1.Single-cell RNA Sequencing Reveals Thoracolumbar Vertebra Heterogeneity and Rib-genesis in Pigs.
Jianbo LI ; Ligang WANG ; Dawei YU ; Junfeng HAO ; Longchao ZHANG ; Adeniyi C ADEOLA ; Bingyu MAO ; Yun GAO ; Shifang WU ; Chunling ZHU ; Yongqing ZHANG ; Jilong REN ; Changgai MU ; David M IRWIN ; Lixian WANG ; Tang HAI ; Haibing XIE ; Yaping ZHANG
Genomics, Proteomics & Bioinformatics 2021;19(3):423-436
Development of thoracolumbar vertebra (TLV) and rib primordium (RP) is a common evolutionary feature across vertebrates, although whole-organism analysis of the expression dynamics of TLV- and RP-related genes has been lacking. Here, we investigated the single-cell transcriptome landscape of thoracic vertebra (TV), lumbar vertebra (LV), and RP cells from a pig embryo at 27 days post-fertilization (dpf) and identified six cell types with distinct gene expression signatures. In-depth dissection of the gene expression dynamics and RNA velocity revealed a coupled process of osteogenesis and angiogenesis during TLV and RP development. Further analysis of cell type-specific and strand-specific expression uncovered the extremely high level of HOXA10 3'-UTR sequence specific to osteoblasts of LV cells, which may function as anti-HOXA10-antisense by counteracting the HOXA10-antisense effect to determine TLV transition. Thus, this work provides a valuable resource for understanding embryonic osteogenesis and angiogenesis underlying vertebrate TLV and RP development at the cell type-specific resolution, which serves as a comprehensive view on the transcriptional profile of animal embryo development.
2.Effect of biglycan on neural apoptosis in mice with early brain injury after subarachnoid hemorrhage
Yuke XIE ; Kecheng GUO ; Jianhua PENG ; Jinwei PANG ; Jian ZHOU ; Long GU ; Lifang ZHANG ; Shigang YIN ; Ligang CHEN ; Yong JIANG
Chinese Journal of Neuromedicine 2020;19(2):138-145
Objective To investigate the effect ofbiglycan (BGN) on neural apoptosis in mice with early brain injury (EBI) after subarachnoid hemorrhage (SAH).Methods SAH models were induced by endovascular perforation in young male C57BL/6J mice.(1) Totally,48 mice were randomly divided into sham-operated group,SAH 6 h group,SAH 12 h group,SAH 24 h group,SAH 48 h group,and SAH 72 h group (n=8);the BGN protein and mRNA expressions were detected by Western blotting and real-time quantitative PCR (qRT-PCR).(2) Totally,16 mice were randomly divided into sham-operated group and SAH 48 h group (n=8);immunofluorescent double staining was conducted to explore the BGN expression in the neurons of brain tissues.(3) Totally,24 mice were randomly divided into sham-operated group,sham+control lentivirus group,and sham+BGN lentivirus group (n=8);BGN lentiviral vector and control lentivirus were administered intracerebroventricularly 7 d before sham-operation;qRT-PCR was performed to explore the BGN mRNA expression.(4) Totally,48 mice were randomly divided into sham-operated group,SAH+control lentivirus group,and SAH+BGN lentivirus group (n=16);BGN lentiviral vector and control lentivirus were administered intracerebroventricularly 7 d before SAH;neurological scores were detected by modified Garcia scale and beam balance tests;TUNEL was used to detect the neuronal apoptosis,and Western blotting was performed to explore the expressions of nuclear transcription factor kappa B (NF-κB) and phosphorylated-(p-) NF-κB.Results (1) Mice in the SAH 48 h group had the highest BGN protein and mRNA expressions,which showed statistical differences as compared with the sham-operated group (P<0.05).(2) A majority of BGN expressions were detected in the neurons 48 h after SAH.(3) The sham+BGN lentivirus group had statistically lower BGN mRNA expression than the sham+control lentivirus group (P<0.05).(4) As compared with those in the SAH+control lentivirus group,both scores of modified Garcia scale and beam balance tests were significantly higher in SAH+BGN lentivirus group (6.125±1.246 vs.13.000±1.309;1.125±1.126 vs.2.875±0.835),and neural apoptosis ratio and ratio of p-NF-κB/NF-κB were significantly lower in the SAH+BGN lentivirus group (51.950%±11.166% vs.31.938%±7.705%;1.161±0.156 vs.0.886±0.142,P<0.05).Conclusion Inhibition of BGN can effectively reduce neuronal apoptosis in mice with EBI after SAH,and attenuate neurological deficits.
