1.Involvement of miR-15b-5p in depression-like behavior in Parkinson's disease by down-regulating BDNF/TrkB/PSD95 expressions
Hanjiang LUO ; Chengli WU ; Jinyong ZHU ; Min CHEN
Chinese Journal of Neuromedicine 2024;23(7):649-658
Objective:To explore the mechanism of miR-15b-5p involving in depression-like behavior in Parkinson's disease (PD).Methods:(1) Eighteen C57BL/6N mice were randomly divided into PD group, intervention group and control group ( n=6). PD models in PD group were established by stereotaxically injecting 0.25 mg/kg rotenone into the right striatum; mice in the intervention group were injected with 0.25 mg/kg rotenone and miR-15b-5p inhibitor lentivirus, while mice in the control group were injected with equal volume of PBS. Four weeks after that, open field test and rotarod test were performed to evaluate the motor ability, and sucrose preference and forced swimming tests were performed to evaluate the depression-like behaviors. And then, proteins and miRNAs in the substantia nigra were extracted; real-time fluorescent quantitative reverse transcription PCR (qRT-PCR) was used to detect the miR-15b-5p expression,and Western blotting and immunofluorescent staining were used to detect the tyrosine hydroxylase (TH), brain-derived neurotrophic factor (BDNF), tropomyosin receptor kinase B (TrkB), and postsynaptic density protein 95 (PSD95) protein expressions. (2) The 100 nmol/L miR-15b-5p mimic/inhibitor and their negative control sequences were transfected into SH-SY5Y cells on 6-well plates (named miR-15b-5p mimic group, miR-15b-5p mimic control group, miR-15b-5p inhibitor group and miR-15b-5p inhibitor control group, respectively); 48 h after that, BDNF, TrkB and PSD95 protein expressions were detected by Western blotting and miR-15b-5p expression by qRT-PCR. (3) The 100 ng BDNF 3'-UTR wild-type or mutant luciferase reporter vector plasmids and 100 nmol/L miR-15b-5p mimic/inhibitor or their negative control sequences were co-transfected into SH-SY5Y cells on 24-well plates, and luciferase reporter activity assay was performed 48 h after co-transfection to detect the luciferase activity. Results:(1) Compared with the control group, the PD group had significantly reduced movement speed, shortened rotarod drop latency, decreased percentage of sucrose preference, and prolonged immobility time ( P<0.05); compared with the PD group, the intervention group had significantly increased movement speed, prolonged rotarod drop latency, increased percentage of sucrose preference, and shortened immobility time ( P<0.05). (2) Compared with the miR-15b-5p mimic control group, the miR-15b-5p mimic group had significantly increased miR-15b-5p expression, and decreased BDNF, TrkB and PSD95 protein expressions (100.00±5.75 vs. 66.79±5.90; 100.00±5.95 vs. 84.46±5.77; 100.00±7.02 vs. 80.43±3.25, P<0.05). Compared with the miR-15b-5p inhibitor control group, the miR-15b-5p inhibitor group had significantly decreased miR-15b-5p expression, and increased BDNF, TrkB and PSD95 expressions (100.00±6.81 vs. 119.90±5.66; 100.00±2.88 vs. 110.10±4.15; 100.00±2.19 vs. 124.60±11.69, P<0.05). Compared with the control group, PD group had significantly increased miR-15b-5p expression, and significantly decreased TH, BDNF, TrkB and PSD95 expressions and BDNF fluorescent intensity (100.00±9.20 vs. 63.60±12.80; 100.00±9.88 vs. 71.95±10.00; 100.00±5.16 vs. 70.37±8.43; 100.00±7.01 vs. 68.12±10.22; 100.00±12.99 vs. 48.23±12.58) in the substantia nigra ( P<0.05); compared with the PD group, the intervention group had significantly lower miR-15b-5p expression and increased TH, BDNF, TrkB and PSD95 expressions and BDNF fluorescent intensity (63.60±12.80 vs. 90.69±9.84; 71.95±10.00 vs. 93.31±4.50; 70.37±8.43 vs. 88.11±4.10; 68.12±10.22 vs. 89.59±5.93; 48.23±12.58 vs. 83.65±10.52) in the substantia nigra ( P<0.05). (3) Compared with the BDNF 3'-UTR wild-type+miR-15b-5p mimic control group, the BDNF 3'-UTR wild-type+miR-15b-5p mimic group had significantly decreased luciferase activity (100.00±5.07 vs. 90.59±1.75, P<0.05); compared with the BDNF 3'-UTR wild-type+miR-15b-5p inhibitor control group, the BDNF 3'-UTR wild-type+miR-15b-5p inhibitor group had significantly increased luciferase activity (100.00±5.08 vs. 152.20±31.87, P<0.05). Conclusion:MiR-15b-5p is involved in depression-like behavior in PD by down-regulating the BDNF/TrkB/PSD95 expressions.
