1.Isolation, purification and preservation of adipose-derived stem cells:research progress and future development
Youbai CHEN ; Conghui CHEN ; Zhang QIXU ; Yan HAN
Chinese Journal of Tissue Engineering Research 2016;20(10):1508-1520
BACKGROUND:In 2001, Zuk et al found adipose-derived stem cels (ASCs) from the aspirate of liposuction for the first time, which launched a new era of stem cel research. In recent years, stem cels have been proved to widely exist in many tissues and organs. ASCs are always in the spotlight of plastic and reconstructive surgery, tissue engineering and regenerative medicine because of extensive sources and simple isolation.
OBJECTIVE: To review the fat tissue harvesting and ASCs isolation, purification, expansion, and cryopreservation, to discuss the main factors which influence the yield, proliferation capacity and differentiation potential of ASCs, and to predict the future research interests based on current issues.
METHODS:On September 10th, 2015, relevant articles were searched in PubMed using the folowing format: (adipose stem cels[Title]) OR (adipose-derived stem cels[Title]) OR (adipose-derived mesenchymal stem cels[Title]) and in SinoMed using the folowing format in Chinese: (“adipose-derived stem cels” [Title])or(“adipose-derived mesenchymal stem cels”[Title]). Finaly, 81 representative articles were included according to their titles and abstracts. In this review, we also introduced relevant experience about the aforementioned procedures from the Department of Plastic Surgery and Tissue Regeneration and Molecular Cel Engineering Lab of University of Texas, MD Anderson Cancer Center, USA.
RESULTS AND CONCLUSION:The widely dispersed fat tissues potentialy provide abundant stem cels for tissue engineering and regenerative medicine. Liposuction is a mini-invasive approach for harvesting fat tissues. Colagenase digestion is the major method for ASCs isolation due to its simplicity and high yield in basic research. However, clinical fat transplantation without ASCs isolation or non-colagenase isolation of stromal vascular fraction or ASCs is preferred. The phenotype, proliferation and differentiation capacity of ASCs may be affected by several factors during the fat tissue harvesting and ASCs isolation. Therefore, a standard protocol for ASCs isolation is needed.
2.Comparisons of airway establishment among four surgical methods
Youbai CHEN ; Haizhong ZHANG ; Conghui CHEN ; Yan HAN
Chinese Journal of Emergency Medicine 2016;25(4):450-454
Objective To compare efficacy, time consumed and complications among cricothyroid membrane puncture guided tracheostomy ( CMPGT ) , surgical tracheostomy ( ST ) , surgical cricothyroidotomy ( SC) , and percutaneous tracheostomy using Griggs'guide wire dilating forceps ( GWDF) for the establishment of airway in urgent need of medical attention.Methods Twenty miniature swine were randomly ( random number) divided into four groups.The procedures of CMPGT, ST, SC and GWDF were carried out when patients'SpO2 ( oxygen saturation of blood ) declined to 80% by suspension of oxygen supply after general anesthesia. Procedure performed time, ventilation resumed time, SpO2 and electrocardiograph ( ECG) and arterial blood gases ( ABG) analysis including SaO2 , PaO2 , PaCO2 , blood pH, and heart rate, blood pressure were recorded.Fiberoptic bronchoscope was used to assess any damage to the tracheal wall.Complications were noted and scored in a two-month follow-up period.Results Airways were successfully established in all swine.The times consumed for SC, GWDF, CMPGT, and ST were (86 ±12) s, (165 ±63) s, (174 ±34) s, and (519 ±128) s, respectively, however a shorter time for ventilation resumed was found in CMPGT procedure (23 ±4) s, P<0.01.ECG showed that SpO2 and T-wave decreased and Q-T shortened after oxygen suspension and recovered to normal level rapidly after ventilation.There were significant differences in ECG and ABG between pre-and post-operative periods ( P<0.05) in all groups.Minimal intra-operative bleeding was found in two swine of each group.In ST group, moderate intra-operative bleeding was encountered in three swine.Three pigs were suffered from hypotension owing to prolonged hypoxemia.There was minimal postoperative bleeding occurred in one swine, thus leading to stoma infection.In SC group, moderate intra-operative bleeding was noticed in one swine. One miniature swine had slight injury at laryngeal cartilage resulting in difficult decannulation happened in one swine.and moderate tracheal wall injury occurred in one swine of GWDF group.The complication scores of CMPGT, GWDF, SC and ST were 3, 5, 9, and 19, respectively.Conclusions A shorter time for ventilation resumed with fewer complications was found in the procedure of CMPGT than that in the other methods.This animal study suggested that CMPGT was a time-saving and secure technique for emergency surgical airway establishment which has been practiced in human being as well.Key words:Surgical airway establishment, Cricothyroidotomy, Tracheostomy, Percutaneous dilational tracheostomy.