3.Effect of long non-coding RNA F19 on secondary brain injury after traumatic brain injury in mice
Jianhua PENG ; Jinwei PANG ; Yue WU ; Yuke XIE ; Kecheng GUO ; Tianqi TU ; Qiancheng MU ; Yuyan LIAO ; Fang CAO ; Liang LIU ; Ligang CHEN ; Xiaochuan SUN ; Yong JIANG
Chinese Journal of Trauma 2019;35(3):267-273
Objective To investigate the effect of long non-coding RNA F19 (lncRNA F19) on secondary brain injury following traumatic brain injury (TBI) in mice. Methods (1) A total of 96 C57BL/6J male wild-type mice were divided into sham group, sham+control lentivirus group, sham+F19 lentivirus group, TBI group, TBI+control lentivirus group and TBI+F19 lentivirus group according to the random number table. Each group consisted of two subgroups of 1 day and 3 days after TBI, with eight mice per subgroup. The expression and silence efficiency of lncRNA F19 were detected. ( 2 ) A total of 96 C57BL/6J male wild-type mice were divided into sham group, TBI+control lentivirus group and TBI + F19 lentivirus group according to the random number table. Each group consisted of two subgroups of 1 day and 3 days after TBI, with 16 mice per subgroup. The effect of lncRNA F19 on neuronal apoptosis after TBI was recorded. The mice TBI model was established using the controlled cortical damage method (CCI). The lncRNA F19 lentivirus or control lentivirus were administrated by intracerebroventricular injection 5 days before injury. The expressions of lncRNA F19 ( 2 -ΔΔct ) were detected by real-time quantitative PCR ( qRT-PCR ) at 1 day and 3 days after injury. The Toll-like receptor 4 (TLR4), B lymphocyte tumor-2 (Bcl-2) and Bcl-2 related protein (Bax) expressions were detected by Western blot. The TUNEL was used to detect apoptosis around the traumatic lesions. Results From the first day after injury, both in the sham operation and TBI groups, the control lentivirus had no effect on the level of lncRAN F19 (P >0. 05). One day after injury, compared with sham +control lentivirus group, the levels of lncRNA F19 in sham + F19 lentivirus group were significantly decreased (0. 07 ± 0. 07:0. 93 ± 0. 17);compared with TBI+control lentivirus group, levels of lncRNA F19 in TBI+F19 lentivirus group were significantly decreased (2. 91 ± 1. 18:0. 52 ± 0. 32) (P<0. 05). There were significantly lower protein levels of TLR4 (0. 51 ± 0. 13:0. 66 ± 0. 15), Bax (0. 45 ± 0. 06:0. 67 ± 0. 16), lower TUNEL-positive neurons ratio [(23. 55 ± 6. 85)% : (31. 58 ± 7. 52)%], but higher protein levels of Bcl-2 (0. 76 ± 0. 16:0. 47 ± 0. 12) in TBI+F19 lentivirus group compared with the TBI+ control lentivirus group (P <0.05). Three days after injury, compared with sham + control lentivirus group, levels of lncRNA F19 in sham+F19 lentivirus group were significantly decreased (0. 11 ± 0. 09:0. 96 ± 0. 09); compared with TBI+control lentivirus group, levels of lncRNA F19 in TBI+F19 lentivirus group were significantly decreased (0. 54 ± 0. 24:3. 39 ± 0. 90) (P <0. 05). There were significantly lower protein levels of TLR4 (0. 60 ± 0. 20):(0. 85 ± 0. 09)], lower Bax (0. 60 ± 0. 12:0. 88 ±0. 21), lower TUNEL-positive neurons ratio [(29. 10 ± 7. 37)% :(39. 22 ± 10. 64)%], but higher protein levels of Bcl-2 (0. 66 ± 0. 12:0. 35 ± 0. 16) in TBI+F19 lentivirus group compared with the TBI+control lentivirus group (P<0. 05). Conclusion Inhibition of lncRNA F19 can significantly reduce the TLR4-induced neuronal apoptosis in cortex after TBI in mice and alleviate reduce the secondary brain injury.