2.The analysis of unconventional lymph node metastasis in tongue squamous cell carcinoma
Kun WU ; Bowen DAI ; Hanjiang WU
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2024;59(3):227-232
Objective:To analyze the characteristics of cervical lymph node metastasis in tongue squamous cell carcinoma (TSCC).Methods:A retrospective study was conducted. A total of 329 patients with TSCC who underwent en bloc resection of primary tumor and neck dissection in the Second Xiangya Hospital of Central South University from June 2010 to March 2018 were included. There were 283 males and 46 females, aged from 26 to 80 years. All patients underwent the modified neck dissection. The main difference between the modified neck dissection and the traditional neck dissection lay in the managements of unconventional lymph nodes. The lymphatic adipose tissues adjacent to the superior thyroid artery, the base of facial artery and the branches of external carotid artery were thoroughly dissected. The primary tumor as well as lingual artery, tissues along the lingual artery and lymph nodes in the mouth floor were resected. χ 2 test was used for comparison of count data, and linear regression model was used for multivariate analysis. Results:Cervical lymph node metastases were found in 136 patients (41.3%). Among 142 patients (T1-2cN0) with supraomohyoid neck dissection, 22 patients had pathologically occult lymph node metastases (15.5%), with a 5-year overall survival rate of 90.2%, which was similar to the 5-year overall survival rate of 92.1% in 120 patients without lymph node metastasis (χ 2=0.156, P=0.693). Multivariate linear regression analysis showed that T stage, clinical stage and unconventional lymph node metastasis were important factors for cervical lymph node metastasis in tongue cancer patients ( P<0.05). Unconventional lymph node metastases occurred in 30 patients (9.1%), including the metastases of lymph nodes in the floor of mouth (3.0%), the lingual artery (2.4%), the base of the external maxillary artery (2.1%), the superior thyroid artery (0.9%), and the external carotid artery (0.6%). There were significant differences in the unconventional lymph node metastasis rates between patients with negative and positive conventional lymph node metastases [4.9%(10/203) vs. 15.9%(20/126), χ 2=11.242, P=0.001] and also between patients with depth of invasion ≤5 mm, 5 mm
3.A new surgical strategy for T4 stage posterior buccal carcinoma: anatomical unit resection
Kun WU ; Yuanyuan MAO ; Hanjiang WU
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2024;59(9):928-933
Objective:To investigate the effect of anatomical unit resection on the overall survival rate of patients with T4 stage posterior buccal carcinoma.Methods:We conducted a retrospective study on 79 patients with T4 stage posterior buccal squamous cell carcinoma underwent radical surgery for buccal mucosa cancer and reconstruction with free anterolateral thigh flap in the Department of Oral and Maxillofacial Surgery, the Second Xiangya Hospital, Central South University from March 2014 to December 2019. The 74 patients were males and the 5 patients were females, aged from 28 to 72 years old. The 40 patients in control group underwent traditional extended resection (1.5-2.0 cm safe margin), and 39 patients in the experimental group underwent anatomical unit resection. The clinicopathological parameters of patients were collected and followed up regularly. The overall survival rate was assessed by Log-rank and Cox proportional risk regression model. Multivariate analysis was performed by linear regression model.Results:There were significant differences between experimental group and control group in overall survival rates (61.53% vs 27.50%, χ2=6.624, P=0.010) and local disease control rates (74.36% vs 27.50%, χ2=17.350, P<0.001). Multiple linear regression analysis showed that local disease control rate was significantly correlated with anatomical unit resection ( t=3.880, P<0.001) and lymph node metastasis ( t=2.619, P=0.011), and also Cox proportional hazards regression model analysis identified that overall survival rate was significantly correlated with anatomical unit resection ( Z=2.421, P=0.016) and lymph node metastasis ( Z=2.793, P=0.005). Conclusion:For posterior buccal carcinoma with multiple anatomical units involved, anatomical unit resection can significantly reduce local recurrence and improve overall survival rate of patients.
