1.Effect of MSI2 on proliferation and apoptosis imbalance of KGN cells induced by DHT
Youwen Du ; Haitao Shi ; Shuailong Han ; Shumin Zhang ; Chaoqi Guan ; Tianjuan Wang ; Linxin Pan
Acta Universitatis Medicinalis Anhui 2023;58(3):464-470
Objective:
To study the effect of Musashi-2 ( MSI2 ) overexpression on the imbalance of proliferation and apoptosis of human ovarian granulosa cell line (KGN) induced by dihydrotestosterone (DHT) .
Methods:
The gene expression profiles of human ovarian granulosa cells ( GCs) in primary culture were statistically analyzed to screen the differentially expressed genes.pcDNA3. 1-MSI2 eukaryotic expression plasmid was constructed and transiently transfected into the KGN cells,and the overexpression effect of MSI2 was detected by Quantitative Real-time PCR (RT-qPCR) and Western blot.After overexpressing MSI2 in DHT induced KGN cells,MTT colorimetry and Edu staining were used to detect the proliferation of cells in each group,and flow cytometry ( FCM) was further used to detect the apoptosis of cells in each group.
Results:
The mRNA expression level of MSI2 gradually decreased during the primary culture of human ovarian GCs.And pcDNA3. 1-MSI2 was successfully constructed and transfected into KGN cells to improve the mRNA and protein expression levels of MSI2.Then MTT,EdU and FCM results showed that compared with the blank group,DHT induction could significantly reduce the proliferation rate and increase the apoptosis rate of KGN cells (P <0. 05) .However,after MSI2 overexpression,the proliferation rate of KGN cells increased and the apoptosis rate decreased (P <0. 05) ,which were close to the blank group.
Conclusion
Overexpression of MSI2 can effectively alleviate the imbalance of proliferation and apoptosis of KGN cells induced by DHT,indicating that MSI2 plays an important role in GCs growth and follicle development.
2.Predictive value of visceral fat index and lipid accumulation product on metabolic associated fatty liver disease in the population without overweight/obesity
Hongyan WANG ; Yupeng LIU ; Hongmei FU ; Ruiling XU ; Chaoqi YAN
Chinese Journal of Health Management 2023;17(11):848-853
Objective:To investigate the value of visceral fat index (VAI) and lipid accumulation product (LAP) on predicting metabolic associated fatty liver disease (MAFLD) in the population without overweight/obesity.Methods:This study is a cross-sectional study. The physical examination data derived from International Physical Examination and Health Center of the Second Affiliated Hospital of Harbin Medical University from January to December 2021 were collected. According to the inclusion and exclusion criteria, a total of 4 304 subjects without overweight/obesity aged from 18-75 were included in this study. The subjects were divided into two groups with MAFLD or without MAFLD, according to the diagnostic criteria of MAFLD provided by The Asian Pacific Association for the Study of the liver clinical practice guidelines for the diagnosis and management of metabolic associated fatty liver disease. Comparison of the clinical parameters (blood pressure, lipid, glucose) and obesity measurement indexes (BMI, VAI, LAP) between the two groups was analyzed. All subjects were respectively divided into four groups according to BMI, VAI and LAP quartile, which were defined as A, B, C, D. The prevalence of MAFLD in the population without overweight/obesity in quartile area groups of different obesity measurement indexes was calculated. Spearman′s rank correlation was used to evaluate the correlation between BMI, LAP, VAI and MAFLD in the population without overweight/obesity, respectively. Meanwhile, receiver operating characteristics (ROC) curves were used to calculate area under the curve (AUC) and evaluate the accuracy of BMI, VAI and LAP on predicting for MAFLD in the population without overweight/obesity.Results:The prevalence