3.Survey of Venom Protein, Peptide and Amino Acids in Natural Drug
Jingbing CHEN ; Honglian CHEN ; Guangzhong WANG ; Xiangsheng LI ; Conghui ZHOU ;
Chinese Traditional Patent Medicine 1992;0(10):-
To review the study progresses in venom protein, peptide and amino acids in natural drug. The study progresses were reviewed on the basis of analying the collected articles. Concerning the distribution, chemical structure, property, pharmacology and toxicity of ricin, abrin, riscotoxin, snake venom, bee venom and buthotoxin. These compounds have certain toxicity and biological activity to animals, It's worth exploiting and utilizing them in conjunction with the achievement in modern chemistry and pharmacology.
4.External drainage of full pancreatic juice decreases pancreatic leakage in pancreatoduodenectomy
Yongbing CHEN ; Banglv JU ; Xiaoqing SHEN ; Conghui YU
International Journal of Surgery 2014;41(6):380-382
Objective To summarize the results of a new pancreaticojejunostomy technique designed to decrease incidence of pancreatic leakage in pancreatoduodenectomy.Methods The clinical data of 11 cases of pancreatoduodenectomy using remnant pancreatico-jejunum end-to-end anastomosis with external drainage of full pancreatic juice was analyzed retrospectively.Briefly,5-0 bioabsorbable suture was penetrated through the pancreatic stump and stent and then circulated the pancreatic stump for 2 circles and ligated.Octreotide was not administered in any cases post-operation.Results The operation was simple and timespared.Neither pancreatic leakage nor postoperative bleeding occurred in the cases.From 4 months to 3 years following operation,no other complications were found in all cases.Conclusions The method was safe,feasible and effective to decrease incidence of pancreatic leakage in pancreatoduodenectomy.
5.Application and obstacles of ANAMMOX process.
Jin RENCUN ; Zhengzhe ZHANG ; Yuxin JI ; Hui CHEN ; Qiong GUO ; Yuhuang ZHOU ; Conghui WU ; Rencun JIN
Chinese Journal of Biotechnology 2014;30(12):1804-1816
Anaerobic ammonium oxidation (ANAMMOX), as its essential advantages of high efficiency and low cost, is a promising novel biological nitrogen elimination process with attractive application prospects. Over the past two decades, many processes based on the ANAMMOX reaction have been continuously studied and applied to practical engineering, with the perspective of reaching 100 full-scale installations in operation worldwide by 2014. Our review summarizes various forms of ANAMMOX processes, including partial nitritation-ANAMMOX, completely autotrophic nitrogen removal over nitrite, oxygen limited autotrophic nitrification and denitrification, denitrifying ammonium oxidation, aerobic deammonification, simultaneous partial nitrification, ANAMMOX and denitrification, single-stage nitrogen removal using ANAMMOX and partial nitritation. We also compare the operating conditions for one-stage and two-stage processes and summarize the obstacles and countermeasures in engineering application of ANAMMOX systems, such as moving bed biofilm reactor, sequencing batch reactor and granular sludge reactor. Finally, we discuss the future research and application direction, which should focus on the optimization of operating conditions and applicability of the process to the actual wastewater, especially on automated control and the impact of special wastewater composition on process performance.
