1.The oxidative modification of albumin by hydrochlorous acid and the relationship between the oxidized HSA and AOPPs
Yanwen LU ; Qiuyu ZHU ; Feng DING ; Yong GU ; Shanyan LIN
Chinese Journal of Nephrology 2005;0(08):-
Objective To study the effect of oxidative modification of hydrochlorous acid (HOCl) on human serum albumin (HSA) and the relationship between the AOPPs and HOCl-treated HSA. Methods Purified HSA (60 mg/ml) was treated with HOCl (0, 1, 5, 10, 20, 30, 40, 50 and 60 mmol/L). Size-exclusion chromatography was applied to estimate molecular weights of oxidized products of HSA by HOCl and spectrum scan from 190 nm -400 nm was performed to observe the spectrum characteristics of all variants of HSA. Results Major products of HSA after exposure to HOC1 were dimer and hexmer of HSA. The first-order process could be employed to describe the oxidative dynamics of monomer and dimer of HSA oxidized by HOCl. To AOPPs formation mediated by oxidant was identified as pseudo first-order reaction. However, formation hexmer was much in accordance with second-order reaction. Hexmer was also a major contributor to AOPPs in all types of modified HSA. Spectral analysis showed that red shift of absorbance maximum of polymers of HSA occurred, suggesting that a possibility that polymers of HSA were cross linked by tyrosine residues in protein. Conclusions Protein aggregation is primary consequence of HSA after its exposure to HOCl. Hexmer of HSA is the major contributor to AOPPs.
2.Formation of advanced oxidative protein products induced by three uremic toxins
Yanwen LU ; Qiuyu ZHU ; Feng DING ; Yong GU ; Shantan LIN
Chinese Journal of Nephrology 2008;24(6):435-440
Objective To study whether the uremic toxins accumulated long-term in uremia patients may be involved in oxidation of protein by forming advanced oxidative protein products (AOPPs). Methods Malonylaldehyde (MDA), hippuric acid (HA) and p-cresol were used as the representatives of uremic toxins. Human albumin serum (HSA), plasma specimens from normal or uremia patients were incubated respectively with MDA (10 retool/L), HA (20 mmol/L) and p-cresol (10 retool/L) or PBS (20 retool/L, pH 7.4, as control groups) at 37℃ for 30 minutes or 24 hours, respectively. Those indices such as AOPPs, protein thiol groups (Pt-SH) and dityrosine were used as biomarkers of protein injury. High performance liquid chromatography (HPLC) was employed to identify the aggregation and cross-links of modified proteins. Results AOPPs levels in all groups containing poison compounds were significantly increased by 121.5%(P<0.05) compared to that in control groups. Uremic toxins also resulted in over 14.7% loss in Pt-SH (P< 0.05) and 119.2% increment in dityrosine, respectively (P<0.05). Meanwhile, the formation of HMW-AOPPs in a time-dependent manner was observed by HPLC and cross-linked protein levels were significantly increased by 148.45%~333.3% in comparison with control groups. Conclusion Uremic toxins can directly mediate the damage of proteins by inducing the formation of HMW- AOPPs in a time-dependent manner, which is also one of the mechanism of AOPPs production in vivo besides the activation of the myeloperoxidase-H2O2-Cl pathway.
