1.Effect of Xidi Liangxue recipe on the proliferation and apoptosis of HaCaT cells through the lncRNA NEAT1/miR-485-5p/STAT3 regulatory network
Zhiming TANG ; Mengqing JING ; Lu LU ; Xiao SHAN ; Cuixia ZHANG ; Xiaoyu ZHANG ; Sa MENG
Chinese Journal of Dermatology 2023;56(7):642-650
Objective:To investigate the effect of Xidi Liangxue recipe on the proliferation and apoptosis of HaCaT cells through the long non-coding RNA (lncRNA) nuclear-enriched abundant transcript 1 (NEAT1) /microRNA (miR) -485-5p/signal transducer and activator of transcription 3 (STAT3) regulatory network. Methods:HaCaT cells were induced by interleukin-17 (IL-17), and the mRNA and protein expression of lncRNA NEAT1, miR-485-5p and STAT3 was detected in IL-17-induced HaCaT cells and normal human epidermal keratinocytes (NHEK) by quantitative PCR (qPCR) and Western blot analysis, respectively. The location of lncRNA NEAT1 and miR-485-5p in IL-17-induced HaCaT cells was observed by fluorescence in situ hybridization (FISH), and the targeted regulatory relationship among lncRNA NEAT1, miR-485-5p and STAT3 was verified by double-luciferase reporter gene assay. Chinese herbs were decocted according to the Xidi Liangxue recipe, SD rats were divided into two groups to be gavaged with the above decoctions (medicated group) or physiological saline (control group) for 5 days, and then serum samples were collected from the above two groups of rats separately. The IL-17-induced HaCaT cells were divided into 4 groups: control group treated with the control sera, lncRNA-NEAT1 overexpression group transfected with lncRNA-NEAT1 overexpression vectors and treated with the control sera, Xidi Liangxue recipe group treated with the medicated sera, and Xidi Liangxue recipe + lncRNA-NEAT1 overexpression group transfected with lncRNA-NEAT1 overexpression vectors and treated with the medicated sera. qPCR, Western blot analysis, flow cytometry, and cell counting kit (CCK8) assay were performed to determine the mRNA and protein expression of lncRNA NEAT1, miR-485-5p and STAT3, and to evaluate cell proliferation and apoptosis. The two independent samples t-test was used for comparisons between two groups, one-way analysis of variance for comparisons among multiple groups, and least significant difference (LSD) t-test for multiple comparisons. Results:The IL-17-induced HaCaT cell group showed significantly increased relative expression levels of lncRNA NEAT1 and STAT3 mRNA (1.84 ± 0.21, 2.20 ± 0.24, respectively) and significantly increased protein expression of STAT3 and p-STAT3 (1.27 ± 0.13, 2.43 ± 0.16, respectively), but significantly decreased expression level of miR-485-5p (0.32 ± 0.04) compared with the NHEK group (lncRNA NEAT1 and STAT3 mRNA: 1.00 ± 0.11, 1.00 ± 0.11, respectively, both P < 0.05; STAT3 and p-STAT3 protein: 1.00 ± 0.11, 1.00 ± 0.10, t = 2.54, 3.02, respectively, both P < 0.05; miR-485-5p: 1.00 ± 0.12, t = 2.94, P = 0.015). FISH demonstrated that miR-485-5p and lncRNA NEAT1 were co-located in the cytoplasm of HaCaT cells. The double-luciferase reporter gene assay showed that the relative activity of luciferase was significantly lower in the miR-485-5p group than in the negative control group (both P < 0.05) after the transfection with wild-type lncRNA NEAT1 or STAT3 recombinant plasmids, while there were no significant differences between the miR-485-5p group and negative control group after the transfection with mutant lncRNA NEAT1 or STAT3 recombinant plasmids (both P > 0.05). Compared with the control group, the lncRNA-NEAT1 overexpression group showed significantly increased expression of lncRNA NEAT1 and STAT3 (including STAT3 mRNA, STAT3 protein, and p-STAT3 protein) in HaCaT cells (all P < 0.05), but significantly decreased miR-485-5p expression ( P < 0.05) ; the Xidi Liangxue recipe group showed significantly decreased expression