4.Novel discovery of schisandrin A regulating the interplay of autophagy and apoptosis in oligoasthenospermia by targeting SCF/c-kit and TRPV1 via biosensors.
Lijuan MA ; Boyi LI ; Jinchen MA ; Chunyuan WU ; Nan LI ; Kailin ZHOU ; Yun YAN ; Mingshuang LI ; Xiaoyan HU ; Hao YAN ; Qi WANG ; Yanfei ZHENG ; Zhisheng WU
Acta Pharmaceutica Sinica B 2023;13(6):2765-2777
Oligoasthenospermia is the primary cause of infertility. However, there are still enormous challenges in the screening of critical candidates and targets of oligoasthenospermia owing to its complex mechanism. In this study, stem cell factor (SCF), c-kit, and transient receptor potential vanilloid 1 (TRPV1) biosensors were successfully established and applied to studying apoptosis and autophagy mechanisms. Interestingly, the detection limit reached 2.787 × 10-15 g/L, and the quantitative limit reached 1.0 × 10-13 g/L. Furthermore, biosensors were used to investigate the interplay between autophagy and apoptosis. Schisandrin A is an excellent candidate to form a system with c-kit similar to SCF/c-kit with a detection constant (KD) of 5.701 × 10-11 mol/L, whereas it had no affinity for SCF. In addition, it also inhibited autophagy in oligoasthenospermia through antagonizing TRPV1 with a KD of up to 4.181 × 10-10 mol/L. In addition, in vivo and in vitro experiments were highly consistent with the biosensor. In summary, high-potency schisandrin A and two potential targets were identified, through which schisandrin A could reverse the apoptosis caused by excessive autophagy during oligoasthenospermia. Our study provides promising insights into the discovery of effective compounds and potential targets via a well-established in vitro-in vivo strategy.
5.Critical quality attribute assessment of big brand traditional Chinese medicine: quality control method of Huangjing Zanyu Capsules based on chemical properties of characteristic components.
Jin-Yuan ZHU ; Jing-Qi ZENG ; Qian-Qian LI ; Li-Juan MA ; Ying LU ; Han ZHANG ; Yan-Fei ZHENG ; Chun-Yuan WU ; Zhi-Sheng WU
China Journal of Chinese Materia Medica 2021;46(7):1629-1635
The chemical properties of characteristic components are significant to the manufacturing quality control of big brand traditional Chinese medicine. In this study, the Huangjing Zanyu Capsules were used as the research carrier to determine the content of five characteristic components including icraiin, emodin, schisandrin A, 2,3,5,4'-tetrahydroxystilbene-2-O-β-D-glucoside, and osthole simultaneously by high-performance liquid chromatography(HPLC). The results showed that the chemical properties of five cha-racteristic components had a good linear relationship(r>0.999 9) within the quantitative range; the relative standard deviations(RSD) was 0.11%-2.0% and 0.25%-2.8% respectively for intra-day and inter-day precision; the RSD of repeatability was 1.8%-2.6%; the RSD of stability within 48 hours was 0.19%-2.8%, and the average recovery rate was 95.52%-100.1%, all meeting the requirements of pharmaceutical quantitative analysis. Additionally, the interval estimation method was used to directly reflect the distribution of samples with abnormal chemical properties of characteristic components, and the results showed ten samples were detected beyound the 95% control line of confidence level. Multivariate statistical process control(MSPC) method was used to monitor the abnormal samples of Huangjing Zanyu Capsules collectively, and the results showed that two samples were beyond the 95% control line of Hotelling's T~2 and three samples beyond the 95% control line of squared prediction error(SPE), indicating consistent quality control of Huangjing Zanyu Capsules. In conclusion, the proposed method is not only accurate and efficient but also a compensation for the traditional single-component quality control method, providing a scientific basis for the quality control in manufacturing process of Huangjing Zanyu Capsules. Furthermore, it could also serve as a reference method for the quality control in manufacturing big brand traditional Chinese medicine.
