1.Soluble expression of tandem hybrid ubiquitin-binding domains (ThUBD) in prokaryotic cytoplasm of Escherichia coli BL21(DE3)
Weijia LUO ; Chen DENG ; Yanchang LI ; Yuan GAO ; Ping XU
Military Medical Sciences 2016;40(10):795-800,818
Objective We increase the soluble expression of artificial tandem hybrid ubiquitin binding domains ( ThUBD) in prokaryotic cytoplasm of Escherichia coli BL21 ( DE3 ) , which offer an effective and special profiling for ubiquitin conjugates( UbC) .Methods Codon optimization of the ThUBD was performed, followed by analysis of codon relative adaptiveness based on relative frequency of synonymous codon ( RFSC) of E.coli.Further induced expression and yeast ubiquitin conjugate enrichment quantified the soluble ThUBD-S and tested the ability to bind UbC.Results The statistical result showed that the percentage of codon of the highest usage frequency was increased from 48%to 75%, and codon adaptation index( CAI) was increased from 0.63 to 0.88 after codon optimization, which might suggest a higher expression of the ThUBD in E.coli BL21 (DE3).The subsequent SDS-PAGE indicated that the soluble target protein was increased four times, which accounted for 13.06%of total cell lysis.Further ubiquitinated proteome of yeast demonstrated that the ability to bind and enrich UbC of optimized ThUBD-S did not change compared with original ThUBD.Conclusion The expression of ThUBD-S can quadruple after codon optimization.At the same time, codon optimization does not impact its soluble expression and the ability to bind UbC.
2.Expression of serum IL-6 and IL-10 in sepsis patients and their impacts on immune function
Rui FAN ; Jian SHEN ; Weijia DOU ; Ping CAO ; Jia YUAN ; Youwen YE
International Journal of Laboratory Medicine 2015;(22):3250-3252
Objective To study the expression of serum interleukin (IL )-6 and IL-10 in sepsis patients ,and their correlations with immune function .Methods A total of 56 patients with sepsis were divided into three groups ,including 18 patients in sepsis group ,21 patients in severe sepsis group ,17 patients in septic shock group .All the patients were also divided into survivor and death group according to whether they survived within 2 weeks .Other 30 healthy persons were selected in the control group .Serum IL-6 and IL-10 ,CD4+ /CD8+ were detected and compared .Results Serum IL-6 and IL-10 levels in the three sepsis groups were significantly higher than those in control group ,and with the severity aggravating ,these indicators increased .The level of serum IL-6 in sepsis patients was significantly reduced at the 3rd day ,while there was no difference on the serum IL-10 in severe sepsis group and septic shock group compared with that at admission(P>0 .05) .Compared with survivor group ,serum IL-6 and IL-10 levels on both admission and the 3rd in death groups increased significantly (P<0 .05) .CD4+ ,CD8+ ,CD4+ /CD8+ were negative related with serum IL-6 and IL-10 in both severe sepsis and septic shock group (P<0 .05) .Conclusion The expression of IL-6 and IL-10 in patients with sepsis are over-expressive ,and closely related with disease activity and immune function .
3.Effects of non-invasive limb ischemic preconditioning on endothelial function after myocardial ischemia-reperfusion injury
Weijia CHEN ; Dongming LIU ; Zhaoyan QIANG ; Ke WEN ; Hengjie YUAN ; Yi KANG ; Jianshi LOU ; Yanna WU
Chinese Pharmacological Bulletin 2014;(12):1692-1697
Aims To investigate the protective effects of noninvasive limb ischemic preconditioning (LIPC) on myocardial ischemia-reperfusion (I /R)injury,and to explore the mechanism.Methods Healthy male Wistar rats were divided randomly into I /R,I /R +LIPC,I /R +5-Hydroxydecanoate (5-HD)and I /R +LIPC +5-HD groups.The I /R +LIPC and I /R +LIPC-5-HD groups of rats were subjected to three cy-cles of LIPC induction per day with 5 min of reperfu-sion after occlusion for 5 min at the left hind limb for 3 days.All rats were subjected to myocardial I /R injury on the fourth day.The I /R +5-HD and I /R +LIPC-5-HD groups of rats were given the inhibitor of ATP-sen-sitive potassium channel 5-HD before and during myo-cardial I /R injury. Results Compared with I /R group,LIPC reduced myocardial infarct size (P <0.05),lowered cardiocyte apoptosis index and Fas, FasL positive cell number (P <0.01 ),increased the reduced nitric oxide (NO)/endothelin (ET)-1 ratio (P <0.05)in serum in I /R +LIPC group.5-HD a-bolished the protective effects induced by LIPC in I /R+LIPC-5-HD group.Compared with normal myocardi-al tissue,expression of mir-30a-3p was increased in I /R group (P <0.01 )and was decreased in LIPC group (P <0.01 ).Conclusion LIPC alleviates myocardial I /R injury and improves endothelial function. The mechanism may be related with the opening of ATP-sensitive potassium channel,regulating the balance be-tween NO and ET-1 and decreasing the expression of myocardial mir-30a-3p.