of lncRNA NEAT1 and STAT3 (all P < 0.05), but significantly increased miR-485-5p expression compared with the control group ( P < 0.05) ; significantly decreased expression of lncRNA NEAT1 and STAT3, but significantly increased miR-485-5p expression was observed in the Xidi Liangxue recipe + lncRNA-NEAT1 overexpression group compared with the lncRNA-NEAT1 overexpression group (all P < 0.05). After 24-, 48-, and 72-hour intervention, CCK8 assay showed that the proliferative activity of HaCaT cells was significantly higher in the lncRNA-NEAT1 overexpression group than in the control group (all P < 0.05), as well as in the Xidi Liangxue recipe + lncRNA-NEAT1 overexpression group than in the Xidi Liangxue recipe group (all P < 0.05), and the cellular proliferative activity was significantly lower in the Xidi Liangxue recipe + lncRNA-NEAT1 overexpression group and Xidi Liangxue recipe group than in the control group (all P < 0.05). The apoptosis rate was significantly lower in the lncRNA-NEAT1 overexpression group (5.84% ± 0.28%) than in the control group (14.75% ± 0.83%, LSD- t = 3.48, P = 0.002), but significantly higher in the Xidi Liangxue recipe group (35.72% ± 3.62%) than in the control group (LSD- t = 5.34, P = 0.001) ; the Xidi Liangxue recipe + lncRNA-NEAT1 overexpression group showed significantly increased apoptosis rate (27.64% ± 2.82%) compared with the lncRNA-NEAT1 overexpression group (LSD- t = 9.06, P < 0.001) . Conclusion:The Xidi Liangxue recipe could inhibit the proliferation of IL-17-induced HaCaT cells and promote their apoptosis, which may be related to the intervention in the lncRNA NEAT1/miR-485-5p/STAT3 regulatory network.
2.Efficient biosynthesis of γ-aminobutyric acid by rationally engineering the catalytic pH range of a glutamate decarboxylase from Lactobacillus plantarum.
Jiewen XIAO ; Jin HAN ; Zhina QIAO ; Guodong ZHANG ; Wujun HUANG ; Kai QIAN ; Meijuan XU ; Xian ZHANG ; Taowei YANG ; Zhiming RAO
Chinese Journal of Biotechnology 2023;39(6):2108-2125
γ-aminobutyric acid can be produced by a one-step enzymatic reaction catalyzed by glutamic acid decarboxylase. The reaction system is simple and environmentally friendly. However, the majority of GAD enzymes catalyze the reaction under acidic pH at a relatively narrow range. Thus, inorganic salts are usually needed to maintain the optimal catalytic environment, which adds additional components to the reaction system. In addition, the pH of solution will gradually rise along with the production of γ-aminobutyric acid, which is not conducive for GAD to function continuously. In this study, we cloned the glutamate decarboxylase LpGAD from a Lactobacillus plantarum capable of efficiently producing γ-aminobutyric acid, and rationally engineered the catalytic pH range of LpGAD based on surface charge. A triple point mutant LpGADS24R/D88R/Y309K was obtained from different combinations of 9 point mutations. The enzyme activity at pH 6.0 was 1.68 times of that of the wild type, suggesting the catalytic pH range of the mutant was widened, and the possible mechanism underpinning this increase was discussed through kinetic simulation. Furthermore, we overexpressed the Lpgad and LpgadS24R/D88R/Y309K genes in Corynebacterium glutamicum E01 and optimized the transformation conditions. An optimized whole cell transformation process was conducted under 40 ℃, cell mass (OD600) 20, 100 g/L l-glutamic acid substrate and 100 μmol/L pyridoxal 5-phosphate. The γ-aminobutyric acid titer of the recombinant strain reached 402.8 g/L in a fed-batch reaction carried out in a 5 L fermenter without adjusting pH, which was 1.63 times higher than that of the control. This study expanded the catalytic pH range of and increased the enzyme activity of LpGAD. The improved production efficiency of γ-aminobutyric acid may facilitate its large-scale production.
