1.Establishment of a Stable Cell Line Expressing"Toxic" Transient Receptor Potential A1 Channel
Shaopeng WANG ; Lianghui MA ; Jixian LI ; Zhigang PENG ; Jing CAO
Progress in Biochemistry and Biophysics 2008;35(12):1378-1386
Transient receptor potential A1 (TRPAl) is a cold sensitive cation channel, which could also be activated by various pungent compounds. As a transduction channel in a number of sensory modalities, TRPAl expressing in heterogonous systems serves to provide great convenience in pharmacological analysis and functional investigation. Due to cellular toxicity, establishment of stable TRPAl cell line has always been challenging. Nevertheless, the first stable human embryonic kidney (HEK-293) cell line with un-controlled expression of TRPAl was successfully established. It was also confirmed that this stable cell line retained TRPAl expression for more than 25 passages in culture. The functional analysis of the cell response verified the stability and specificity of this novel recombinant TRPAl cell line. Altogether, the data indicated this TRPAl-HEK cell line would be a useful tool for functional analysis of TRPAl and for the development of high throughput screening (HTS) compatible assay in the effort to identify TRPAl modulators.
2.Non-enhanced CT axis rotating movie imaging in percutaneous nephrolithotomy for complex renal calculi
Rongpei WU ; Zhenpeng PENG ; Xiaofei LI ; Shaopeng QIU ; Chaogui YAN ; Lingwu CHEN
Chinese Journal of Urology 2010;31(3):165-168
Objective To discuss the clinical application and significance of non-enhanced computed tomography axis rotating movie imaging technique in PCNL for complex renal calculi. Methods Thirty-one cases unilateral and 2 cases bilateral multiple and staghorn renal calculi with mild or mediurn hydronephrosis patients were performed bilateral kidneys non-enhanced CT scanning,three dimensional reconstruction and the axis rotating movie composition were carried on by computer software,PCNL accesses were designed and the residual stone were predicted referred to the access-calyces angle measured in axis rotating movie image,PCNL were performed after while.Comparing between preoperation accesses design and residual stone prediction with in-operation practice were carried out.Results The first PCNL access was constructed via posterior middle upper minor calyces in 22 renal units and via posterior middle lower minor calyces in 13 renal units,which was consistent with pre-operation design according to CT axis rotating movie image.The second PCNL accesses were constructed via lower calyx posterior upper minor calyces in 9 renal units and via lower calyx posterior lower minor calyces in 5 renal units,nephrolithotomy were performed in the same operation,clinical stone clearance rate was 80%(28/35),other 7 cases with residual stone were consistent with pre-operation prediction,No blood transfusion was necessary and no severe complication happened in all 33 cases.Conclusions Non-enhanced CT axis rotating movie imaging provided the detail three dimensional shape and spatial structure of complex renal calculi intuitively) that was benefit for designing appropriate PCNL accesses for complex renal calculi patients, guiding for searching stone fragments in operation, predicting residual stone, and ensuring operation safety.
3.Establishment of a cell based high throughput screening model for transient receptor potential V3 modulators.
Shaopeng WANG ; Jixian LI ; Zhigang PENG ; Jing CAO
Chinese Journal of Biotechnology 2008;24(11):1895-1901
We established a cell based high throughput screening model by calcium assay on fluorometric imaging plate reader for finding modulators of TRPV3. The TRPV3 expression vector was transfected into HEK-293 and stable cell line expressing TRPV3 was selected with antibiotics. Upon TRPV3 specific modulators stimulated, pharmacological characteristics of TRPV3 over expression cell line were detected by calcium assay on fluorometric imaging plate reader. Assay conditions were optimized and stability of the model was observed. The reliability and accuracy of application to 96 and 384 well format high throughput screening were also evaluated. A stable HEK-293 cell line highly expressing TRPV3 was established. TRPV3 specific modulators could modulate calcium signal through TRPV3 in dose dependent manner. Optimized screening condition was established by assay development. This model is stable and sensitive, and meets the requirement of high throughput screening by Z'factor validation and Spiking test. This cell model can be applied to screening TRPV3 modulators by calcium assay.