4.Effects of different biliary drainage methods on liver regeneration of obstructive jaundice rats after partial hepatectomy
Shengguang YUAN ; Kewei LIANG ; Jie LIU ; Weijia LIAO ; Liling QIN ; Songqing HE
Chinese Journal of Digestive Surgery 2013;12(12):956-962
Objective To investigate the effects of internal and external biliary drainage on liver regeneration of the obstructive jaundice rats after partial hepatectomy.Methods The rat models of obstructive jaundice with 70% liver resection were successfully constructed.All the 120 rats were randomly divided into the control group:rats received middle and left hepatic lobectomy; internal drainage group:a drainage tube was placed between the dilated bile duct and the duodenum; external drainage group:a drainage tube was placed in the dilated bile duct.There were 40 rats in each group.Rats in the internal and external drainage groups received middle and left hepatic lobectomy at postoperative day 7.The blood and hepatic tissues were collected at postoperative day 0,1,2,4,12,24,48,72 hours after operation,and the rate of liver regeneration and mitotic index were determined.The expression of proliferating cell nuclear antigen (PCNA) and signal transducer and activator of transcription 3 (STAT3) in the hepatic tissues were detected by immunohistochemistry,and the levels of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) were detected by ELISA,and the mRNA expressions of TNF-α and IL-6 were detected by RT-PCR.All data were analyzed using the one way analysis of variance or SNK test.Results Within 72 hours after partial hepatectomy,the rate of liver regeneration of the internal drainage group was 94.86%± 12.72%,which was significant higher than 62.39%±8.01% of the external drainage group and 45.77% ± 5.41% of the control group (F =33.62,P < 0.05).The mitotoic index and PCNA levels of the 3 groups had obvious increase at postoperative hour 12,and the mitotoic index and PCNA levels of the external drainage group reached peak at postoperative hour 24,which were 24.47% ± 4.01% and 88.1% ± 9.2%,respectively,the mitotoic index and PCNA levels of the control group and the external drainage group reached peak at postoperative hour 48,which were 15.80% ± 1.08%,58.3% ± 5.8% and 18.40% ± 1.12%,70.2% ± 6.9%,respectively.The mitotoic index and PCNA levels of the internal drainage group were significantly higher than those of the control group and the external drainage group (P < 0.05).The expression of STAT3 expression of the internal drainage group reached peak at postoperative hour 4,which was 42.6% ± 3.6% ;the expression of STAT3 expression of the control group and the external drainage group reached peak at postoperative hour 12,which were 22.9% ± 2.0% and 29.2%± 3.7%.The peak level of STAT3 of the internal drainage group was significantly higher than those of the control group and the external drainage group (P <0.05).The levels of TNF-α and IL-6 of the internal drainage group reached peak at postoperative hour 12,which were (227 ±23)U/L and (256 ± 32)U/L; the levels of TNF-α and IL-6 of the control group and the external drainage group reached peak at postoperative hour 24,which were (309 ± 41) U/L and (388 ± 40) U/L,(287 ± 30)U/L and (346± 33)U/L,respectively.The levels of TNF-α and IL-6 of the internal drainage group at postoperative hour 0,1,2,4,12,24,48,72 were significantly lower than those of the control group and the external drainage group (P < 0.05).The expressions of TNF-α mRNA of the control group,internal drainage group and external drainage group reached peak at postoperative hour 4,which were 0.92 ±0.14,0.39 ±0.05,0.80 ±0.15,respectively.The expressions of IL-6 mRNA reached peak at postoperative hour 12,which were 0.79 ± 0.07,0.38 ± 0.06,0.63 ±0.10,respectively.The expressions of TNF-α mRNA and IL-6 mRNA of the internal drainage group at postoperative hour 0,1,2,4,12,24,48,72 were significantly lower than those of the control group and the external drainage group (P < 0.05).Conclusions Both internal and external drainage can improve liver regeneration of obstructive jaundice rats following partial hepatectomy,while the effect of internal drainage is superior.Internal biliary drainage has influence on the expression of STAT3 by decreasing the levels of TNF-α and IL-6,and help to improve liver regeneration of obstructive jaundice rats following partial hepatectomy.
5.Antidiabetic drugs of plant origin used in China: compositions, pharmacology, and hypoglycemic mechanisms.
