1.Fermentative production of poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) (PHBHHx) by recombinant Aeromonas hydrophila 4AK4 (pTG01).
Shao-Ping OUYANG ; Yuan-Zheng QIU ; Qiong WU ; Guo-Qiang CHEN
Chinese Journal of Biotechnology 2003;19(6):709-714
Copolyesters consisting of 3-hydroxybutyrate (3HB) and 3-hydroxyhexanoate (3HHx) (PHBHHx), a new type of biodegradable material, are receiving considerable attentions recently. The material properties are strongly related to the 3HHx fraction of PHBHHx. As the 3HHx fraction increase, crystallinity and melting point of PHBHHx decrease, flexibility and tractility increase. PHBHHx of different 3HHx fraction can meet different demands of commercial application and research. Aeromonas are the best studied PHBHHx-producing strains. Recent studies have been focused on optimizations of fermentative culture media and culture conditions for low-cost and efficient fermentative production. Aliphatic substrates such as long-chain fatty acid and soybean oil were used in the PHBHHx fermentation as the sole carbon source and energy source. Two-stage fermentation method was also developed for more efficient PHBHHx production. While studies on Aeromonas hydrophila revealed that the monomer composition of PHBHHx could not easily be regulated by fermentative process engineering methods such as changing substrates and fermentative conditions because precursors involved in the PHBHHx synthesis were all from the beta-oxidation pathway. In this study, phbA gene encoding beta-ketothiolase and phbB gene encoding acetoacetyl-CoA reductase were introduced into a PHBHHx-producing strain Aeromonas hydrophila 4AK4 so as to provide a new 3HB precursors synthesis way. phbA gene encodes beta-ketothiolase which can catalyze two acetyl-CoA to form acetoacetyl-CoA; phbB gene encodes acetoacetyl-CoA reductase catalyzing acetoacetly-CoA into 3HB-CoA which is the precursor of 3HB. The introduced novel 3-hydroxybutyrate precursor synthesis pathway allowed the recombinant strain to use unrelated carbon source such as gluconate to provide 3HB precursors for PHBHHx synthesis. Shake-flask experiments were carried out to produce PHBHHx of controllable monomer composition and fermentations in 5 L fermentor were also proceeded for confirmation of these result in large-scale culture. In flask culture, it was possible to reduce the 3HHx mol fraction in PHBHHx from 15 % in the wild type to 3% - 12% in the recombinant by simply changing the ratio of gluconate to lauric acid in the culture media. When lauric acid was used as the sole carbon source, 51.5 g/L Cell Dry Weight (CDW) containing 62 % PHBHHx with 9.7 % 3HHx mol fraction was obtained in 56 hours of fermentation in a 5 liter fermentor. When co-substrates of sodium gluconate and lauric acid (1:1) were used as carbon sources, 32.8 g/L CDW containing 52 % PHBHHx with 6.7% 3HHx mol fraction was obtained in 48 hours of fermentation. These results showed the possibility for fermentative production of PHBHHx with controllable monomer composition.
3-Hydroxybutyric Acid
;
metabolism
;
Acetyl-CoA C-Acyltransferase
;
genetics
;
metabolism
;
Aeromonas hydrophila
;
enzymology
;
genetics
;
metabolism
;
Alcohol Oxidoreductases
;
genetics
;
metabolism
;
Bacterial Proteins
;
genetics
;
metabolism
;
Biotechnology
;
methods
;
Caproates
;
metabolism
;
Fermentation
;
genetics
;
physiology
;
Lauric Acids
;
metabolism
3.Treatment of Sepsis with Chinese Medicine: A Review Based on NF-κB Signaling Pathway
Chao HU ; Ping ZHU ; Miao JIANG ; Qian ZHANG ; Wen-xiu XU ; Li-ping OUYANG ; Xiao SHAO ; Kang SHEN ; Sai-long CHEN
Chinese Journal of Experimental Traditional Medical Formulae 2021;27(19):216-224
Sepsis, a common critical disease in the intensive care unit(ICU), features high morbidity and mortality. At present, it is mainly tackled with western medicine, which may trigger a series of problems like antibiotic resistance, adverse hormonal reactions, and high cost after a long-term use. Therefore, exploring new efficient, safe, and cheap drugs and treatment modes has become the focus of our research at this stage. By virtue of unique advantages including "the concept of holism and individualized treatment based on syndrome differentiation", Chinese medicine has accumulated quite rich experience in the prevention and treatment of sepsis. In recent years, research on the regulation of Chinese medicine on nuclear transcription factor-
4.Prognostic power of abnormal cytogenetics for multiple myeloma: a multicenter study in China.