Capsules
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Chromatography, High Pressure Liquid
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Drugs, Chinese Herbal
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Medicine, Chinese Traditional
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Quality Control
6.Effect and cost-effectiveness of three commonly used molluscicides in largescale field application
Hong-Chu WU ; Yu-Cai MA ; Zheng-Qiu ZHANG ; Kai TANG ; Guang-Ling DU ; Jin ZHANG ; Jian WANG ; Dao-Jian ZHU ; Yin-Ping ZUO
Chinese Journal of Schistosomiasis Control 2018;30(6):619-624
Objective To evaluate the effect and cost-effectiveness of three commonly used molluscicides, 4% "Luo-wei" (tea-seed distilled saponins, TDS), 50% niclosamide ethanolamine salt wettable powder (NESWP), and 26% metaldehyde and niclosamide suspension concentrate (MNSC) in large-scale field application, so as to provide the references for formulating the strategy of snail control. Methods The field test and parallel comparison were implemented. A marshland with Oncomelania hupensis snails of the Yangtze River was divided into 4 parts (10 hm2) for the research, and three of them were experimental areas while the last one was a blank control area. The experimental areas were sprayed with 4% "Luo-wei", 50% NESWP and 26% MNSC respectively for 3 times and the interval was 1 week. Seven days after each spraying the effect of snail control was investigated, and the costs of molluscicides, labor, transportation, fuel consumption and mechanical loss were recorded. The cost of each molluscicide, snail mortality, snail density, and the cost of increasing 1% of snail mortality per 100 m2 were analyzed and compared. Results After the first, second and third spraying, the corrected snail mortality rates were 67.34%, 76.55% and 84.60% respectively in the 4% "Luo-wei" group; the corrected snail mortality rates were 64.71%, 75.17% and 83.89% respectively in the 50% NESWP group; the corrected snail mortality rates were 66.55%, 76.27% and 86.67% respectively in the 26% MNSC group. There was no significant difference among the 3 groups in the snail mortality at the same spraying time (χ2 = 1.590, 0.571, 3.238, all P > 0.05) . In addition, along with the increase of the spraying times, the snail mortality of each group was increased significantly compared to that of the control group (χ2 = 79.333, 94.718, 117.020, all P < 0.01) . After the first, second and third spraying, the reduction rates of snail density were 69.82%–86.60% in the 4% "Luo-wei" group, 68.66%–86.55% in the 50% NESWP group, and 71.89%–88.87% in the 26% MNSC group respectively. The decreasing amplitude of the snail density was more than 85% in all the experimental areas after 3 rounds of spraying molluscicide. The snail control costs per 100 hm2 were 19.57, 11.97 Yuan and 10.47 Yuan in the 4% "Luo-wei" group, 50% NESWP group, and 26% MNSC group respectively. After the first, second and third spraying, the costs of increasing 1% of snail mortality per 100 m2 were 0.30, 2.08 Yuan and 2.38 Yuan in the 4% "Luo-wei" group, 0.20, 1.10 Yuan and 1.32 Yuan in the 50% NESWP group, and 0.17, 1.04 Yuan and 0.97 Yuan in the 26% MNSC group respectively, and the cost-effectiveness was the highest at the first spraying in all the three groups. Conclusions The effects of the three molluscicides for snail control are similar, but the efficacy of snail control is reduced as the spraying time increases.
7.Research progresses in occult lymph node metastasis of oral squamous cancer
Journal of Prevention and Treatment for Stomatological Diseases 2018;26(1):61-65
Oral cancer is a common malignant tumor in head and neck cancers, and the metastasis of cervical lymph nodes directly affects the prognosis of patients with oral cancer. Exploration and study on the detection of occult lymph nodes in patients with negative cervical lymph nodes is of great importance to improve the prognosis of patients with early oral cancer. This article reviews some progresses of occult lymph node metastasis of oral cancer in ultrasound guided fine-needle aspiration cytology (US-FNAC)、 positron emission tomography and single-photon emission computed tomography/computed tomography (SPECT/CT) fusion technology, sentinel lymph node biopsy technique (SLNB) and molecular biological detection、 nanotechnology and quantum dot technology.
8.How to deal with the vessel of free flap: A retrospective study of 264 cases of anterolateral thigh flap
Bo LI ; Zhenhu REN ; Kai WANG ; Mei CHEN ; Hanjiang WU
Chinese Journal of Microsurgery 2016;39(3):237-240
Objective To compare the reliability and effectiveness of anterolateral thigh flap with double vein anastomosis or one vein anastomosis for reconstruction of head and neck defects.Methods Two hundred and sixty four cases of anterolateral thigh flap transfers for head and neck reconstruction from January,2013 to September,2013 in the Second Xiangya Hospital of Central South University were reviewed.260 patients were randomly divided into 2 groups.In the experimental group,there were 138 patients 140 cases of anterolateral thigh flap with one vein one artery anastomosis.In the control group,there were 122 patients 124 cases of anterolateral thigh flap with double vein one artery anastomosis.Results Among 264 anterolateral thigh flaps,the overall success rate of free flap was 98.1% (259/264),5 free flaps were lost.In the experimental group,there were 6 free flaps occurred venous thrombosis,two of them were lost.In the control group,there were 5 cases occurred venous thrombosis,three of them were lost.No arterial thrombosis occurred in both groups.The time of micromanipulation was 18 to 101 min,with the average of 47 min in the experimental group.In the control group,the time was 45 to 133 min,with the average of 71 min.(P =0.0003).Conclusion Anterolateral thigh flap with one vein one artery anastomosis for head and neck reconstruction did not affect the survival rate but it can absolutely reduce the operation time.