Glutamate Decarboxylase/genetics*
;
Lactobacillus plantarum/genetics*
;
Catalysis
;
gamma-Aminobutyric Acid
;
Hydrogen-Ion Concentration
;
Glutamic Acid
3.Outline of 2023 updated International Society for Peritoneal Dialysis catheter-related infection recommendations
Xiao YANG ; Na TIAN ; Wanhong LU ; Jin CHEN ; Zhiming YE ; Jie DONG ; Kai-Ming CHOW ; Kam-tao Philip LI
Chinese Journal of Nephrology 2023;39(12):964-968
Peritoneal dialysis (PD) catheter-related infections are important risk factors for catheter loss and peritonitis. 2023 International Society for Peritoneal Dialysis (ISPD) catheter-related infection recommendations have revised and clarified definitions and classifications of exit site infection and tunnel infection, such as cause-specific catheter-related infection, culture-negative catheter-related infection, refractory catheter-related infection, and infection- related catheter removal. A new target for the exit site infection rate should not exceed 0.40 episodes per year at risk. The recommendation about topical antibiotic cream or ointment to catheter exit site has been downgraded. New recommendations for exit site infection include clarified suggestion of exit site dressing cover and revised topical antibacterial agents as well as antibiotics treatment duration. In addition to catheter removal and reinsertion, new salvage options for catheter are suggested. The paper outlines the updated main content of the guide.
4.Comparative Genomics Reveals Evolutionary Drivers of Sessile Life and Left-right Shell Asymmetry in Bivalves
Zhang YANG ; Mao FAN ; Xiao SHU ; Yu HAIYAN ; Xiang ZHIMING ; Xu FEI ; Li JUN ; Wang LILI ; Xiong YUANYAN ; Chen MENGQIU ; Bao YONGBO ; Deng YUEWEN ; Huo QUAN ; Zhang LVPING ; Liu WENGUANG ; Li XUMING ; Ma HAITAO ; Zhang YUEHUAN ; Mu XIYU ; Liu MIN ; Zheng HONGKUN ; Wong NAI-KEI ; Yu ZINIU
Genomics, Proteomics & Bioinformatics 2022;(6):1078-1091
Bivalves are species-rich mollusks with prominent protective roles in coastal ecosystems.Across these ancient lineages,colony-founding larvae anchor themselves either by byssus produc-tion or by cemented attachment.The latter mode of sessile life is strongly molded by left-right shell asymmetry during larval development of Ostreoida oysters such as Crassostrea hongkongensis.Here,we sequenced the genome of C.hongkongensis in high resolution and compared it to reference bivalve genomes to unveil genomic determinants driving cemented attachment and shell asymmetry.Importantly,loss of the homeobox gene Antennapedia(Antp)and broad expansion of lineage-specific extracellular gene families are implicated in a shift from byssal to cemented attachment in bivalves.Comparative transcriptomic analysis shows a conspicuous divergence between left-right asymmetrical C.hongkongensis and symmetrical Pinctada fucata in their expression profiles.Especially,a couple of orthologous transcription factor genes and lineage-specific shell-related gene families including that encoding tyrosinases are elevated,and may cooperatively govern asymmet-rical shell formation in Ostreoida oysters.
5.pH-sensitive and bubble-generating mesoporous silica-based nanoparticles for enhanced tumor combination therapy.