4.Effect of acupoint catgut embedding, fire needle, auricular acupuncture on female post-adolescent acne and serum sex hormone.
Wen LU ; Ligang ZHU ; Qianmo TIAN ; Xinyu BAI ; Weiwei KUANG ; Zhongnv FANG ; Hong REN ; Renqiong CHEN ; Chuntao XIE
Chinese Acupuncture & Moxibustion 2018;38(8):833-838
OBJECTIVETo compare the difference of serum sex hormone between female patients with post-adolescent acne and healthy women, and to explore the efficacy and action mechanism of acupoint catgut embedding, fire needle, auricular acupuncture on skin lesion in female patients of post-adolescent acne.
METHODSA total of 107 female patients of post-adolescent acne were divided into an integrated acupuncture group (54 cases, 4 cases were excluded) and a medication group (53 cases, 5 cases were excluded). The patients in the integrated acupuncture group were treated with comprehensive treatment of acupoint catgut embedding, fire needle, auricular acupuncture; the acupoint catgut embedding was applied at Dazhui (GV 14), Yintang (GV 29), Yangbai (GB 14) through Yuyao (EX-HN 4) and other acupoints based on syndrome differentiation; the fire needle was applied at skin lesion; the auricular acupuncture was applied at erjian (HX), e (AT), kou (CO), etc. The patients in the medication group were treated with oral administration of tanshinone capsules (4 capsules each time, 3 times a day) and external use of adapalene gel (one treatment per day at night). Patients in the two groups were treated for 8 weeks. The skin lesion of acne was evaluated before treatment as well as 4 weeks and 8 weeks after treatment in the two groups; the serum levels of testosterone (T) and estradiol (E) were tested 24 hours before menstruation in the integrated acupuncture group (50 cases) and healthy control group (46 cases), and the change of serum sex hormone after treatment was observed in 21 patients with sex hormone disorder in the integrated acupuncture group.
RESULTSBefore treatment, the level of E in the integrated acupuncture group was significantly lower than that in the healthy control group (<0.01), but T/E in the integrated acupuncture group was significantly higher than that in the healthy control group (<0.01). After treatment, the level of E was significantly increased (<0.01) and T/E was reduced (<0.01) in the 21 patients with sex hormone disorder in the integrated acupuncture group. The skin lesion scale of acne was significantly reduced in the two groups after 4-week and 8-week treatment (all <0.01); the difference between the two groups was not significant after 4-week treatment (>0.05); the skin lesion scale of acne in the integrated acupuncture group was significantly lower than that in the medication group after 8-week treatment (<0.01). The efficacy between the two groups was not significant after 4-week the treatment (>0.05); after 8-week treatment, the cured and effective rate was 66.0% (33/50) in the integrated acupuncture group, which was superior to 45.8% (22/48) in the medication group (<0.05).
CONCLUSIONCompared with healthy women, the level of serum sex hormone of E is reduced in the female patients of post-adolescent acne, resulting in relative increased level of T; the acupoint catgut embedding, fire needle, auricular acupuncture have better efficacy than medication for post-adolescent acne, which have regulation effects on sex hormone disorder.
5.Establishment of chemiluminescent enzyme immunoassay for detecting antibodies against foot-and-mouth disease virus serotype O in swine.