Ammonia
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chemistry
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Bioreactors
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Denitrification
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Nitrification
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Nitrites
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chemistry
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Nitrogen
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chemistry
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Oxygen
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chemistry
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Sewage
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chemistry
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Waste Disposal, Fluid
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methods
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Waste Water
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chemistry
6.Experiences of reducing complications of post-pancreaticoduodenectomy (report of 62 cases)
Conghui YU ; Hongfeng NIE ; Yongbing CHEN ; Junbo YAO ; Ronghua YANG ; Jianfei WANG
International Journal of Surgery 2012;39(2):94-97
ObjectiveTo discuss the treatment of complications post-pancreticoduedenectomy by reviewing the experiences from 62 cases.MethodsSixty-two cases of pancreticoduedenectomy from March 2001 to April 2011,mean age 59.3 ±4.1 years,were reviewed.All cases were divided into early group 24 cases and recent group (38 cases) by the difference in following aspects:perioperative management,the way of pancreatic bowel anastomosis and material choice,the vessels management,materials using in preventing leakage and bleeding.The differences in leakage,bleeding,infection,the stomach retention,biliary infections,liver abscess and incision infection in two groups were compared.ResultsThe early group showed pancreatic leakage in 10 cases,bleeding in 4 cases,celiac infection in 10 cases,gastric retention in 9 cases,biliary infections in 11 cases,liver abscess in 6 and infection of incision in 7 cases,respectively.Recent groups were in 4,0,0,0,1,2 and 3 cases,respectively.The comparisons showed the complications had reduced significantly in recent group than in the early group (x2 =77.08,P < 0.001 ).ConclusionsMinimally invasive and high quality materials use,strengthening the perioperative management are the effective measures to reduce the postoperative complications.
7.Extraction Process Optimization of Fufang Jinsha Lidan Granule by Orthogonal Experiments
Ailing YI ; Fusheng ZOU ; Bo YI ; Guang DU ; Conghui ZHOU ; Yingying WEI ; Wei CHEN
China Pharmacist 2016;19(10):1963-1965
Objective:To optimize the extraction technology of Fufang Jinsha Lidan granule. Methods:According to the properties of traditional Chinese medicines in the formula, orthogonal experiments were used to optimize the extraction process of Fufang Jinsha Li-dan granule. The volume of water, boiling time and boiling frequency were used as the three influencing factors with three different lev-els in the orthogonal experiments. Moreover, the content of paeoniflorin and the yield of extract were chosen as the evaluation indices. The orthogonal experiments were carried out according to the L9 (34 ) orthogonal table. Results:The optimal extraction process of Fu-fang Jinsha Lidan granule was as follows:boiled twice, and one hour per time with 12-fold amount of water ( soaked for 30 min with 14-fold amount of water for the first extraction process) . Conclusion:The extraction process has such properties as high extraction rate, stability, simple operation, high yield of extract and controllable quality, which is worthy of wide application.
8.Emergency treatment of pelvic fracture complicated with traumatic rupture of urethra and bladder
Jinyu LI ; Guixi CHEN ; Yansheng WU ; Zhipeng ZHENG ; Conghui SHI ; Gonglei CHEN ; Qingquan ZENG ; Zhiwei ZENG ; Chang LI
Clinical Medicine of China 2012;28(9):960-962
ObjectiveTo investigate emergency diagnosis and treatment of pelvic fracture complicated with traumatic rupture of urethra and bladder,and to improve the success rate of treatment on pelvic fracture.MethodsClinical data of 52 cases of pelvic fracture complicated with traumatic rupture of urethra and bladder in department of emergency and urology from 2000 to 2010 was retrospectively analyzed.Results Among the 52 patients,there was 41 cases of pelvic fracture complicated with posterior urethral disruption,15 cases complicated with rupture of bladder and 4 cases complicated withtraumatic rupture of urethra and bladder at the same time.In 41 cases with posterior urethral rupture,6 individual's condition were relatively so severe that they onlyunderwent bladder puncture nephrostomy,and 29 cases underwent traction urethral realignment,the other 6 cases didn't undergo surgery; In 15 cases of patients with bladder rupture,2 patients were performed urethral realignment and bladder repair,11 patients underwent the bladder repair only and the other 2 patients were not performed surgery.There were 8 patients died and the mortality rate was 15.4%.Six died cases failed to conduct emergency surgery because of uncontrollable bleeding and another 2 cases died due to multiple organ failure.ConclusionPelvic fractures is a disease with more complications,it should be diagnosed as early as possible.Patients invalid for conventional anti-shock should be performed pelvic external fixation and emergency embolization to stop bleeding in the emergency department,and undergo associated processing after they are in stable condition.
9. Application of low-depth whole-genome sequencing for copy number variations in genetic diagnosis of X-linked ichthyosis due to STS gene deletion
Zhouxian BAI ; Chen CHEN ; Lisha SU ; Hui XU ; Conghui WANG ; Panlai SHI ; Xiangdong KONG
Chinese Journal of Dermatology 2019;52(10):736-742
Objective:
To evaluate the application value and significance of low-depth whole-genome sequencing for copy number variations (CNV-Seq) in the genetic diagnosis and prenatal diagnosis of X-linked ichthyosis (XLI) due to STS gene deletion.