of MAFLD in the population without overweight/obesity was 10.87%. In the population without overweight/obesity, the clinical data blood pressure, total cholesterol (TC), low density lipoprotein cholesterol (LDL-C), triglyceride (TG) and fasting plasma glucose (FPG) in the group with MAFLD were significantly higher than those of the group without MAFLD (131/80 vs 113/70 mmHg, 5.29 vs 4.65 mmol/L, 3.21 vs 2.75 mmol/L, 1.87 vs 0.89 mmol/L, 5.60 vs 4.95 mmol/L; P<0.001), but high density lipoprotein cholesterol (HDL-C) of the group with MAFLD was significantly lower than that of the group without MAFLD (1.19 vs 1.49 mmol/L; P<0.001). The obesity measurement indexes (BMI, VAI, LAP) in the group with MAFLD were significantly higher than those of the group without MAFLD (22.10 vs 20.70 kg/m 2, 2.64 vs 1.00, 36.27 vs 12.48; all P<0.001). In the population without overweight/obesity, the prevalence of MAFLD was increased with the increase of BMI, VAI and LAP quartile area, and there was a linear correlation between MAFLD and obesity measurement indexes above. Spearman′s rank correlation analysis showed that correlation coefficient between obesity measurement indexes and MAFLD in the population without overweight/obesity was respectively LAP (0.427)>VAI (0.406)>BMI (0.282). ROC curve analysis showed that in the population without overweight/obesity, LAP had the highest accuracy on predicting MAFLD, with the AUC value of 0.896 (0.886-0.905), the optional cut-off value was 20.75, sensitivity and specificity was 85.9% and 79.0%, respectively. VAI (0.876) took the second place and BMI (0.761) located lastly. Conclusions:Both VAI and LAP have good prediction ability for MAFLD in the population without overweight/obesity. However, compared with VAI, LAP has higher accuracy on predicting MAFLD in the population without overweight/obesity.
3.Expression and clinical significance of autophagy-related proteins LC3, P62 and Beclin-1 in hypertrophic scar formation
Xiaoxia CHEN ; Siwei QU ; Xiao HAN ; Zizi CHEN ; Jia CHEN ; Ping LI ; Chaoqi YIN ; Sainan ZENG ; Aijun WANG ; Bin HE ; Honghui SU ; Jianda ZHOU
Journal of Chinese Physician 2019;21(4):489-494
Objective To explore the differences in autophagic expression levels between hypertrophic scar (HS) tissue and normal skin tissue,and further investigate the relationship between hypertrophic scar formation and autophagy protein expression through the rabbit ear hypertrophic scar model.Methods 30 patients with hypertrophic scar were collected.One hypertrophic scar tissue and one normal skin tissue were harvested.The relative expressions of LC3,P62 and Beclin-1 in each tissue specimen were detected by immunohistochemistry and Western blot.Western blot was used to detect the autophagic-associated protein LC3 (MAPLC3),P62 and Beclin-1 in the hypertrophic scar tissue of rabbit ear and the corresponding normal tissue of rabbit ears at 4 weeks,8 weeks,12 weeks,and 24 weeks,and further explore their clinical significance.Results In vivo,the expression of hypertrophic scar tissue protein LC3 and Beclin-1 was significantly stronger than that in normal skin tissue (P < 0.05).The expression of P62 was significantly weaker than that in normal skin tissue (P < 0.05).In animal experiments,during the process of HS formation,the protein expression of LC3 gradually increased,while the protein expression of P62 gradually decreased;the protein expression of Beclin-1 was higher than that of normal rabbit ears tissue,with statistically significant differences (P < 0.05).Conclusions The expression of LC3 and Beclin-1 in human hypertrophic scar tissues is higher than that in normal tissues.While the expression of P62 is lower than that in normal tissues.That is,the expression of autophagy in human hypertrophic scar tissue showed an upward trend in a certain period of time,and was significantly higher than that in normal tissue.