Chen CUI ; Ligang HUANG ; Jing LI ; Xingqi ZOU ; Yuanyuan ZHU ; Lei XIE ; Qizu ZHAO ; Limin YANG ; Wenjun LIU
Chinese Journal of Biotechnology 2016;32(11):1519-1530
Recombinant structural protein VP1 of foot-and-mouth disease virus serotype O was expressed in Escherichia coli and then purified using Nickel affinity chromatography. A chemiluminescent enzyme immunoassay (CLEIA) method was established using the purified recombinant protein as coating antigen to detect antibody of foot-and-mouth disease virus serotype O in swine. The specificity of VP1-CLEIA method is 100%. The coefficients of variation in the plate and between plates are 1.10%-6.70% and 0.66%-4.80%, respectively. Comparing with the commercial indirect ELISA kit or liquid phase block ELISA kit, the calculated coincidence rate is 93.50% or 94.00%. The high specificity and stability suggested this detection method can be used to monitor the antibody level of foot-and-mouth disease virus serotype O in swine.
6.Association study on lifestyle and internet addiction in adolescents
Hua DONG ; Ligang WANG ; Dongjie XIE ; Wenbin GAO ; Xinying SUN
Chinese Journal of Behavioral Medicine and Brain Science 2015;24(7):633-636
Objective To investigate the lifestyles of internet addiction disorders and explore the relationship between lifestyles and internet addiction among Chinese adolescents.Methods In this cross-sectional stratified cluster sampling study,Demographic Information Questionnaire,Chinese Adolescent Lifestyles Scale (CALS)and Young's Internet Addiction Test were applied.Results (1) Adolescents with internet addiction were 275,accounting for 11.20%.Boys(15.45%) addicted the internet more than girls(7.48%).The prevalence rate of IAD was higher in students from urban(13.55%) than that in students from rural area(9.67%).The ratio of IAD was the highest in junior school students (14.01%),the second in university students (13.84%),the third in senior high school students(9.66%),and the lowest in primary school students(7.67%) (P<0.01).(2) The internet addicts scored significantly lower on sleep,study habits,physical exercise,interpersonal relationship and coping style,compared to the normal group(P<0.01).(3) Sleep status,study habits,physical exercise,interpersonal relationship and coping style were negatively correlated with internet addiction (r=-0.193,r=-0.319,r=-0.175,r=-0.225,r=-0.105,P<0.01).(4)Multiple logistic regression analysis indicated learning habits and interpersonal relationship had negative predictive effects on internet addiction; whereas,the coping style showed a weak positive prediction effect on Internet addiction.Conclusion Internet addict teenagers have poor lifestyles which should be suggested to start the good behavior habit and insist on a healthy lifestyle to prevent the internet addiction disorder.
7.Content Determination of Chlorogenic Acid in Yinhuang Qinghuo Capsule by HPLC
Ligang XIE ; Hejun CHEN ; Shixue HE ; Fang HE
China Pharmacy 2015;26(36):5147-5148
OBJECTIVE:To establish a method to determine the content of chlorogenic acid in Yinhuang qinghuo capsule. METHODS:HPLC was performed on the column of Acclaim120? C18 with mobile phase of acetonitrile-0.4% phosphoric acid(12:88,V/V)at flow rate of 1 ml/min,the detection wavelength was 327 nm,column temperature was 25 ℃,and volume injection was 10 μl. RESULTS:The linear range of chlorogenic acid was 56-224 μg;RSDs of precision,stability and reproduciblity tests were lower than 3.0%,recovery was 96.91%-104.76%(RSD=1.46%,n=9). CONCLUSIONS:The method is simple and reproducible, and can be used for the quality control of Yinhuang qinghuo capsule.