Yue-Yun LAI ; Xiao-Jun HUANG ; Zhen CAI ; Xiang-Shan CAO ; Fang-Ping CHEN ; Xie-Qun CHEN ; Bao-An CHEN ; Mei-Yun FANG ; Jia-Fu FENG ; Wei-Ling FU ; Hai-Ying GUO ; Ming HOU ; Jian HOU ; Yu HU ; Xiao-Tong HU ; Xiao-Mei HU ; Li-Qiang HUANG ; Jie JIN ; Jian-Yong LI ; Juan LI ; Wei LI ; Ying-Min LIANG ; Ting LIU ; Qi-Fa LIU ; Yan-Hui LIU ; Ping MAO ; Jian OUYANG ; Lu-Gui QIU ; Lin QIU ; Chun-Kui SHAO ; Bin SHI ; Yong-Ping SONG ; Zi-Min SUN ; Qi-Shan WANG ; Chun WANG ; Jian-Ming WANG ; Yun-Shan WANG ; Zhao WANG ; Jian-Bo WU ; Yin-Xia WU ; Rui-Xiang XIA ; Yong-Quan XUE ; Bao-Zhen YANG ; Guang YANG ; Zheng-Lin YANG ; Li YU ; Zhong YUAN ; Sheng ZHANG ; Yin ZHANG ; Hong-Guo ZHAO ; Li ZHAO ; Dao-Bin ZHOU ; Shan-Hua ZOU ; Yun-Feng ZHU
Chinese Medical Journal 2012;125(15):2663-2670
BACKGROUNDChromosomal abnormalities have been shown to play an important prognostic role in multiple myeloma (MM). Interphase fluorescence in situ hybridization (i-FISH) has been much more effective to identify cytogenetic aberrations in MM than conventional cytogenetic technique (CC). To clearly determine the cytogenetic features of Chinese MM patients and identify their prognostic implications, we designed a multicenter study based on i-FISH including 672 patients from 52 hospitals in China.
METHODSAll 672 patients were systematically screened for the following genomic aberrations: del(13q), IgH rearrangement, del(p53) and 1q21 amplifications.
RESULTSThe analysis showed that the chromosomal changes were detected in 22.1% patients by CC and in 82.3% patients by i-FISH. The most common abnormalities by CC were chromosome 1 aberrations (48.4%), -13/13q- (37.6%), hyperdiploidy (36.6%), hypodiploidy (30.1%) and IgH rearrangements (23.7%). The most frequent abnormalities by FISH was del(13q), which was found in 60.4% patients, whereas IgH rearrangement, 1q21 amplification and p53 deletions were detected in 57.6%, 49.0% and 34.7% cases, respectively. By statistical analysis, -13/13q- by CC was associated with low level of platelet (P = 0.015), hyperdiploidy was associated with low level of serum albumin (P = 0.028), and IgH rearrangement by FISH was associated with high level of β2 microglobulin (P = 0.019). Moreover, 1q21 amplification and del(p53) by FISH conferred a high incidence of progressive disease (PD) after initial therapy. Metaphase detection of IgH rearrangements and chromosome 1 aberrations concurrently was associated with a short progression free survival (PFS) (P = 0.036). No significant prognostic implications of other cytogenetic abnormalities were found associated with overall survival and PFS.
