1.Determination of n-Octanol/Water Partition Coefficients of Naphthalenes and Anthraquinones Using Quantitative Structure-Retention Relationship Models by Reversed Phase-High Performance Liquid Chromatography
Huimin YU ; Shuying HAN ; Haishan DENG ; Yumei CHI
Chinese Journal of Analytical Chemistry 2016;44(6):958-964
n-Octanol/ water partition coefficients (Kow ) is an important parameter commonly used to explain toxicity, activity and transmembrane of drugs. However, it is difficult to be detected by direct experimental determination. In this work, a set of 29 neutral and acidic analogues of naphthalene and anthraquinone with reliable experimental Kow data was chosen as model compounds for establishing linear relationship between the logarithm of apparent n-octanol/ water partition coefficient (lgKow), and the logarithm of reversed phase-high performance liquid chromatography (RP-HPLC) retention factor of the solutes corresponding to neat aqueous fraction of mobile phase (lgkw ) as the quantitative structure-retention relationship (QSRR) model. Methanol-water mixture was used as mobile phase at various pH, and retention time (tR ) was rectified by a dual-point retention time correction (DP-RTC) in this method. The experiment results indicated that the proposed QSRR model had good correlation coefficient R2 = 0. 974 -0. 976 with satisfactory results of internal and external validation (the cross-validated correlation coefficient R2cv of 0. 970-0. 973, and 1. 4% ≤relative error (RE)≤7. 9% for all the 6 verification compounds). In addition, this QSRR model was compared with linear solvation energy relationship ( LSER) involved in different descriptors of molecular structure, showing no differences. The QSRR model was applied to measure Kow of 11 naphthalenes and anthraquinones, and the predicted data were compared with Shake-flask method (SFM) experimental ones, as well as calculated ones obtained by software. The results suggested that the proposed method for Kow determination in this work was more accurate, simple and fast. To the best of our knowledge, this is the first report on measuring Kow data for these compounds. The proposed strategy provides the possibility in determining Kow of lipophilic components in complex mixture more quickly and accurately by RP-HPLC.
2.Study on positive rate of blocking antibody in women with recurrent spontaneous abortion administered by route and frequency of paternal lymphocyte immunotherapy
Hongling YU ; Xiaohui DENG ; Lan CHAO ; Chao CHEN ; Yilong HAN
Chinese Journal of Obstetrics and Gynecology 2013;48(12):903-906
Objective To investigates factors affecting the positive rate of blocking antibody treated by paternal lymphocyte immunotherapy in patients with recurrent spontaneous abortion (RSA).Methods From January 2008 to August 2012,326 RSA cases undergoing treatment in Infertility Center of Qilu Hospital were studied retrospectively.Those patients were divided into 2 groups randomly:260 cases in intradermal injection group were administered via bilateral forearm intradermal injections for immunotherapy once 21 days,then the blocking antibody was determined after 2 (23 cases),3 (73 cases),4 (74 cases),5(90 cases) times respectively,while in subcutaneous injection group,the 66 cases were administered via subcutaneous injection once 21 days,the blocking antibody measured after 3 times; In both cases,the blocking antibody was all determined 2 weeks later.The positive rate of blocking antibodies and the rate of successful pregnancy was recorded,and then followed up after the blocking antibody turning positive.Results (1)Positive rate of blocking antibodies:the positive rate of blocking antibodies were 17% (4/23),58% (42/73),72% (53/74) and 84% (76/90) in the 2,3,4,and 5 times of intradermal injection group,respectively (P < 0.05).In subcutaneous injection group,the positive rate of blocking antibodies was 38 % (25/66),which was significantly lower than that in group intradermal injection receiving 3 times immunotherapy (P <0.05).(2) The rate of pregnancy:the 176 patients out of 200 patients were pregnant when antibody was positive after immunotherapy,with 71.6% (126/176)of patients gained successful pregnancy(the length of pregnancy more than 5 months).Conclusions The route and frequency of administration of immunotherapy could influence the positive rate of blocking antibody.The rate of successful pregnancy will be increased after blocking antibody turning positive.