Wei JIA ; Wen-yuan GAO ; Pei-gen XIAO
China Journal of Chinese Materia Medica 2003;28(2):108-113
The paper reviewed compositions and pharmacological effects of eight antidiabetic herbal drugs that have been approved by health regulatory agency for commercial use in China. Investigators attributed the hypoglycemic effect of these products to their ability to restore the functions of pancreatic tissues and cause an increase in insulin output, to inhibit the intestinal absorption of glucose, or to the facilitation of metabolites in insulin-dependent processes. Treatment with herbal drugs has an effect on protecting beta cells and smoothing out fluctuations in glucose levels. The use of these naturally derived agents in conjunction with conventional drug treatments such as an chemical agent or insulin permits the use of lower doses of the drug and/or decreased frequency of administration which decreases the side effects most commonly observed.
Animals
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Astragalus Plant
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chemistry
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Blood Glucose
;
metabolism
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Diabetes Mellitus
;
drug therapy
;
metabolism
;
Drug Combinations
;
Drugs, Chinese Herbal
;
chemistry
;
isolation & purification
;
pharmacology
;
therapeutic use
;
Humans
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Hypoglycemic Agents
;
administration & dosage
;
pharmacology
;
therapeutic use
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Insulin Resistance
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Panax
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chemistry
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Phytotherapy
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Plants, Medicinal
;
chemistry
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Rehmannia
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chemistry
6.Effect of lumbar plexus-sacral plexus block combined with dexmedetomidine on rehabilitation of elderly patients undergoing PFNA
Junyu ZHOU ; Han ZHANG ; Weijia YUAN ; Xia YUAN ; Wei DAI ; Kaihua HE
Chongqing Medicine 2018;47(12):1616-1619,1624
Objective To evaluate the effectiveness of lumbosacral plexus block combined with the use of dexmedetomidine in elderly patients undergoing proximal femoral nail antirotation (PFNA).Methods A total of 60 patients received elective PFNA were divided into tracheal intubation combined with inhalation anesthesia group (group G) and ultrasound and nerve stimulator-guided lumbosacral plexus block following with dexmedetomidine infusion group (group N).Then we observed HR,SBP,DBP for both groups at the time entering the theater (T0),immediately after tracheal intubation or after dexmedetomidine infusion (T1),skin incision moment (T2) and 30 minutes after skin incision (T3).Visual analogue scale (VAS) scores were assessed for both groups at the time point of 2,6,12,24 and 48 hours after surgery.The number of use of patient controlled intravenous analgesia (PCIA),assessment of consciousness status 1-3 days after surgery,adverse reactions were recorded for both groups as well.The following post-surgery data were recorded:the time of first feeding,first urination and first ambulation,the length of hospitalization,the expense of hospital stay.Results HR,SBP,DBP of the group G changed more significantly at T1,T2,T3 than those of T0 (P<0.05).The VAS scores and the number of use of PCIA of group N were lower than those of group G at all time points after operation (P<0.05).The group N had lower CAM-CR scores and less adverse reactions of nausea and vomiting and dizziness than those of group G on days 1 to 3 after surgery (P<0.01).Compare to group G,the group N were early in terms of post-operation first feeding,first urination and first ambulation (P<0.01).The length of hospitalization was shorter and the cost of the hospital stay was lower in the group N than the group G (P<0.01).Conclusion Ultrasound and nerve stimulator-guided lumbosacral plexus block combined with low dose of dexmedetomidine could meet the needs of elderly patients undergoing PFNA.
7.The study of the high sensitivity to noise trauma in the inner ear mimetic aging model.
Li XU ; Yujuan HU ; Weijia KONG ; Hong SHI ; Wei PENG ; Yuan HU
Journal of Clinical Otorhinolaryngology Head and Neck Surgery 2010;24(19):895-900
OBJECTIVE:
Use the inner ear mimetic aging model which has been established by our research institute, investigate its sensitivity to noise trauma and the possible role of mitochondrial DNA deletions.
METHOD:
Thirty-two Wistar rats of 2 months old were randomly divided into four groups. In group A, D-galactose was subcutaneously injected at dose of 150 mg/kg weigh per day for 8 weeks, after that these rats were exposed to 110 dB SPL noise 4 hours each day, for 2 days. Group B were given normal saline (NS) injected at dose of 150 mg/kg weigh per day for 8 weeks,also given noise exposure as that of group A. Group C were give D-galactose without noise exposure. Group D were given normal saline (NS) without noise exposure. The thresholds of auditory brainstem response (ABR) were measured 2 weeks after stopping of noise exposure. And T-SOD and MDA of the inner membranous labyrinthine tissue were measured. Nest polymerase chain reaction (Nest PCR) were used to identify the mtDNA common deletion (CD), and PCR products were sequenced in the meantime.
RESULT:
The elevation of the mean ABR thresholds in group A was higher than that in group B, and the difference had statistic significance (P < 0.01). The reduction of T-SOD in group A was obvious, while the level of MDA was greatly increased. The difference in the levels of T-SOD and MDA between group A and group B had statistic significance (P < 0.01). The detection rate of mtDNA 4834 deletion were as follows: group A 87.5% (7/8); group B 12.5% (1/8); group C 75.0% (1/8); group D 0(0/8).
