyi town,in Keshiketengqi Inner Mongolia endemic fluorosis area were selected as test points,and brick tea with fluoride [(204.5 ± 10.2),(308.2 ±15.4)mg/kg] was given for 12 months.Dental fluorosis,clinical skeletal fluorosis,and X-ray diagnosis of skeletalfluorosis [according to “Endemic Skeletal Fluorosis Diagnostic Criteria” (WS 192-2008)] of adults 20 to 70 years of age were examined and level of fluoride before and after the prevention trial,in brick tea,drinking water,milk tea and urine were tested (fluoride ion selective electrode method),and fluoride intake through tea was calculated.ResultsDetection rate of adult dental fluorosis in Handahangacha was 68.89％ (62/90),clinical detection of skeletal fluorosis was 55.32％ (52/94),and X-ray detection of skeletal fluorosis was 65.17％ (58/89); adult dental fluorosis detection rate in Hadayinggegacha was 54.84％(51/93),clinical detection of skeletal fluorosis was 65.69％(67/102),and X-ray detection rate of skeletal fluorosis was 61.36％ (54/88).Brick tea fluoride was (831.4 ±138.9),(864.3 ± 134.6)mg/kg before the prevention trial in Handahangacha and Hadayinggegacha,respectively,drinking water fluoride content was (0.27 ± 0.05),(0.54 ± 0.24)mg/L,fluoride content of milk tea was (216 ± 1.12),(2.82 ± 1.38)mg/L,adult urine fluoride content was (2.78 ± 1.57),(2.96 ± 1.80)mg/L,and fluoride intake through milk tea was (8.12 ± 5.84),(6.42 ± 5.04)mg/d,respectively; after the prevention trial the fluoride content of brick tea was (204.5 ± 10.2),(308.2 ± 15.4)mg/kg,fluoride content of drinking water (0.34 ± 0.11),(0.62 ± 0.30)mg/L,fluoride content of milk tea(0.97 ± 0.33),(1.83 ± 0.66)mg/L,fluoride content in urine(1.29 ± 0.55),( 1.47 ±0.62)mg/L,fluoride intake through milk tea (3.45 ± 2.05),(3.71 ± 2.07)mg/d,respectively; in Handahan and Hadayinggegacha after the prevention trial the fluoride in brick tea,milk tea,urine fluoride,and fluoride intake through milk tea was significantly lower than that before the trial (t =14.30,12.97 ;6.46,3.95;6.69,5.72;6.27,3.57,all P ＜ 0.01 ).Fluoride intake in Handahangacha through milk tea was within the state heath standard limits( ＜ 3.5mg/d).ConclusionDrinking low-fluoride brick tea can prevent drinking brick tea type fluorosis,the preventive effect is especially more reliable with low fluoride brick tea (204.5 ± 10.2)mg/kg.

OBJECTIVETo explore the quality of life among people living with HIV/AIDS (PLWHA) and analyze the influencing factors in Kunming.

METHODSA cross-sectional survey was conducted from March to April in year 2011, and 247 PLWHA were selected by convenience sampling from Yunnan CDC, Yunnan and Kunming infectious disease hospitals. General questionnaires, the simplified Chinese edition of medical outcomes study-HIV health survey (MOS-HIV, including 11 dimensions) and Social Support Scale were used. t test and multivariable linear regression model were used to analyze the results.

RESULTSFor the subjects investigated, the age was (39.8 +/- 11.9) years old, the median (quartile) value of symptoms related to HIV infection was 1(2). As to the scores of quality of life measured by MOS-HIV, physical summary score was 47.4 +/- 11.2, mental summary score was 43.6 +/- 9.7; for the scores of 11 dimensions of the MOS-HIV,that were general health (42.9 +/- 19.9), physical function (79.4 +/- 24.9), role function (59.8 +/- 48.2), social function (67.0 +/- 33.6), cognitive function (71.0 +/- 25.4), pain (81.3 +/- 26.2), mental health (62.0 +/- 22.3), vitality (49.3 +/- 23.8), health distress (74.4 +/- 21.0), quality of life (51.8 +/- 21.1), health transition (49.0 +/- 29.8). The total score of social support was 28.6 +/- 7.6, of which the score of subjective social support was 17.2 +/- 6.3, the score of the objective social support was 5.9 +/- 2.2; the score of the utilization of social support was 5.5 +/- 1.9. Multivariable linear regression analysis showed that the more the symptoms related to HIV infection, the lower the physical summary scores (standardized coefficients b' = -0.22), the general health (b' = - 0.31), the physical function (b' = -0.16), the role function (b' = -0.23), the pain (b' = -0.21), the mental health (b' = -0.22), the vitality (b' = -0.22) and the health distress scores (b' = - 0.24) (all P values < 0.05); the older the age, the lower the physical summary scores (b' = - 0.16), the mental summary scores (b' = - 0.16), the physical function (b' = -0.26), the vitality (b' = -0.26) and the quality of life scores (b' = -0.17) (all P values < 0.05); the higher the score of the subjective social support,the higher the physical summary scores (b' = 0.26), the mental summary scores (b' = 0.22), the general health (b' = 0.27), the social function (b' = 0.26), the mental health (b' = 0.15) and the quality of life scores (b' = 0.22) (all P values < 0.05); the higher the score of the utilization of social support, the higher the physical function (b' = 0.16) and the health transition scores (b' = 0.31) (all P values < 0.05).

CONCLUSIONPLWHA in Kunming have relatively lower scores of quality of life. A large number of symptoms during infection, older age and lower score of subjective social support were the hazard factors of quality of life in PLWHA.

Acquired Immunodeficiency Syndrome ; epidemiology ; Adult ; China ; epidemiology ; Cross-Sectional Studies ; Factor Analysis, Statistical ; Female ; HIV Infections ; epidemiology ; Humans ; Male ; Middle Aged ; Quality of Life ; Social Support ; Surveys and Questionnaires