Methods:
Clinical data were collected from 3 616 subjects who received CNV-Seq, and single-gene test results were collected from 7 patients or pedigrees with ichthyosis in The First Affiliated Hospital of Zhengzhou University in 2018. The 3 616 samples included 2 891 prenatal samples from pregnant women (most were amniotic fluid samples, some fetal villus samples, very few umbilical blood samples) and 725 peripheral blood samples from other subjects. Genomic DNA was extracted from amniocytes or peripheral blood, and then subjected to CNV-Seq. Quantitative PCR (qPCR) and single nucleotide polymorphism (SNP) -comparative genomic hybridization (CGH) array were performed to verify the detected CNVs. Pathogenicity of the CNVs was analyzed according to the database of genomic variants (DGV) , database of genomic variation and phenotype in humans using ensembl resources (DECIPHER) , clinical genome resource (ClinGen) and online Mendelian inheritance in man (OMIM) .
Results:
Of the 3 616 subjects receiving CNV-Seq, Xp22.31 deletion was identified in prenatal samples from 6 pregnant women, including 5 male and 1 female fetuses. The deleted fragment of Xp22.31 covered the XLI region containing the major gene STS. The parental CNV-Seq showed that the Xp22.31 deletion was spontaneous mutation in 2 of the 6 fetuses, and inherited from the parents in the other 4 fetuses. qPCR confirmed that the female fetus was a carrier of a complete heterozygous deletion of the STS gene, and there was a complete deletion of the STS gene in the other 5 male fetuses. SNP-CGH array also confirmed that the female fetus was heterozygous Xp22.31 deletion carrier, which was consistent with the CNV-Seq results. Ichthyosis gene panel sequencing in the 7 patients with ichthyosis showed 1 with harlequin ichthyosis, 2 with ichthyosis vulgaris, 3 with XLI, and no causative mutation in 1. CNV-Seq confirmed that Xp22.31 deletion existed in the above 2 patients with XLI due to STS gene deletion. Moreover, Xp22.31 duplication was found in 16 out of 3 616 subjects receiving CNV-Seq, but they were all individuals or fetuses with normal phenotype.
Conclusions
CNV-Seq is a stable and reliable method for screening whole-genome CNVs, and can be applied to genetic diagnosis and prenatal diagnosis of XLI due to STS gene deletion. The deletion of Xp22.31 fragment containing the STS gene can cause XLI, and the duplication of the same region is highly likely to be the polymorphic variation.
10.Spectrum of pathological genetic variants among 405 Chinese pedigrees affected with oculocutaneous albinism.
Conghui WANG ; Chen CHEN ; Xuechao ZHAO ; Ganye ZHAO ; Lina LIU ; Xiangdong KONG
Chinese Journal of Medical Genetics 2020;37(7):725-730
OBJECTIVE:
To determine the spectrum of pathological genetic variants among 405 Chinese pedigrees affected with oculocutaneous albinism (OCA).
METHODS:
A total of 405 OCA patients were collected. High-throughput sequencing (The panel included TYR, OCA2, TYRP1 and SLC45A2 genes), Sanger sequencing and multiplex ligation-dependent probe amplification (MLPA) were used to analyze the genetic variants and patterns of each subtype.
RESULTS:
The overall detection rate of genetic variants was 79.9% (647/810), and the variants included missense variants (57.3%, 371/647), frameshift variants (22.9%, 148/647), nonsense variants (13.9%, 90/647), splicing variants (5.6%, 36/647), and microdeletions (0.3%, 2/647). Thirty-six novel variants were detected. Of the 405 patients, 306 have carried 2 variant alleles (75.6%, 306/405), 35 carried 1 variant alleles (8.6%, 35/405), while no variant was detected in 64 patients. Among the 306 genetically diagnosed OCA patients, OCA1 was the most common form (74.5%, 228/306), compared with OCA2 (15.0%, 46/306), OCA3 (0.7%, 2/306) and OCA4 (9.8%, 30/306), respectively. One patient was found to harbor homozygous c.1262-4_c.1262-3insTAGA variant of the TYRP1 gene. Another patient was found to carry compound heterozygous variants of c.1214C>A (p.T405N) and c.1338delinsCG(p.V447Gfs*19) of the TYRP1 gene.
CONCLUSION
High-throughput sequencing in combination with Sanger sequencing and MLPA can effectively detect genetic variants associated with OCA. Above finding has expanded variant spectrum of OCA, which can facilitate genetic and prenatal diagnosis of this disease in China.