9.Application of 1,212 anterolateral thigh myocutaneous flaps in the repair of oral and maxillofacial defects.
Zhen-hu REN ; Hanjiang WU ; Hongyu TAN ; Kai WANG ; Zhaojian GONG ; Sheng ZHANG ; Jinbing LIU ; Zhaofu ZHU
West China Journal of Stomatology 2015;33(3):281-285
OBJECTIVETo examine the benefits of anterolateral thigh myocutaneous flaps in reconstruction of oral and maxillofacial defects.
METHODSPatients were recruited from February 2002 to June 2013 in the Department of Oral and Maxillofacial Surgery of Central South University. All patients (1,185 patients, 1,212 transferred flaps) underwent reconstructive surgery employing anterolateral thigh myocutaneous flaps. Basic information for all patients including defect side, flap size and type, recipient vessel processing method, donor complications, and postoperative quality of life were recorded and statistically analyzed.
RESULTSAmong the 1 212 transferred flaps, 1 176 survived and 36 showed necrosis, for a survival rate of about 97.0%. No cases presented with local serious complications, and 90% of patients achieved good functional recovery and aesthetically acceptable results after reconstruction of oral and maxillofacial defects at various locations using anterolateral thigh myocutaneous flaps. The time for anastomosis of one vein was significantly less than that for two veins (P=-0.000 3), which indicated one vein anastomosis could significantly reduce the operating time. The incidence of venous crisis, the survival rate after treatment, and the rate of venous crisis resulting in flap necrosis were comparable between the groups (P>0.05).
CONCLUSIONAnterolateral thigh myocutaneous flaps can be easily obtained and provide a good amount of muscle for filling dead space and fascia lata. These flaps can meet the various requirements of oral and maxillofacial defects. Therefore, the anterolateral thigh myocutaneous free flaps are more suitable for oral and maxillofacial defects than other flaps.
Free Tissue Flaps ; Humans ; Maxillofacial Abnormalities ; surgery ; Myocutaneous Flap ; Necrosis ; Quality of Life ; Reconstructive Surgical Procedures ; methods ; statistics & numerical data ; Surgery, Oral ; Thigh ; Wound Healing
10.Application of 909 anterolateral thigh myocutaneous flaps in the reconstruction of oral and maxillofacial defects
Bo LI ; Zhenhu REN ; Kai WANG ; Mei CHEN ; Hanjiang WU
Chinese Journal of Stomatology 2015;50(3):169-172
Objective To summarize the application of 909 anterolateral thigh myocutaneous flaps in the repair of oral and maxillofacial defects and to examine their benefits in maxillofacial reconstruction of these defects.Methods Patients were recruited from January 2004 to December 2012 in the Department of Oral and Maxillofacial Surgery of the Second Xiangya Hospital of Central South University.All patients underwent reconstructive surgery with anterolateral thigh myocutaneous flaps,and patient age ranged from 19 to 81 years with a mean of 51.2 years.There were 761 flaps showing single lobe and 148 flaps showing a multi-island pedicle.The largest area among the single flaps was 28 cm× 12 cm,and the smallest was 4 cm× 2 cm.Results Among the 909 transferred flaps,882 survived and 27 showed necrosis,with a survival rate of about 97.0%.The common complications at flap donor site were poor wound healing 9.6%(87/909),localized paresthesia 61.0%(500/820),and altered quadriceps force 15.0%(123/820).No case was presented with local serious complications,and 90% of the patients achieved good functional recovery and aesthetically acceptable results after the reconstruction by anterolateral thigh myocutaneous flaps.Conclusions The anterolateral thigh myocutaneous free flaps are more suitable for oral and maxillofacial defects than other flaps and should be preferred.


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