3.Cysteine and arginine in plasma albumin are the major targets attacked by free radicals
Yanwen LU ; Qiuyu ZHU ; Feng DING ; Yong GU ; Shanyan LIN
Chinese Journal of Nephrology 1997;0(01):-
Objective To study the effects of oxidants on the structure of albumin. Methods Using both AOPPs and protein carbonyl content as indices. The oxidative stress level in normal controls and uremia patients was evaluated. Albumin in plasma was purified by HPLC and then was subjected to amino acids composition assay. Results Both AOPPs level and protein carbonyl content in uremic patients were significantly higher than those in controls (P
4.Generation of hematopoietic stem/progenitor cells with property of strengthened cell mediated immunity from an embryonic stem cell line
Qifeng ZHOU ; Yanwen PENG ; Lianqiang FENG ; Xiuming ZHANG ; Yan LI ; Shunon LI
Chinese Journal of Pathophysiology 2000;0(10):-
AIM: To induce lymphoid stem cells and/or T-cell precursors to diffe rentiate into functional mature T lymphocyte, and to increase the surface marker of T lymphocytes such as CD + 3, while embryonic stem(ES) cells differentiate d into hematopoietic stem/progenitor cells(HSPCs) in vitro . When they were i njec ted into lethally irradiated mice, these differentiated cells had the advantage in immune reconstitution. METHODS: Embryonic stem cells formed e mbryoid bodies(EBs) in the medium containing methycellulose, hematopoietic growt h factors(HGFs) was added to the culture system on the 6th day, thymopeptide was added at the same time. Flow cytometry were performed to detect the surface mar ker CD 34 and CD 3 of the differentiated cells. Finally the differentiated cells were injected into lethally irradiated mice, 60 days later, the incidence rate of graft versus host disease(GVHD) was taken as the mark of cell mediated immunity, PCR was performed to detect the sex determining region of the Y-chromo some(Sry) in bone marrow cells and spleen cells of the survival host female mice . RESULTS: The percentage of CD + 3 T lymphocytes was 10.52% a nd the incidence rate of GVHD was 0% on the 13th day, but they respectively rose up to 22.93% an d 100% if thymopeptide was added in the procedure of inducing ES cells to differ entiate into HSPC in vitro . CONCLUSION: The quantity of CD + 3 T lymphocytes increased in medium containing thymopeptide when ES cells differe ntiated into CD 34 + HSPC.
5.Pedicled muscular flap for treatment of pyothorax-resulted wound.
Yong ZHANG ; Zihao FENG ; Yanwen YANG ; Chunlai LU ; Di GE ; Fazhi QI
Chinese Journal of Plastic Surgery 2014;30(6):428-431
OBJECTIVETo evaluate the therapeutic effect of pedicled muscular flaps combined with partial thoracotomy for treatment of pythorax-resulted wounds and defects.
METHODS35 cases with pythorax-resulted refractory infected wound and dead space, were retrospectively analyzed. The wound and dead space were eliminated with pedicled muscular flaps, including latissimus dorsi muscular flaps and serratus anterior muscle flaps in 21 patients, pectoralis major muscle flaps with rectus muscle flaps in 8 patients, latissimus dorsi muscul flaps and vertical rectus muscle flaps in other 6 patients.
RESULTS34 flaps survived completely except for 1 verticle rectus muscular flap with partial necrosis at distal end. Primary healing was achieved in 29 cases. The other 6 cases were discharged with drainage. Among the 6 cases, 3 cases healed spontaneously, 2 cases underwent reoperation, 1 case need long-term drainage in the fistula.
CONCLUSIONPedicled muscular flap, combined with partial thoracotomy can effectively treat refractory wound and dead space resulted by pythorax.
Combined Modality Therapy ; methods ; Empyema, Pleural ; surgery ; Humans ; Myocutaneous Flap ; transplantation ; Pectoralis Muscles ; transplantation ; Reoperation ; Retrospective Studies ; Superficial Back Muscles ; transplantation ; Thoracotomy ; methods ; Wound Healing
6.Levels of autophagy in T cells and B cell of patients with systemic lupus erythe-matosus and its clinical significance
Yanwen CHEN ; Chao WANG ; Xuguang WANG ; Yongmin FENG ; Ning AN ; Hongluan WU ; Shangmei LI ; Jingli TAO ; Weijing LIU ; Huafeng LIU ; Qingjun PAN
Chinese Journal of Immunology 2015;(10):1380-1384,1388
Objective:To investigate levels of autophagy in T cells and B cell of patients with systemic lupus erythematosus ( SLE) and its clinical significance.Methods: 68 SLE patients without treatment within 4 weeks were enrolled in this study.We accessed the levels of autophagy in T cells and B cells of 23 healthy controls and 68 patients before and after treatment by flow cytometry,and analyzed their correlations with serum levels of C3 and anti-dsDNA antibodies,SLEDAI score,et al.Results: Before treatment,a significantly increased levels of LC3-Ⅱ was observed in SLE patients than healthy controls, the active group ( SLEDAI score≥10) was significantly higher than the stable group(SLEDAI score<10),and the newly diagnosed group was significantly higher than the recurrent group(all P<0.05).While the levels of LC3-Ⅱ was decreased in B lymphocytes in SLE patients,the active group was significantly lower than stable group,and the active group was significantly lower than the newly diagnosed group(all P<0.05). Correlation analysis found that,a positively correlation was observed for the levels of LC3-Ⅱwith SLEDAI score in T lymphocyte( rs=0.289,P<0.05),and the levels of C3 were positively correlated the levels of LC3-Ⅱ in B lymphocyte(rs=0.371,P<0.01).After treatment for five days, levels of autophagy in T lymphocytes of SLE patients with good prognosis ( SLEDAI score decreased ≥4 ) significantly decreased(P<0.05).Also,three days after treatment,levels of autophagy in B lymphocytes of SLE patients with good prognosis were increased significantly ( P<0.05 ) .However, SLE patients with poor prognosis had no such difference ( P>0.05 ) . Conclusion:Levels of autophagy in T and B lymphocytes of SLE patients are abnormal compared to healthy controls,and these changes are associated with disease activity.Also,these changes are expected to be the indicators of disease activity and potential therapeutic targets in SLE.