3.Effect on the expression of inflammatory cytokines ,NF-κB p65 in the brain and serum of preeclampsia model rats by magnesium sulfate
Guilin QI ; Xiaoqing LUO ; Jie DENG ; Han YU
The Journal of Practical Medicine 2017;33(9):1423-1426
Objective To investigate the effect of magnesium sulfate on NF-κB p65,inflammatory cyto-kines in brain and serum of preeclampsia model rats by magnesium sulfate. Methods The healthy SD rats were randomly divided into 4 groups,normal pregnancy group(n = 10),magnesium sulfate control group(n = 10), preeclampsia(PE)model group(n = 10)and magnesium sulfate intervention group(n = 10). Blood pressure, urinary protein,blood urea nitrogen and alanine aminotransferase levels were detected in rats in each group. RT-qP-CR and ELISA were used to detect the expression of IL-1β,TNF-αand IL-6 in the brain and serum. Western blot assay was used to detect the expression of the NF-κB p65 in brain. Results The level of proteinuria,blood urea nitrogen and blood pressure in PE model rats were significantly higher than those in the normal pregnancy group and those in the magnesium sulfate intervention group at 20 days of gestation(P<0.05). Results of RT-qPCR and ELISA assay showed that IL-1β,TNF-α and IL-6 level in magnesium sulfate intervention group were sig-nificantly lower than those in PE model group(P<0.05),but still higher than those in the normal pregnancy group (P < 0.05). Western blot result showed that NF-κB p65 in the magnesium sulfate intervention group was lower than that in the PE group(P<0.05),but still higher than that in the normal pregnancy group(P<0.05). Con-clusion The prevention and treatment preeclampsia mechanism of magnesium sulfate may inhibit inflammatory cyto-kines through NF-κB p65 pathway in the preeclampsia model rats.
4.Homology analysis of clinically isolated and colonized linezolid-resistant Enterococcus faecalis strains from a patient
Zhangya PU ; Zhijian YU ; Zhong CHEN ; Xiangbin DENG ; Bing BAI ; Duoyun LI ; Xiaojun LIU ; Xueying HAN ; Fojun LIN ; Qiwen DENG
Chinese Journal of Infection Control 2017;16(4):343-345,350
Objective To study the homology characteristics of clinicaly isolated and colonized linezolid(LZD)-resistant Enterococcus faecalis (E.faecalis) strains from a patient.Methods Ten E.faecalis strains (2 were isolated from urine specimens and 8 were from stool specimens) isolated from a patient with pulmonary infection were performed antimicrobial susceptibility testing, homology of E.faecalis was determined by pulsed-field gel electrophoresis (PFGE).Results Before and after patients received LZD therapy, 2 E.faecalis strains isolated form urine specimens were both resistant to LZD (MICs: 8 mg/mL, 16 mg/mL, respectively), among 8 strains from stool specimens (6 were isolated before therapy, and 2 were isolated after therapy), LZD susceptible, intermediate, and resistant strains were 4, 2, and 2 respectively(MICs: 0.25-12 mg/mL).10 strains of E.faecalis were homologous by PFGE typing.Conclusion In this case, the detection of E.faecalis from urinary tract and intestinal tract is homologous, which suggested that LZD-resistant Enterococcus may be colonized in vivo for a long time, and may be shift to cause bacterial infection.