Zhiming ZHANG ; Chenlu HUANG ; Li ZHANG ; Qing GUO ; Yu QIN ; Fan FAN ; Boxuan LI ; Bao XIAO ; Dunwan ZHU ; Linhua ZHANG
Acta Pharmaceutica Sinica B 2021;11(2):520-533
Chemotherapy has been a major option in clinic treatment of malignant tumors. However, single chemotherapy faces some drawbacks, such as multidrug resistance, severe side effects, which hinder its clinic application in tumor treatment. Multifunctional nanoparticles loading with chemotherapeutic agent and photosensitizer could be a promising way to efficiently conduct tumor combination therapy. In the current study, a novel pH-sensitive and bubble-generating mesoporous silica-based drug delivery system (denoted as M(a)D@PI-PEG-RGD) was constructed. Ammonium bicarbonate (NH
6.Prevalence of injury among children in 28 counties of 6 provinces in China in 2017
Ye JIN ; Pengpeng YE ; Xiao DENG ; Yuan WANG ; Yuliang ER ; Cuirong JI ; Zhiming LU ; Leilei DUAN
Chinese Journal of Epidemiology 2021;42(8):1392-1400
Objective:To investigate the incidence of injuries among children aged 0-17 years from 28 counties in six provinces.Methods:According to the earlier study on data in different local areas and the current work plan, we selected the survey objects for a general survey on child injury prevalence in the last 12 months. The subjects included guardians of children who did not attend kindergartens, children from kindergartens, and primary school grades 1-3. Children from grades 4-6 primary school, junior high school, and senior high school were also included.Results:The overall rate of injured children was 4.83%, with the incidence of injuries as 5.75% in the last 12 months before the survey. Both the rate of injured children and incidence of injuries were higher in boys than that in girls. Children whose primary caregivers were relatives, grandparents, minors, or with an education level below the primary school had a relatively higher rate of both injured children and incidence of injuries. The first cause of child injury was fall. The time of child injury was mainly from June to October, mainly at home, with the main activity as playing. The significant injuries in children were contusion and abrasion, and the main parts injured were lower limbs, upper limbs, and head. Most of the injuries were cured, and the primary treatment was outpatient and emergency treatment.Conclusions:Boys and children whose parents can not take care of are the priority population for child injury prevention and control. Targeted prevention and control measures should be implemented according to the characteristics of injuries among children of different ages.
7.The epidemiological characteristics and influencing factors of animal injuries among children from 28 counties in 6 provinces, China
Zhiming LU ; Xiao DENG ; Yuan WANG ; Yuliang ER ; Pengpeng YE ; Ye JIN ; Cuirong JI ; Leilei DUAN
Chinese Journal of Epidemiology 2021;42(8):1401-1406
Objective:To understand the epidemiological characteristic and influencing factors of animal injuries among children from 28 counties/districts in 6 provinces of China to provide evidence for formulating related prevention measures and strategies.Methods:Data was obtained from 'Children injury prevention project, 2016-2020'. Variables on the characteristics, including incidence, locations, related activities, time, and outcomes among children, were described. The negative binomial regression model analyzed animal injury frequency influencing factors.Results:There were 107 029 boys and 97 599 girls, with a gender ratio of 1.10, among the 204 628 children aged 0-17 in 28 counties/districts in 6 provinces of China. Per-person and per-time incidence rates of animal injuries were 0.70% and 0.72%, both higher in boys than in girls. Animal injuries among children mainly occurred in home (73.20%), road/street (7.51%), and open space of community/village (6.77%), during playing (63.15%), with the peak time between 12:01-18:00 (45.03%). Upper limbs (47.89%), lower limbs (35.31%), and head (8.44%) were the most injured body parts. Outpatient or emergency treatment (85.19%) had been carried out. The negative binomial regression showed that sex, grade, guardian educational level, urban or rural, and daily communication between guardians and children as influencing factors in the frequency of animal injuries among children.Conclusions:Special attention should be given to boys, at home, in projects on animal injury prevention and control. There were many influencing factors of animal injury frequency. Knowledge of injury prevention should be popularized to children and guardians. Related comprehensive intervention should be developed in families, schools, and social settings.
8.Prevalence and prevention of traffic injury among children in 28 counties of 6 provinces in China, 2017
Cuirong JI ; Ye JIN ; Pengpeng YE ; Yuan WANG ; Yuliang ER ; Xiao DENG ; Zhiming LU ; Leilei DUAN
Chinese Journal of Epidemiology 2021;42(8):1407-1412
Objective:To investigate the incidence of traffic injuries among children aged 0-17 years in 28 counties of 6 provinces and to provide prevention suggestions.Methods:Using the general survey method, a questionnaire survey was conducted on the road traffic injuries of children in 28 counties of 6 provinces in 2017.Results:A total of 204 628 students were surveyed, 908 of whom had experiences of road traffic injuries. Road traffic injuries were 0.44% among children in 28 counties of 6 provinces in China and 0.94% among children who did not attend kindergartens. July is a high incidence period of road traffic injuries among children (12.92%). When children suffer from road traffic injuries, the first three activities were playing (21.61%), riding an electric bicycle (20.40%), walking (20.07%). Most injuries appeared as contusion/abrasion, accounting for 42.26% of the total cases. The injured parts were mainly lower limbs, upper limbs, and heads, accounting for 38.85%, 23.29%, and 20.20%. Most children with road traffic injuries recovered after related treatments, accounting for 85.43% of all cases. The median length of hospital stay of children with road traffic injuries was seven days, and the median expense in hospitals was 3 000 Yuan (RMB).Conclusions:The incidence of road traffic injuries among males appeared higher than females, and the children who did not attend the kindergartens were the highest. During playing, riding an electric bicycle and walking were the top three activities prone to road traffic injuries.