Calcium
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metabolism
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Cell Line
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Humans
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Ion Transport
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Kidney
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cytology
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Membrane Transport Modulators
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metabolism
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Models, Biological
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TRPV Cation Channels
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biosynthesis
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genetics
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Transfection
4.Clinical observation and effects by the methods of Xingshen-Kaiqiao to treat diffuse axonal injury combined with western medicine normal treatment
International Journal of Traditional Chinese Medicine 2018;40(7):592-595
Objective In order to observe the clinical curative effect of magnesium sulfate combined with Xingshen-Kaiqiao acupuncture for the diffuse axonal injury (DAI). Methods A total of 60 patients with DAI were divided into the Xingshen-Kaiqiao group and control group according to the random number table method (30 in each). The control group was given traditional Chinese medicine wash-out on the basis of conventional therapy, saline 100 ml+25% magnesium sulfate 8 ml and slow intravenous infusion, then 5% glucose 500 ml+25% magnesium sulfate 8 ml and slow intravenous drip; during the duration of the drug, the concentration of blood magnesium was strictly monitored and the dosage was adjusted as appropriate. And the Xingshen-Kaiqiao group was given acupuncture of Xingshen-Kaiqiao on the basic treatment of the control group. All treatments lasted 4 weeks. After treatment, the expression levels of IL-1β and TNF-α, GSC score, recovery of consciousness and GOS were evaluated and compared before and after treatment. Results After treatment, the IL-1β (62.38 ± 16.92 pg/ml vs. 88.37 ± 14.71 pg/ml, t=8.554), the TNF-α (3.59 ± 1.05 pg/ml vs. 4.06 ± 1.62 pg/ml, t=5.081) of Xingshen-Kaiqiao group were significantly lower than those of the control group (P<0.05). The GSC scores after 1st week (8.26 ± 0.76 vs. 6.83 ± 0.82, t=-5.036), 2nd week (10.61 ± 0.82 vs. 8.91 ± 0.35, t=-4.387), 3rd week (12.52 ± 1.07 vs. 10.95 ± 0.67, t=-5.212), 4th week (13.26 ± 1.08 vs. 11.58 ± 1.86, t=-5.031) of Xingshen-Kaiqiao group were significantly higher than those of the control group ( P<0.05). After treatment, the Xingshen-Kaiqiao group awakening rate was 70.0% (21/30) and the control group was 43.3% (13/30), and the difference in awakening rate was statistically significant (χ2=4.344, P=0.037). The awakening time (7.6 ± 3.1 d vs. 11.5 ± 4.9 d, t=6.586) of Xingshen-Kaiqiao group was significantly lower than the control group (P<0.05); the percentage of GOS was significantly higher than the control group (Z=-2.093, P=0.036). Conclusions Magnesium sulfate combined with acupuncture of Xingshen-Kaiqiao can reduce the awakening time, the death rate and showe more satisfactory clinical efficacy for the patients with DAI.
5.Effect of sevoflurane anesthesia on expression of hippocampal α4nAchR in rats
Siqin LUO ; Kaiyun FANG ; Xiangping JIANG ; Shaopeng GANG ; Yi MA ; Lin PENG ; Xiang HE
Chinese Journal of Anesthesiology 2018;38(2):163-167
Objective To evaluate the effect of sevoflurane anesthesia on the expression of hippocampal α4 subunit-containing nicotinic acetylcholine receptor (α4nAChR) in rats.Methods One hundred and forty-four Sprague-Dawley rats of both sexes,aged 3-4 months,weighing 220-270 g,were divided into 4 groups (n =36 each) using a random number table:control group (group C),sevoflurane anesthesia for 1 h group (group S1),sevoflurane anesthesia for 3 h group (group S2) and sevoflurane anesthesia for 5 h group (group S3).Group C inhaled air,and S1,S2 and S3 groups inhaled 3% sevoflurane for 1,3 and 5 h,respectively.Twelve rats in each group were selected at 1 and 7 days after emergence from anesthesia to undergo spatial probe test.Rats were then sacrificed immediately after anesthesia and at 1 and 7 days after emergence from anesthesia,and hippocampi were removed for determination of the expression of α4nAchR protein and mRNA in hippocampal neurons (by Western blot or real-time polymerase chain reaction).Results Compared with group C,the duration of staying at the target quadrant was significantly shortened,and the ratio of duration of staying at the original platform quadrant to the total duration and ratio of swimming distance in the original platform quadrant to the total distance were decreased on 1 and 7 days after emergence from anesthesia,the expression of α4nAchR protein and mRNA was down-regulated,and the number of positive cells was reduced in S1,S2 and S3 groups (P<0.05).Compared with S1 and S2 groups,the duration of staying at the target quadrant was significantly shortened,the ratio of duration of staying at the original platform quadrant to the total duration and ratio of swimming distance in the original platform quadrant to the total distance were decreased on 1 day after emergence from anesthesia in group S3 (P<0.05).There was no significant difference in the expression of α4nAchR protein and mRNA or number of positive cells at each time point between group S1,group S2 and group S3 (P>0.05).Conclusion The mechanism by which sevoflurane anesthesia induces cognitive dysfunction may be partially related to down-regulating the expression of hippocampal α4nAchR in rats.