4.Anterior superior malleolus flap and second toe transplantation for thumb reconstruction with soft tissue and first web defect
Xixun WANG ; Danyu TAO ; Xuhui CHEN ; Jun LI ; Gaomeng ZHANG ; Zhenghua SHU ; Yue PAN ; Wenlang LUO ; Chaoqi DING
Chinese Journal of Microsurgery 2019;42(2):120-124
Objective To explore the clinical efficacy of anterior superior malleolus flap and second toe transplantation for thumb reconstruction and soft tissue defect repair or first web reconstruction.Methods A total of 13 cases were underwent anterior superior malleolus flap and second toe transplantation for thumb reconstruction and soft tissue defect repair from June,2008 to January,2016.There were 9 males and 4 females.The average age was 25 years old,ranging from 18 to 52.There were 3 cases of Ⅲ-degree thumb defect with soft tissue defect of hand,6 cases of Ⅳ-degree thumb defect with soft tissue defect of hand,3 cases of single Ⅴ-degree thumb defect and 1 Ⅴ-degree thumb defect with soft tissue defect.There were 5 patients accompanied with injury of other fingers of the affected hand.And the thenar muscles were also damaged to a varied degree.There were 10 patients with soft tissue defect before surgery,with the wound surface ranging from 3 cm × 5 cm-6 cm × 13 cm.And the wound of other 3 cases was closed.After the surgery,follow-up visit was conducted on the appearance,sensation,function,daily life and working condition of reconstructed thumb,the appearance and function condition of donor site.Results The reconstructed thumbs and anterior superior malleolus flaps in 13 cases survived and healed primarily.The average post-operative followed-up time was 5.5 years,ranging from 2.5 to 10.0 years.The appearance of reconstructed thumb was satisfactory,and the temperature sensation,pain sensation and touch sensation were recovered.The two-points discrimination was 8-18 mm.Moreover,the flexion and extension function,opposing function and finger-to-thumb function was recovered,which could meet the demand of daily life and work.The appearance of donor site was satisfactory,without claudication,pain or dysfunction.The survival of transplanted skin area in the calf was good,with no ulcer noticed.According to the Trial Standard for Thumb and Finger Reconstitution Functional Assessment of Hand Surgery Society of Chinese Medical Association,there were 10 cases excellent results,2 good and 1 fair.Conclusion Application of anterior superior malleolus flap and second toe transplantation for first-stage thumb reconstruction and soft tissue defect repair or thenar muscles and first web reconstruction is an effective therapeutic approach.
5.The value of urodynamic examination in treating the patient with prostatic hyperplasia accompany with dysfunction of the bladder
Huichong YU ; Dahai YU ; Chaoqi WANG
Chinese Journal of Urology 2018;39(5):382-385
Objective To investigate the value of urodynamic in treating those patients with benign prostatic hyperplasia complicated with bladder dysfunction.Methods 130 patients with benign prostatic hyperplasia were retrospectively reviewed between December 2015 and December 2016.The exclusive criteria included nervous system diseases,moderate to severe diabetes,moderate to severe hypertension,urethral stricture,bladder stones,prostate cancer,etc.The patient's age is ranged 51 to 92,mean 72 years old.The previous history of disease ranged from 3 months to 15 years,mean 4 years.All patients accepted the urodynamic examination.Different treatments were considered according to urodynamic studies.Results According to the classification of bladder outlet obstruction,130 cases were classified as bladder outlet obstruction in 109 cases,bladder suspicious obstruction in 15 cases,no bladder obstruction in 6 cases.According to the classification of bladder function,130 cases were divided into unstable bladder in 6 cases,low compliance bladder in 7 cases,bladder atony in 15 cases,bladder without tension in 3 cases,normal bladder function in 99 cases.3 cases without bladder tension,including BOO in 2 cases,suspected BOO in 1 case,were accepted the cystostomy.15 cases with less detrusor contraction,including BOO in 4 cases,suspected BOO in 9 cases,were accepted the cystostomy and TURP.Conservative treatment was performed in other 2 cases without obstruction.In 6 cases with unstable bladder,2 cases without obstruction accepted the conservative treatment,4 cases with suspected BOO accepted the TURP.In 7 patients with low compliance bladder,2 cases without obstruction accepted the conservative treatment,5 cases with suspected BOO accepted the TURP.The duration of following up was 3-6 months.The symptoms of the patients in this group improved significantly during following up.Conclusion For patients with benign prostatic hyperplasia complicated with bladder dysfunction,the results of urodynamics can help to choose the right treatment and improve effect of treatment.