5.Survey of rehabilitation therapy in clinical settings of Chongqing in 2015:some thoughts for local all-around development in rehabilitative profession
Xue DENG ; Xuelian WU ; Lin HAN ; Bin ZHENG ; Ying YIN ; Lehua YU
Chinese Journal of Medical Education Research 2016;15(12):1284-1290
Objective There is a huge demand for rehabilitative services in Chongqing whereas detailed information regarding local clinical services and rehabilitative professionals are unknown. Therefore, a cross-sectional survey was conducted to generate a profile of current rehabilitative status in Chongqing for reference of relevant local parties and peers in other provinces of China to create a larger picture of rehabil-itation in China. Methods The survey began in early January of 2015 and ended on October 20th of 2015. With two categories of information, including background of rehabilitation professionals and rehabilitative clinical services, the self-made questionnaire was sent via email to rehabilitation therapists working in local clinical settings which fulfilled the inclusion criteria. The reminder was sent once a month to those who did not yet replied. The software SPSS 20.0 was used to analyze the data descriptively. Results 7 out of 13 in-volved clinical settings responded with an overall response rate of 53.8%. 98 out of 122 copies were adopted based on filling requirement. The result revealed 68.4%(n=67) of the participants were junior practitioners, 57.1%(n=56) of them had less than 5 years experience and 45.9%(n=45) were holding a diploma with little research experience. As for clinical services, assessment of ROM (94.9%, n=93) and manual muscle testing (94.9%, n=93) were mostly used. The physical therapy service was more prevalent than occupational therapy services. The clinical field mostly involved was adult rehabilitation (77.6%, n=76). Bedside training in the acute phase (81.6%, n=80) and one-to-one treatment (92.9%, n=91) was the most commonly seen mode of service in the departments. Conclusion The local rehabilitative professionals were featured by their young age with comparatively low educational level, qualification and weak research skills. The clinical services they provided were yet quite basic and simple. Thus, a set of policies regarding education and profession in rehabilitation therapy should be facilitated instantly and it also calls for the establishment of local academic society for rehabilitation therapists, thus promoting an all-round development of rehabilitative profession.
6.Clinical evaluation of the (1, 3)-β-D-glucan assay as an aid to diagnosis of fungal infections in severe pneumonia patients
Wenxin ZENG ; Yuan HUANG ; Yu DENG ; Miaoyun WEN ; Yongli HAN ; Wenhong ZHONG ; Hongke ZENG
Chinese Journal of Emergency Medicine 2016;25(5):659-662
Objective To evaluate (1,3)-β-D-glucan (BG) assay as an aid for invasive fungal infection (IFI) diagnosis in severe pneumonia patients (diagnosis followed 2007 American Thoracic Society (ATS) and Infectious Disease Society of America (IDSA) severe pneumonia standard).Methods BG antigenemia was measured by BG Assay Box.IFIs was classified according to the blood fungal laboratory reports.Results 558 patients (185 females,373 males,mean age 64.7) were included.41 patients were proven to be fungal infected to be classified in exposure group.BG assay mean value in exposure group and unexposure group were (568.53 ±796.57) pg/mL,(51.4 ±63.27) pg/mL,respectively.Patients in the exposure group had significantly higher BG assay value than patients in the unexposure group (P <0.05).For the cutoff 100 pg/mL recommended by manufacturer,the sensitivity,specificity,positive predict value and negative predict value of the BG assay were 92.7%,92.5%,49.4% and 0.6%,respectively.Conclusion BG assay has positive clinical value in invasive fungal infection diagnosis in severe pneumonia patients.