9. Investigation and analysis on fingertip vibrotactile perception threshold at two frequencies in mine drilling workers
Zhiming LIANG ; Bin XIAO ; Maosheng YAN ; Ting CHEN ; Bing YU ; Hansheng LIN ; Xiao ZHANG ; Hua YAN ; Peng WU ; Danying ZHANG ; Qingsong CHEN
China Occupational Medicine 2019;46(04):407-411
OBJECTIVE: To analyze the characteristic change of fingertip vibrotactile perception threshold(VPT) at two different frequencies among mine drilling workers. METHODS: A total of 48 mine drilling workers exposed to hand-transmitted vibration for at least 1.0 year were selected from mines in Hubei Province as the vibration exposure group by using the random number table method, and workers without hand-transmitted vibration exposure were selected as the control group. An HVLab vibrotactile perception meter was used to measure the fingertip VPT at 31.5 and 125.0 Hz in workers of these two groups. RESULTS: None of the workers in these two groups showed any clinical symptoms associated with occupational hand-arm vibration diseases. At 31.5 and 125.0 Hz, the fingertip VPT in the vibration exposure group was higher than that in the control group(P<0.01), the fingertip VPT of the little fingers was higher than that of the index fingers(P<0.01). At 125.0 Hz, the fingertip VPT of the fingers in dominant hands was higher than that in non-dominant hands in workers of the vibration exposure group(P<0.01). CONCLUSION: The mine drilling workers with hand-transmitted vibration exposure had higher fingertip VPT. There were differences of VPT in fingers and dominant hands.
10.Transumbilical endoscopic surgery in the diagnosis of ascites of unknown origin.
Yu LONG ; Anliu TANG ; Li TIAN ; Weiwei LUO ; Rui LIU ; Zhiming XIAO ; Xiaoyan WANG
Journal of Central South University(Medical Sciences) 2019;44(6):634-641
To explore whether transumbilical endoscopic surgery (TUES) can effectively and safely elucidate the causes of ascites of unknown origin.
Methods: A total of 23 consecutive patients with ascites of unknown origin who undertook TUES procedures in the Department of Gastroenterology of The Third Xiangya Hospital of Central South University between January 2014 and January 2016 were retrospectively investigated. Clinical manifestations, laboratory examinations and findings from radiological examinations and endoscopic investigations were noted before the procedure. Conditions of the abdominal cavity under endoscope, final diagnosis and outcome of patients were carefully recorded.
Results: TUES procedure was successfully performed in all 23 patients with an operation time of (58.2±13.9) min. Twenty-two patients were undertaken biopsy on the nodules or masses that found in the abdominal cavity. Definite diagnoses were established in the overwhelming majority of patients (22/23). The most common causes of ascites for the 23 cases was tuberculosis (8 cases), followed by peritoneal carcinomatosis (6 cases), and pseudomyxoma peritonei (5 cases). Operation-related complications, such as postoperative bleeding, perforation, peritonitis, intraabdominal chronic abscesses, were not observed, except one case showed a transient moderate fever in 24 hours after operation. No mortality related to TUES occurred. We concluded that TUES was a feasible, economic and minimally invasive approach to access the peritoneal cavity.
Conclusion: TUES combinated with biopsy can effectively elucidate the causes of ascites of unknown origin.
Ascites
;
Humans
;
Laparoscopy
;
Operative Time
;
Pseudomyxoma Peritonei
;
Retrospective Studies

Result Analysis
Print
Save
E-mail