6.Effects of total saponins of Panax japonicuson liver inflammation natural aging rats
Tingting XIANG ; Changcheng ZHANG ; Chaoqi LIU ; Ting WANG ; Chengfu YUAN ; Cheng XU ; Ding YUAN
Chinese Pharmacological Bulletin 2017;33(6):848-853
Aim To study the effects of total saponins of Panax japonicus(TSPJ) on liver inflammation of natural aging rats.Methods The experimental rats were allocated into seven groups (twelve rats in each group): three months group, nine months group, fifteen months group, twenty-four months group, and TSPJ low-, mid-and high-dose groups(10, 30, 60 mg·kg-1).When the rats were eighteen months old, the TSPJ low-, mid-and high-dose groups of rats were given lavage treatments with TSPJ 10,30, 60 mg·kg-1 respectively until twenty-four months.During lavage, we stopped a day every week for six consecutive months.HE staining was used to observe the pathological morphological changes.Western blot was utilized to test IL-1β and TNF-α protein expressions.RT-PCR method was adopted to test IL-1β, IL-6, IL-12, IL-17α, TNF-α, IFN-γ mRNA expressions.Results HE staining observation showed that as the rats grew older the hepatic cord and sinusoid were arranged in more severe disorder, and the fat vacuole and inflammatory cells were increased significantly.While every dose group of TSPJ could improve these pathological changes distinctly.The IL-1β, TNF-α protein and IL-1β, IL-6, IL-12, IL-17α, TNF-α, IFN-γ mRNA expressions were increased gradually as the rats grew older, and every dose group of TSPJ could reduce their expressions to some extent.Conclusion TSPJ could protect the aging rat liver to some extent by inhibiting the liver inflammation.
7.Comparative study of VAC in the preoperative and postoperative treatment of bedsore united with skin flap
Chaoqi YIN ; Zhixing KANG ; Chengqun LUO ; Ping LI ; Jia CHEN ; Shaohua WANG ; Jianda ZHOU
Journal of Chinese Physician 2017;19(5):644-646,649
Objective To investigate the clinical efficacy and mechanism of vacuum-assisted closure (VAC) in the preoperative and postoperative treatment of bedsore united with skin flap.Methods Twenty two cases with bedsore were randomly divided into experimental and control groups.In the control group,the surgery of flap was performed after the treatment of continuous negative pressure about 7-10 days and the VAC was not applied after operation.While in the experimental group,VAC was not used before operation.It was applied on flaps as soon as sutured the border of flap and decubitus ulcers and removed after 7-10 days.By comparing the general appearance of two groups,microvessel count and the detection rate of bacterial culture and other indicators,the clinical effects of two treatments were investigated and the preliminary mechanism was analyzed.Results After preoperative VAC treatment,11 cases of control group showed a little granulation tissue growth,less subcutaneous hematoma and wound effusion,increased microvessel count and negative bacterial culture.However,there were 4 cases of death cavity residual,subcutaneous hematoma and wound effusion,positive bacterial culture and another 4 cases of delayed healing with skin flap repairing bedsore.The application of VAC in experimental group showed close contact of flap with the basement,less effusion,increased microvessel count and negative bacterial culture.One case of skin flap had a small area of separation,after the dressing of skin and the flap survived.The other wounds healed by first intention.Conclusions The use of VAC to repair bedsore can reduce the number of operation,and it is beneficial to the flap survival.