CONCLUSIONSChinese MM patients had similar cytogenetic abnormalities compared with the previous reported studies. However, the prognostic significance of FISH aberrations were not clearly determined and further study is required.
Adult ; China ; Chromosome Aberrations ; Chromosomes, Human, Pair 1 ; genetics ; Cytogenetic Analysis ; Female ; Humans ; In Situ Hybridization, Fluorescence ; Karyotyping ; Male ; Middle Aged ; Multiple Myeloma ; genetics ; pathology
5.Recurrence Factors in Giant Cell Tumors of the Spine.
Han-Qiang OUYANG ; Liang JIANG ; Xiao-Guang LIU ; Feng WEI ; Shao-Min YANG ; Na MENG ; Ping JIANG ; Miao YU ; Feng-Liang WU ; Lei DANG ; Hua ZHOU ; Hua ZHANG ; Zhong-Jun LIU
Chinese Medical Journal 2017;130(13):1557-1563
BACKGROUNDGiant cell tumors (GCTs) are benign, locally aggressive tumors. We examined the rate of local recurrence of spinal GCTs and sought to identify recurrence factors in patients who underwent surgery.
METHODSBetween 1995 and 2014, 94 mobile spine GCT patients were treated at our hospital, comprising 43 male and 51 female patients with an average age of 33.4 years. Piecemeal intralesional spondylectomy and total en bloc spondylectomy (TES) were performed. Radiotherapy was suggested for recurrent or residual GCT cases. Since denosumab was not available before 2014 in our country, only interferon and/or zoledronic acid was suggested.
RESULTSOf the 94 patients, four underwent conservative treatment and 90 underwent operations. Seventy-five patients (79.8%) were followed up for a minimum of 24 months or until death. The median follow-up duration was 75.3 months. The overall recurrence rate was 37.3%. Ten patients (13.3%) died before the last follow-up (median: 18.5 months). Two patients (2.6%) developed osteogenic sarcoma. The local recurrence rate was 80.0% (24/30) in patients who underwent intralesional curettage, 8.8% (3/34) in patients who underwent extracapsular piecemeal spondylectomy, and 0 (0/9) in patients who underwent TES. The risk factors for local recurrence were lesions located in the cervical spine (P = 0.049), intralesional curettage (P < 0.001), repeated surgeries (P = 0.014), and malignancy (P < 0.001). Malignant transformation was a significant risk factor for death (P < 0.001).
CONCLUSIONSCervical spinal tumors, curettage, and nonintact tumors were risk factors for local recurrence. Intralesional curettage and malignancy were the most important significant factors for local recurrence and death, respectively.
6.Systematic identification of chemical forms of key terpene synthase in Cinnamomum camphora.
Qing MA ; Rui MA ; Ping SU ; Ye SHEN ; Mei-Lan CHEN ; Bao-Long JIN ; Shao-Lin OUYANG ; Juan GUO ; Guang-Hong CUI ; Lu-Qi HUANG
China Journal of Chinese Materia Medica 2023;48(9):2307-2315
Cinnamomum camphora is an important economic tree species in China. According to the type and content of main components in the volatile oil of leaf, C. camphora were divided into five chemotypes, including borneol-type, camphor-type, linalool-type, cineole-type, and nerolidol-type. Terpene synthase(TPS) is the key enzyme for the formation of these compounds. Although several key enzyme genes have been identified, the biosynthetic pathway of(+)-borneol, which has the most economic value, has not been reported. In this study, nine terpenoid synthase genes CcTPS1-CcTPS9 were cloned through transcriptome analysis of four chemical-type leaves. After the recombinant protein was induced by Escherichia coli, geranyl pyrophosphate(GPP) and farnesyl pyrophosphate(FPP) were used as substrates for enzymatic reaction, respectively. Both CcTPS1 and CcTPS9 could catalyze GPP to produce bornyl pyrophosphate, which could be hydrolyzed by phosphohydrolase to obtain(+)-borneol, and the product of(+)-borneol accounted for 0.4% and 89.3%, respectively. Both CcTPS3 and CcTPS6 could catalyze GPP to generate a single product linalool, and CcTPS6 could also react with FPP to generate nerolidol. CcTPS8 reacted with GPP to produce 1,8-cineol(30.71%). Nine terpene synthases produced 9 monoterpene and 6 sesquiterpenes. The study has identified the key enzyme genes responsible for borneol biosynthesis in C. camphora for the first time, laying a foundation for further elucidating the molecular mechanism of chemical type formation and cultivating new varieties of borneol with high yield by using bioengineering technology.