CONCLUSION
The rat in the inner ear mimetic aging model are hypersensitive to noise exposure, and mtDNA4834 deletions in the inner ear may play an important role in it.
Aging
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Animals
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DNA, Mitochondrial
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genetics
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Disease Models, Animal
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Disease Susceptibility
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Ear, Inner
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pathology
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Evoked Potentials, Auditory, Brain Stem
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Female
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Hearing Loss, Noise-Induced
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etiology
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genetics
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pathology
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Malondialdehyde
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analysis
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Noise
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Rats
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Rats, Wistar
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Sequence Deletion
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Superoxide Dismutase
;
analysis
8.Efficacy of velopharyngoplasty for treating obstructive sleep apnea hypopnea syndrome.
Wenzhong BAI ; Jianjun SUN ; Weijia KONG ; Wei YUAN ; Yu HU ; Rong ZHANG ; Yumei SUN
Journal of Clinical Otorhinolaryngology Head and Neck Surgery 2009;23(11):496-498
OBJECTIVE:
To investigate the indication, efficacy and influencing factors of velopharyngoplasty for treating of obstructive sleep apnea hypopnea syndrome (OSAHS).
METHOD:
Thirty OSAHS patients were included in this study, all cases had velopharyngoplasty. The follow-up period was between 6 to 11 months; and a series of measurements parameters was analysed for thirty cases.
RESULT:
When compared preoperatively and postoperatively, Apnea and hypopnea index (AHI) decreased from 54.6+/-18.3 to 25.3+/-9.5; and the Epworth sleep scale decreased from 16.5+/-4.8 to 5.2+/-4.1. According to related criterion in China, the effective rate was 70.0%.
CONCLUSION
Velopharyngoplasty can upgrade the venlopalate and enlarge the space of the oropharynx by remodeling of soft palate, contraction of scar and removal of the tonsils.
Adult
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Female
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Humans
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Male
;
Middle Aged
;
Otorhinolaryngologic Surgical Procedures
;
methods
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Palate, Soft
;
surgery
;
Sleep Apnea, Obstructive
;
surgery
10.Comparison of the effect of orplinone and Milrinone after biventricular repair of neonatal congenital heart disease
Hongjuan HUANG ; Xin LI ; Weijia SHEN ; Hongliang YUAN ; Xiaowei SHEN ; Xudong RAN ; Jianyi LIAO ; Guiying XU ; Wanyu XU
Chinese Journal of Thoracic and Cardiovascular Surgery 2023;39(11):647-652
Objective:To retrospectively compare the clinical efficacy of two PDE3 inhibitors, oplinone and Milrinone, in order to evaluate which drug has better effects on the improvement of cardiac function, protection of renal function and adverse effects of arrhythmia.Methods:A total of 41 neonates with congenital heart disease after biventricular treatment under cardiopulmonary bypass in the Department of Cardiothoracic Surgery at Soochow University Children's Hospital during 2018-2022 were collected. The experimental group was divided into two groups: Oprilinone(25 cases) and Milinone(16 cases). A retrospective study was conducted on the incidence of renal function, cardiac function improvement and arrhythmia in the children.Results:On the first day after operation, EF in both groups decreased significantly compared with that before operation( P<0.01); On day 4 after surgery, EF in the oprilinone group was significantly higher than that on day 1 after surgery( P<0.01), Milrinone group was slightly higher than that on day 1 after surgery( P<0.05), and EF in oprilinone group was significantly higher than that in Milinone group during the same period( P<0.01); EF in Milinone group continued to increase on day 7 compared with day 4( P<0.01), but there was no significant difference between the two groups. Long-term follow-up showed that there was no significant difference in EF value in the oprilinone intervention group on day 7 after surgery( P<0.05), and the long-term EF in Milinone group was higher than that at 7 days after surgery( P<0.05). The creatinine level in the oprinone intervention group continued to decrease on the 4th and 7th day after surgery( P<0.01; P<0.05); The creatinine level of Milinone group on day 4 after surgery was significantly lower than that on day 1 after surgery( P<0.01), the decrease was not significant on the 7th day after surgery compared with the 4th day after surgery; The creatinine level in the oprilinone group was lower than that in the Milrinone group on day 7 after surgery( P<0.05). The rate of arrhythmia in children was slightly decreased in the intervention group of olplinone. There was no change in the Milinone group. Conclusion:Oplinone improved cardiac function better than Milrinone, and the recovery time to normal cardiac function was shorter. In terms of renal function protection, oplinone was stronger than Milrinone, and the protective effect of oplinone on kidney lasted longer. No significant abnormalities were found with respect to adverse reactions, such as the incidence of arrhythmia.