8.Retrospective analysis of 185 cases of skin ulcers
Xiangqin HUANG ; Jia CHEN ; Shaohua WANG ; Cheng PENG ; Ping LI ; Chaofei HAN ; Jianfei ZHANG ; Chaoqi YIN ; Jianda ZHOU
Journal of Chinese Physician 2017;19(5):659-663
Objective To explore the clinical features and treatment of skin ulcers.Methods A total of 185 skin ulcer patients with clinical data was analyzed retrospectively in our hospital from October 2014-October 2016.Results Of the 185 patients with skin ulcers,84(45.4%) were 51-70 years old,diabetic ulcers were most common,and most occurred in ankle (44.3%) and limbs (35.7%).After further statistical analysis,the healing rate of patients treated by surgery was significantly higher than non-surgical patients.Surgical procedures of local flap transfer after ulcer resection were most commonly used and of the highest healing rate.The number of ulcers,the largest diameter of ulcers,scar ulcers and blood barrier ulcers were four factors that were independent of the outcome of skin ulcers.Conclusions Skin ulcers hospitalized patients are more common in patients over 50 years of age,occur most in the lower limbs.Surgical therapy is the preferred treatment for most ulcers,of which local flap transfer after ulcer resection has the best effect.Ulcer patients should actively treat the primary disease,strengthen the care,early diagnosis,and early treatment.
9.Anti-inflammatory effect of Chikusetsu oleanane saponin on RAW264. 7 cell through regulating SIRT1 activity
Qin YUAN ; Ding YUAN ; Zhiyong ZHOU ; Chaoqi LIU ; Ting WANG ; Tingting XIANG ; Changcheng ZHANG
Chinese Pharmacological Bulletin 2016;32(3):349-354
Aim To investigate SIRT1 activity and an-ti-inflammatory effects of Chikusetsu oleanane saponin ( COS) on the RAW264. 7 macrophage cells induced by lipopolysaccharide ( LPS ) . Methods The nitric oxide ( NO) release was detected with Griess method. Western blot was used to detect the expression of tumor necrosis factor-α( TNF-α) , interleukin 1β( IL-1β) . Nuclear factor-κB ( NF-κB) and silent information ad-justment factor 1 ( SIRT1 ) were tested by immunofluo-rescence. Results 1 mg · L-1 LPS co-cultured with COS at 25~300 mg·L-1 had no significant effect on the growth of RAW264. 7 cells. Compared with the LPS group, COS effectively inhibited the NO release and suppressed the expression of TNF-α and IL-1β, and also inhibited the translocation of NF-κB and up-regulation of SIRT1 . Conclusion COS has protective effects on RAW264. 7 cells stimulated by LPS, which may be related to up-regulating the expression of SIRT1 and promoting the deacetylation of NF-κB, thereby in-hibiting the translocation of NF-κB and reducing the expression of TNF-α and IL-1β.
10.The correlation between hepatitis B virus DNA replication and large envelope protein in serum
Chaoqi HE ; Jiangfeng QIN ; Youfang LIANG ; Tianxiang WANG ; Aihua SUN
Chinese Journal of Experimental and Clinical Virology 2015;29(1):23-25
Objective To study the significance of serum hepatitis B virus large envelope protein (HBV-LP),HBV DNA and HBeAg in the diagnosis of viral replication.Methods In 623 chronic HBV patients,HBV-LP and HBeAg were detected by ELISA and HBV DNA was quantitaively detected by realtime polymerase chain reaction(PCR).Results The positive rate of HBV-LP and HBV DNA were higher than that of HBeAg in all chronic HBV patients(x2 =22.3 and 9.4,P <0.01),and the positive rate of HBVLP and HBV DNA was no significant difference (x2 =2.8,P > 0.05),The positive rate of HBV-LP and HBV DNA detected in HBeAg(+) or HBeAg(-) were not significantly different(x2 =3.2 and 0.6,P > 0.05).HBV DNA copies and A vaule of HBV-LP was a positive correlation(r =0.874,P <0.01),HBV DNA copies of different groups was significantly different from HBV-LP A values(F =6.987,P <0.01).In 100 healthy people,the detecting results of HBV DNA and HBV-LP are negetive.Conclusion HBV-LP was better than HBeAg for detection of HBV replication.There is a good correlation between the copies of HBV DNA and the levels of HBV-LP,HBV-LP expression can reflect the replication of HBV.


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