7.Effects of IL-1β on expression of AQP4 and its role in attack of seizure
Zhen DENG ; Han YU ; Yuanshu ZHAO ; Guilin QI ; Meng MA ; Yalan LUO ; Xiaoqin ZHU ; Shuisheng LEI
The Journal of Practical Medicine 2016;32(5):698-701
Objective To observe the effects of interleukin-1β (IL-1β) and pentylenetetrazol (PTZ) induced acute epilepsy and the dynamic expression of aquaporin-4 (AQP4) in hippocampus. To explore the role of IL-1β in the pathogenesis of epilepsy by regulating AQP4. Methods All rats were randomly divided into control group, IL-1β group, PTZ group, IL-1ra + PTZ group and dexamethasone + PTZ group. Observe the behavior of the rats within 60 minutes after injection and record seizure score in each group. Then immunohistochemistry and RT-qPCR were used to detect the expression of AQP4 at at 6 , 12, 24 and 36 h. Results Almost of rats in IL-1β group and PTZ group showed severe degree seizure. The rats in control group and dexamethasone + PTZ group showed no obvious seizure. The seizure of rats were more remarkable serious in PTZ group than that in the IL-1ra + pentylenetetrazole group (P < 0.05). Immunohistochemistry and RT-qPCR Show: the expression of AQP4 in hippocampus in PTZ group increased gradually after 12 h (P < 0.05), then reached in the peak after 24 h (P < 0.001). The expression of AQP4 in IL-1ra + PTZ group was lower compare with PTZ group in each time (P < 0.05). Although the expression of AQP4 in dexamethasone + PTZ group higher than the control group, it was not significantly different (P < 0.05). Conclusion The proinflammatory cytokine IL-1β break the balance of water in brain and increasing the concentration of extracellular excitatory amino acids or ions by upregulate the expression of AQP4 in order to promote the excitatory of neurons.
8.Construction of a recombinant adenovirus vector expressing shRNA targeting interleukin-1beta gene in rats
Xiaolong ZHAO ; Jianping CHEN ; Yu ZHANG ; Hang LI ; Yanfang LIU ; Wenyan GAO ; Lei HAN ; Yanan DENG
Chinese Journal of Tissue Engineering Research 2015;(18):2923-2927
BACKGROUND:Specific down-regulation of interleukin 1 beta (IL-1β) may al eviate the pain behaviors effectively after peripheral nervous injury. Compared with smal interference RNA (siRNA), short hairpin RNA (shRNA) could inhibit the expression of target gene more stably and efficiently. However, simple shRNA could not enter target cel s to down-regulate target gene efficiently. Adenovirus vectors have wide host range, high infection efficiency and stable expression in host cel s.
OBJECTIVE:To construct recombinant adenovirus vector expressing shRNA targeting IL-1βgene and detect its effect on the expression of target gene.
METHODS:Three siRNAs were designed on the basis of the nucleotide sequence of IL-1βobtained from NCBI and then three shRNAs (shRNA1, shRNA2 and shRNA3) were synthesized. The annealed shRNA product and adenovirus vector pHBAd/U6/GFP digested by BamH I and EcoR I were connected to construct the recombinant adenovirus vector shuttle plasmid expressing shRNA targeting IL-1β. After sequencing, HEK 293 cel s were co-transfected by the shuttle plasmid and skeleton vector, and three recombinant adenovirus vector expressing shRNA targeting IL-1β(rAd/shRNA1, rAd/shRNA2 and rAd/shRNA3) were packaged and amplified. Rats H9C2 cel s were infected by recombinant adenovirus vector expressing shRNA targeting IL-1βand fluorescence microscope was used to observe the infection efficiency. The effect of recombinant adenovirus vector expressing shRNA targeting IL-1βon the expression of target gene was detected by western blot assay.
RESULTS AND CONCLUSION:The sequencing results showed that the sequences of three shRNAs adenovirus vector shuttle plasmid were consistent with the sequences of three designed shRNAs. rAd/shRNA1, rAd/shRNA2 and rAd/shRNA3 were constructed successful y. rAd/shRNA1, rAd/shRNA2 and rAd/shRNA3 could down-regulate the expression of IL-1βin rat H9C2 cel s and the down-regulation effect of rAd/shRNA2 was the most significant.