6.Relations of balloon pressures with complications and recurrence in treatment of trigeminal neuralgia by percutaneous balloon compression
Shaopeng PENG ; Jianxiong LIU ; Fei WANG ; Hongxing ZHANG ; Jian ZHANG
Chinese Journal of Neuromedicine 2022;21(2):152-156
Objective:To investigate the changing characteristics and stages of balloon pressures in treatment of trigeminal neuralgia by percutaneous balloon compression (PBC), and explore the relations of balloon pressures at different stages with postoperative facial numbness and pain recurrence.Methods:A prospective analysis was performed. Thirty-two patients with primary trigeminal neuralgia, admitted to our hospital from May 2019 to May 2020, were chosen. During surgery, a manometer was connected with computer manometry software to continuously monitor the balloon pressure changes; pear-shaped stage and maintenance stage were named during the balloon pressure changes. The facial numbness occurrence and pain recurrence were assessed by Barrow Institute of Neurology (BNI) facial numbness and pain intensity scales. Robust evaluation method referred to American Institute for Clinical and Laboratory Standardization (CLSI) C28-A2 document was used to determine the reference ranges of balloon pressures at the pear-shaped stage, maintenance stage and whole-process stage (pear-shaped stage+maintenance stage). Spearman rank correlation test was used to analyze the correlations of balloon pressures at the pear-shaped stage, maintenance stage and whole-process stage with facial numbness 24 h and 12 months after surgery, and pain recurrence 6 and 12 months after surgery.Results:Among the 32 patients, pain relief was found in 31 patients and lateral numbness was found in 30 patients 24 h after surgery. Pain recurred 12 months after surgery in 2 patients. The reference ranges of pressure at pear-shaped stage, maintenance stage and whole-process stage were (157.2-128.5) kPa, (136.10-117.9) kPa and (141.9-119.9) kPa, respectively. The pressures at pear-shaped stage, maintenance stage and whole process stage were positively correlated with facial numbness 24 h and 12 months after surgery ( P<0.05); there were negative correlations between pressures at pear-shaped stage and whole-process stage and pain recurrence 6 months after surgery ( P<0.05); pressures at pear-shaped stage, maintenance stage and whole-process stage were negatively correlated with pain recurrence 12 months after surgery ( P<0.05). Conclusion:The lower the balloon pressure during PBC, the more likely it would have pain recurrence after surgery; and the higher the pressure, the more likely it would have facial numbness after surgery; the efficacy of PBC can be maximized when the balloon pressures maintains at adequate ranges ([157.2-128.5] kPa at the pear-shaped stage, [136.1-117.9] kPa at the maintenance stage and [141.9-119.9] kPa at the whole-process stage).