7.Xenogenic acellular dermal matrix in immediate implant-based breast reconstruction following nipple/skin-sparing mastectomy
Zihao FENG ; Yong ZHANG ; Yanwen YANG ; Cheng CHEN ; Fazhi QI
Chinese Journal of Medical Aesthetics and Cosmetology 2018;24(1):19-23
Objective To explore the clinical outcomes of xenogenic acellular dermal matrix in immediate implant-based breast reconstruction following nipple/skin sparing mastectomy.Methods From January 2015 to June 2017,patients that underwent immediate implant-based breast reconstruction following nipple/skin-sparing mastectomy were retrospectively studied.Results 43 female patients with breast cancer were enrolled.A retrospective review was performed on patients that were reconstructed with either alloplastic reconstruction (group A,18 breasts) or xenogenic acellular matrix dermal (group B,25 groups).Differences in the average age,body mass index,numbers of diabetic patients,numbers of active smoker,and volume of implant between the two groups were not statistically significant (P>0.05).All patients were followed up from 3 to 28 months,with the middle time of 18,7 months.Complications in both groups of patients were clinically recorded after operation.When comparing clinical outcomes,group A had fewer drain (523.17±62.18) ml vs (639.56±54.34)ml,and fewer days before drain removal (7.83±1.58) d vs (10.2±1.44) d.No significant difference was seen in terms of seroma,hematoma,delayed healing,infection,nipple necrosis,capsular contracture,or tumor recurrence.Conclusions The use of xenogenic acellular matrix dermal in immediate implant-based breast reconstruction following nipple/skin-sparing mastectomy is as safe and effective as alloplastic reconstruction.
8.Vertical reduction mammoplasty based on Würinger's horizontal septum
Jianying GU ; Yong ZHANG ; Zihao FENG ; Yanwen YANG ; Yuedong SHI ; Fazhi QI
Chinese Journal of Medical Aesthetics and Cosmetology 2018;24(1):24-27
Objective The vertical mammoplasty which improves the breast morphology and reduces the operative scar has become the first selection of breast reduction.However,the nipple-and-areola complex (NAC) had the risk of necrosis in those severe patients.In this report,we performed the vertical reduction mammoplasty based on Wu'ringer's horizontal septum to reduce the NAC necrosis possibility.Methods Typical Lejourmosque-dome design was performed.The inferior part of glandular tissue and the skin were excised.The NAC was elevated to normal position based on Wu'ringer's horizontal septum.The breast morphology was modified with vertical scar left.Results 79 patients were operated.4 patients had unilateral breast reduction.4 breasts had partial wound dehiscence.They were treated conservatively with topical dressing.No complete NAC necrosis occurred.Conclusions The vertical reduction mammoplasty based on Wu'ringer's horizontal septum is a safe and effective procedure.The NAC necrosis risk is reduced.This method deserves to be recommended.
9.Therapeutic evaluation of surgical procedure in treatment of secondary hyperparathyroidism.
Jialin FENG ; Qinyi ZHOU ; Jun CHEN ; Yanwen LUO ; Jiadong WANG
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2015;50(8):661-667
OBJECTIVETo analyze the short-term and long-term outcomes of the three operation methods in treatment of secondary hyperparathyroidism.
METHODSClinical data of 88 patients who underwent parathyroidectomy for secondary hyperparathyroidism from October 2004 to October 2014 were reviewed retrospectively. Patients were divided into three subgroups, including subtotal parathyroidectomy (group I), total parathyroidectomy with autotransplantation (group II) and intraoperative ⁹⁹Tc(m)-MIBI radioguided total parathyroidectomy with autotransplantation (group III). Their serum calcium, phosphorus, serum iPTH results in the pre-operation, short-term (≤ 6 months) and long term (> 6 months) of post-operation were collected and compared. The improvements of clinical symptoms together with the postoperative recurrence rate and the complication data were observed and analyzed.