Cinnamomum camphora/enzymology*
;
Alkyl and Aryl Transferases/chemistry*
7.Chinese Trauma Surgeon Association for management guidelines of vacuum sealing drainage application in abdominal surgeries-Update and systematic review.
Yang LI ; Pei-Yuan LI ; Shi-Jing SUN ; Yuan-Zhang YAO ; Zhan-Fei LI ; Tao LIU ; Fan YANG ; Lian-Yang ZHANG ; Xiang-Jun BAI ; Jing-Shan HUO ; Wu-Bing HE ; Jun OUYANG ; Lei PENG ; Ping HU ; Yan-An ZHU ; Ping JIN ; Qi-Feng SHAO ; Yan-Feng WANG ; Rui-Wu DAI ; Pei-Yang HU ; Hai-Ming CHEN ; Ge-Fei WANG ; Yong-Gao WANG ; Hong-Xu JIN ; Chang-Ju ZHU ; Qi-Yong ZHANG ; Biao SHAO ; Xi-Guang SANG ; Chang-Lin YIN
Chinese Journal of Traumatology 2019;22(1):1-11
Vacuum sealing drainage (VSD) is frequently used in abdominal surgeries. However, relevant guidelines are rare. Chinese Trauma Surgeon Association organized a committee composed of 28 experts across China in July 2017, aiming to provide an evidence-based recommendation for the application of VSD in abdominal surgeries. Eleven questions regarding the use of VSD in abdominal surgeries were addressed: (1) which type of materials should be respectively chosen for the intraperitoneal cavity, retroperitoneal cavity and superficial incisions? (2) Can VSD be preventively used for a high-risk abdominal incision with primary suture? (3) Can VSD be used in severely contaminated/infected abdominal surgical sites? (4) Can VSD be used for temporary abdominal cavity closure under some special conditions such as severe abdominal trauma, infection, liver transplantation and intra-abdominal volume increment in abdominal compartment syndrome? (5) Can VSD be used in abdominal organ inflammation, injury, or postoperative drainage? (6) Can VSD be used in the treatment of intestinal fistula and pancreatic fistula? (7) Can VSD be used in the treatment of intra-abdominal and extra-peritoneal abscess? (8) Can VSD be used in the treatment of abdominal wall wounds, wound cavity, and defects? (9) Does VSD increase the risk of bleeding? (10) Does VSD increase the risk of intestinal wall injury? (11) Does VSD increase the risk of peritoneal adhesion? Focusing on these questions, evidence-based recommendations were given accordingly. VSD was strongly recommended regarding the questions 2-4. Weak recommendations were made regarding questions 1 and 5-11. Proper use of VSD in abdominal surgeries can lower the risk of infection in abdominal incisions with primary suture, treat severely contaminated/infected surgical sites and facilitate temporary abdominal cavity closure.
Abdomen
;
surgery
;
China
;
Drainage
;
methods
;
Evidence-Based Medicine
;
Humans
;
Practice Guidelines as Topic
;
Societies, Medical
;
organization & administration
;
Surgical Wound Infection
;
prevention & control
;
Traumatology
;
organization & administration
;
Vacuum