7.Perioperative factors affecting the length of hospital stay after cesarean section of pregnant women with heart disease
Jing PENG ; Kaiyun FANG ; Zhenghua WANG ; Shaopeng GANG ; Chenguang QIN ; Fujuan HE ; Naiying SHANG ; Hongbi SONG ; Yan ZHU
Chinese Journal of Obstetrics and Gynecology 2021;56(6):408-417
Objective:To explore the related factors influencing the length of hospital stay(LOS) of pregnant women with heart disease (PWHD) after cesarean section.Methods:A total of 306 patients with PWHD who underwent cesarean section from January 2012 to March 2019 were collected. Among them, 203 patients had not undergone heart surgery (uncorrected group) and 103 patients who had undergone heart surgery (corrected group) during the same period. Demographic, perioperative and postoperative data were recorded. Predictors associated with postoperative LOS were determined using univariate and multivariate linear regression analysis models.Results:(1) The median LOS after cesarean section in the uncorrected group was 6 days (5-8 days). The results of univariate linear regression analysis showed that 38 parameters had significant impact on LOS ( P<0.05). The results of multivariate linear regression analysis showed that 5 parameters were independent risk factors for prolonged LOS in the uncorrected group; among them, the median LOS in uncorrected group with hypertensive disorders of pregnancy was 3 days longer than that in patients with PWHD alone [7 days (5-8 days) vs 4 days (4-5 days), β=0.195, P=0.001]; the median LOS in uncorrected group with high serum creatinine was 3 days longer than normal patients [7 days (5-13 days) vs 4 days (4-5 days), β=0.145, P=0.015]; the LOS of patients who chose general anesthesia was 2 days longer than that of patients who chose spinal anesthesia [6 days (4-8 days) vs 4 days (4-5 days), β=0.154, P=0.007]; the LOS of patients with postoperative pulmonary infection was 4 days longer than that of patients without pulmonary infection [8 days (5-15 days) vs 4 days (4-5 days), β=0.269, P<0.01]; the LOS of patients who admitted to ICU after surgery was 2 days longer than that not admitted patients [6 days (5-8 days) vs 4 days (4-5 days), β=0.268, P<0.01]. (2) The median LOS after cesarean section in corrected group was 4 days (4-5 days). The results of univariate linear regression analysis showed that 8 parameters had significant impact on the LOS (all P<0.05). The results of multivariate linear regression analysis showed that 2 parameters, which were American Society of Anesthesiologists (ASA) grade ( β=0.198, P=0.028) and intraoperative blood loss ( β=0.285, P=0.003), were the independent risk factors for prolonged LOS in corrected group. Conclusion:Preoperative with hypertensive disorders of pregnancy, preoperative creatinine increase, intraoperative general anesthesia, postoperative pulmonary infection, and postoperative admission to ICU are independent predictors of prolonged LOS in uncorrected patients with PWHD; ASA classification and intraoperative bleeding are independent predictor of prolonged postoperative LOS in patients with corrected PWHD.
8. Incidence and risk factors of postoperative acute renal injury in patients underwent hip and knee replacement
Yi MA ; Kaiyun FANG ; Shaopeng GANG ; Jing PENG ; Ling JIANG ; Li SUN ; Yan ZHU
Chinese Journal of Orthopaedics 2019;39(19):1192-1198
Objective:
To investigate the incidence and risk factors of postoperative acute renal injury (AKI) in patients after primary unilateral hip and knee replacement.
Methods:
Between January 2016 and July 2018, A total of 1 490 patients underwent hip and knee arthroplasty were enrolled. There were 507 male and 983 female with a median age of 66 years (range: 18 to 95 years). There were 201 patients underwent femoral head replacement, 897 total hip arthroplasty and 392 total knee arthroplasty, respectively. The creatinine value was 68.79 μmol/L preoperatively. Demographic, perioperative and postoperative data were recorded. The development of AKI was defined according to Kidney Disease Improving Global Outcomes (KDIGO) criteria. The demographic information, comorbidities, preoperative medication, type of surgery, type of anesthesia, intraoperative anesthetics, operation time, intravenous fluid type and dose, amount of bleeding and postoperative creatinine values were recorded. Predictors associated with AKI and postoperative creatinine was determined by multiple regressions.
Results:
There were 80 cases developed AKI with the incidence of AKI 5.4%. Among the AKI patients, there were 69 cases (4.6%) in AKI stage 1, 7 cases (0.5%) in AKI stage 2, and 4 cases (0.3%) in AKI stage 3. The creatinine value was 72.03 μmol/L after surgery. The length of postoperative hospital stay was 9 days. There were 5 patients died in hospital. The independent risk factors of AKI after hip and knee arthroplasty included old age, a higher American Society of Anesthesiologists (ASA) grade and preoperative diabetes (