RESULTSThe symptoms of all patients were improved after the operation.The short-term postoperative serum calcium, serum iPTH and recurrence rate of Group I and group II were all not statistically significant. The short-term postoperative serum calcium, iPTH and recurrence rate of group III were significantly lower than those of group I and group II (P < 0.05). Postoperative serum phosphorus values of the three groups had no statistical difference. The operation time of group III was significantly shorter than that of group II ((77 ± 13) vs (108 ± 17) min, P < 0.05). The positive rates were more accurate in group III than in group II (98.5% vs 88.7%, P < 0.05). For the long term follow-up, the postoperative serum calcium, phosphorus, serum iPTH and recurrence rate of group I and group II were not statistically different. The postoperative recurrence rate of group III was lower than that of group I and group II (3.6% vs 31.6% and 21.4% respectively, P < 0.05).
CONCLUSIONSTotal parathyroidectomy with autotransplantation has a good efficacy to treat secondary hyperparathyroidism. The intraoperative ⁹⁹Tc(m)-MIBI radioguided total parathyroidectomy with autotransplantation can find the ectopic and supernumerary glands to improve the success rate of operation. And it also can permit omission of frozen section to reduce the operative time.In the postoperative follow-up, the recurrence rate is low, so it is a safe and effective treatment.
Calcium ; blood ; Humans ; Hyperparathyroidism, Secondary ; surgery ; Parathyroid Glands ; surgery ; Parathyroid Hormone ; blood ; Parathyroidectomy ; methods ; Phosphorus ; blood ; Postoperative Period ; Recurrence ; Retrospective Studies ; Transplantation, Autologous ; Treatment Outcome
10.Development of a multiplex qRT-PCR assay for detection of African swine fever virus, classical swine fever virus and porcine reproductive and respiratory syndrome virus
Yating CHEN ; Kaichuang SHI ; Huixin LIU ; Yanwen YIN ; Jing ZHAO ; Feng LONG ; Wenjun LU ; Hongbin SI
Journal of Veterinary Science 2021;22(6):e87-
Background:
African swine fever virus (ASFV), classical swine fever virus (CSFV), and porcine reproductive and respiratory syndrome virus (PRRSV) are still prevalent in many regions of China. Co-infections make it difficult to distinguish their clinical symptoms and pathological changes. Therefore, a rapid and specific method is needed for the differential detection of these pathogens.
Objectives:
The aim of this study was to develop a multiplex real-time quantitative reverse transcription polymerase chain reaction (multiplex qRT-PCR) for the simultaneous differential detection of ASFV, CSFV, and PRRSV.
Methods:
Three pairs of primers and TaqMan probes targeting the ASFV p72 gene, CSFV 5′untranslated region, and PRRSV ORF7 gene were designed. After optimizing the reaction conditions, including the annealing temperature, primer concentration, and probe concentration, multiplex qRT-PCR for simultaneous and differential detection of ASFV, CSFV, and PRRSV was developed. Subsequently, 1,143 clinical samples were detected to verify the practicality of the assay.
Results:
The multiplex qRT-PCR assay could specifically and simultaneously detect the ASFV, CSFV, and PRRSV with a detection limit of 1.78 × 10 0 copies for the ASFV, CSFV, and PRRSV, but could not amplify the other major porcine viruses, such as pseudorabies virus, porcine circovirus type 1 (PCV1), PCV2, PCV3, foot-and-mouth disease virus, porcine parvovirus, atypical porcine pestivirus, and Senecavirus A. The assay had good repeatability with coefficients of variation of intra- and inter-assay of less than 1.2%. Finally, the assay was used to detect 1,143 clinical samples to evaluate its practicality in the field. The positive rates of ASFV, CSFV, and PRRSV were 25.63%, 9.36%, and 17.50%, respectively. The co-infection rates of ASFV+CSFV, ASFV+PRRSV, CSFV+PRRSV, and ASFV+CSFV+PRRSV were 2.45%, 2.36%, 1.57%, and 0.17%, respectively.
Conclusions
The multiplex qRT-PCR developed in this study could provide a rapid, sensitive, specific diagnostic tool for the simultaneous and differential detection of ASFV, CSFV, and PRRSV.