8.Development of adolescent executive function scale
Chunhui HUANG ; Yicheng TANG ; Ligang WANG ; Dongjie XIE ; Chunlei FAN ; Wenbin GAO
Chinese Journal of Behavioral Medicine and Brain Science 2014;23(5):463-465
Objective To develop a scale of the executive functional status for adolescents.Methods Regarding to the structure and the conception of executive function and the results of interviews to several adolescents,a preliminary inventory including 37 items was developed.The investigation sample consisted of 1722 students from 2 provinces in China by stratified sampling,and the the internal consistency,conduct validity and criterion validity were assessed by exploratory factor analysis.Results The revised scale including 21 items was consisted of three factors:inhibition control,cognitive flexibility,working memory,which explained 45.39% of the total variance.The factor loading ranged from 0.499 to 0.727.The Cronbach alpha ranged from 0.786-0.897 for the scale and its three factors.Factor structure fitted actual data and its fit indices were x2=530.44,df=186,RESMA =0.053,GFI =0.93,AGFI =0.91,NFI =0.96,CFI =0.97.Conclusion It shows that the reliability and validity of the scale meet the standard of the psychometrics,and it can be suitable for evaluating the functional status of execution of Chinese adolescent.
9.Impacting factors on the professional identity of railway young workers and the relationship with the professional identity and subjective well-being
Fuhua KUANG ; Dongjie XIE ; Ligang WANG ; Xiaolan FAN ; Wenbin GAO
Chinese Journal of Behavioral Medicine and Brain Science 2014;23(3):254-257
Objective To investigate the impacting factors on the professional identity of railway young workers and the relationship of the professional identity and subjective well-being.Methods The sample consisting of 5 762 young workers was selected from railway enterprises by stratified and random sampling according to the proportion.Occupational professional identity Scale and Index of Well-being,Index of General Affect Scale were used.The affect of the variable,such as sex,age,working years,educational background and income on professional identity were studied.Moreover,the relationship between professional identity and subjective well-being was reported.Results (1) Professional identity ascended witb the increase of ages,and there was significance between three groups(F=6.667,P<0.05).(2)Professional identity ascended with the increase of length of service (F=5.147,P<0.01),but it dropped at the point of 9-12 years.(3) The professional identity of workers with bachelor degree was higher than that with junior college education background.and there was significance between three groups (F=3.538,P<0.05).(4)The workers who had more than 3 500 RMB monthly salary was regarded on the top in professional identity,while those whose monthly salary were under 2 500 RMB was in the lowest level (F=42.275,P<0.01).(5)The result of linear regression indicated that the degree of professional identity can forecast the degree of subjective well-being(R =0.534,P<0.01).Conclusion The professional identity of railway young works was affected by their ages,the length of service,the educational degree and salary,The professional identity can forecast subjective well-being.
10.The dual-systems of self-control scale for children: development, reliability and validity
Ligang WANG ; Dongjie XIE ; Chunlei FAN ; Wenbin GAO
Chinese Journal of Behavioral Medicine and Brain Science 2014;23(4):362-365
Objective To develop and test the reliability and validity of the dual-systems of self-control scale for children (DSSCSC).Methods The scale was developed based on previous studies and the theoretical framework of the dual system.Totally 1493 families from 5 provinces were recruited.Participants were asked to complete the DSSCSC.Item analysis was conducted to identify valid items.Then the construct validity of the scale was tested through exploratory and confirmatory factor analysis(EFA & CFA).Finally the reliability and criterion validity of the scale was examined.Results The scale was consisted of 23 items with 5 factors-planning,impulsivity,distractibility,persistence,and poor delay of gratification.They explained 50.46% of the total variance.Besides,the CFA resuhs demonstrated a satisfactory construct validity of the scale (RMSEA =0.044,NFI =0.95,NNFI=0.97,CFI=0.97).The scores of DSSCSC and its subscales were significantly correlated with the TSCRS scores.The Cronbach' s α coefficients of the total scale was 0.855,and 5 subscales were 0.813,0.803,0.582,0.608,and 0.620 respectively.Conclusion The DSSCSC with 5 factors has a good reliability and validity,demonstrating it a valid tool to measure the children' s self-control in China.

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