9.Transcatheter radiofrequency ablation of atrial tachycardia originating from the side of the bundle of His
Zhihua YU ; Hongwei HAN ; Guanghui CHENG ; Zhen LI ; Jinglin ZHANG ; Chenggang DENG ; Xi SU
Journal of Interventional Radiology 2015;(9):759-762
Objective To investigate the electrophysiological characteristics of atrial tachycardia (AT) originating from the side of the bundle of His, and to analyze the efficacy and safety of transcatheter radiofrequency ablation via the right side of the bundle of His and left noncoronary sinus of aorta for the treatment of AT. Methods A total of 12 patients with AT originating from the side the bundle of His, which was confirmed by conventional electrophysiological study and atrial activation patterns, were included in this study. The patient’s age ranged from 12 to 64 years old with a mean of (47.4±14.6) years. The mapping was carried out at the right side of the bundle of His and radiofrequency ablation was performed. When the ablation procedure failed, or the junction zone rhythm or atrioventricular block occurred, ablation via the left noncoronary sinus of aorta was employed. Results Atrial stimulation could repeatedly induce and terminate AT in all the 12 patients, the average cycle length was (327±76) ms. TA was terminated within 10 seconds after the start of ablation in 10 patients during their AT attacks. Ablation was unsuccessful in 2 patients. Ablation via the right side of His bundle was successfully accomplished in 2 patients, and the ablation via the left noncoronary sinus of aorta was successfully carried out in 8 patients. The average follow-up time was 1-6 years, and no recurrence of AT was observed. Conclusion Atrial tachycardia originating from the side of the bundle of His has certain electrophysiological characteristics, and transcatheter radiofrequency ablation is safe and effective for its treatment. Ablation via the left noncoronary sinus of aorta should be considered as a strategy of priority.
10.Changes in phosphorylation of NMDA receptors in spinal dorsal horns in a rat model of neuropathic pain induced by chronic compression of dorsal root ganglion
Jianping Chen ; Chongfang Han ; Yu Zhang ; Yanfang Liu ; Xiaolong Zhao ; Ya'nan Deng
Chinese Journal of Anesthesiology 2014;34(5):566-569
Objective To investigate the changes in the phosphorylation of N-methyl-D-aspartate (NMDA) receptors in spinal dorsal horns in a rat model of neuropathic pain induced by chronic compression of dorsal root ganglion (CCD).Methods Ninety-six male Sprague-Dawley rats,weighing 200-250 g,were randomly divided into 3 groups (n =32 each) using a random number table:control group (group C),sham operation group (group S) and group CCD.In CCD group,a small stainless steel needle (4 mm in length and 0.8 mm in diameter) was inserted into the L4,5 intervertebral foramen in pentobarbital sodium-anesthetized rats,developing intervertebral foramen stenosis and hence producing a chronic steady compression of the dorsal root ganglion.In group S,the intervertebral foramen was only exposed without inserting the needle.The paw withdrawal latency (PWL) was measured at 1 day before operation and 1,2 and 4 weeks after operation.The rats were then sacrificed after measurement of PWL and the lumbar enlargement segments of the spinal cord were removed for determination of the expression of interleukin-1β (IL-1β) and phosphorylation of NMDA receptor NR1 subunits at serine 896 (pNR1S896) on the injured side by immunohistochemistry and Western blot.Results Compared with group C,the PWL was significantly shortened at each time point after operation,and the expression of IL-1β and pNR1S896 was up-regulated in group CCD,and the PWL was shortened at 1 week after operation,the expression of IL-1β and pNR1S896 was up-regulated,and no significant change was found in PWL and expression of IL-1β and pNR1S896 at the other time points in group S.Compared with group S,the PWL was significantly shortened at each time point after operation,and the expression of IL-1β and pNR1S896 was up-regulated in group CCD,Compared with the baseline value at 1 day before operation,the PWL was slowly shortened starting from 1 week after operation,and decreased to the lowest level at 2 weeks after operation and maintained at this level for 2 weeks,the expression of IL-1β and pNR1S896 was slowly up-regulated starting from 1 week after operation,and increased to the highest level at 2 weeks after operation and maintained at this level for 2 weeks in group CCD,and the PWL was shortened at 1 week after operation,and the expression of IL-1β and pNR1 S896 was up-regulated in group S.Conclusion The development and maintenance of neuropathic pain induced by CCD are related to phosphorylation of NMDA receptors in spinal dorsal horns of rats.