1.IsoVISoR: Towards 3D Mesoscale Brain Mapping of Large Mammals at Isotropic Sub-micron Resolution.
Chao-Yu YANG ; Yan SHEN ; Xiaoyang QI ; Lufeng DING ; Yanyang XIAO ; Qingyuan ZHU ; Hao WANG ; Cheng XU ; Pak-Ming LAU ; Pengcheng ZHOU ; Fang XU ; Guo-Qiang BI
Neuroscience Bulletin 2025;41(2):344-348
2.The neuroprotective effect of Wenfei Jiangzhuo formula on vascular dementia model rats based on regulation of mitochondrial homeostasis by PGAM5-Drp1 axis
Ding ZHANG ; Zhi-Han HU ; Chun-Ying SUN ; Xiao-Dong ZHU ; Fang-Cun LI ; Ming-He JIANG ; Hong-Ling QIN ; Wei CHEN ; Yue-Qiang HU
Chinese Pharmacological Bulletin 2024;40(11):2158-2164
Aim To observe the effects of Wenfei Jiangzhuo formula(WFJZF)on rats with vascular de-mentia and investigate its possible mechanism of ac-tion.Methods Thirty-six healthy male SD rats were randomly divided into the sham group,model group,donepezil group,and low-dose,medium-dose and high-dose groups of Wenfei Jiangzhuo formula,with six rats per group.Except for the sham group,the other groups were prepared as VaD models,and each group was gavaged with the corresponding drugs after suc-cessful modeling,and tests were performed after three weeks of treatment.Behavioral,hippocampal CA1 area morphology,neural dendrites and mitochondrial chan-ges were observed in all groups of rats,and phospho-glycerate mutase 5(PGAM5),dynamics-related pro-teins1(Drp1),opticatrophyprotein-1(OPA1),and other proteins were detected in each group.Results Compared with the sham group,rats in the model group and each intervention group had prolonged es-cape latency(P<0.05),a shorter number of travers-als across the platforms(P<0.05),a sparse morphol-ogy of hippocampal neurons,a reduction in the number of secondary dendritic spines,and a rupture of the out-er membrane of the mitochondria;the expression of the PGAM5 and Drp1 proteins in hippocampal tissues was elevated(P<0.05),and the expression of the OPA1 and Mfn1/2 protein expression decreased(P<0.05);compared with the model group,donepezil group and Wenfei Jiangzhuo formula high-dose group of rats had shorter evasion latency(P<0.05),increased number of times to traverse the platform(P<0.05),increased number of hippocampal neurons,tightly packed,more secondary dendritic structures,and reduced mitochon-drial damage;the expression of PGAM5 and Drp1 pro-teins was reduced(P<0.05),and the expression of OPA1 and Mfn1/2 proteins was elevated(P<0.05).Conclusions Wenfei Jiangzhuo formula can regulate the PGAM5-Drp1 signaling axis to improve the balance of mitochondrial homeostasis,thus improving the cog-nitive condition of the brain and exerting cerebroprotec-tive effects.
3.Predictive value of 18F-FDG PET/CT in molecular subtyping for triple-negative breast cancer
Jianjing LIU ; Haiman BIAN ; Qiang FU ; Ziyang WANG ; Fang YANG ; Dong DAI ; Wei CHEN ; Lei ZHU ; Wengui XU
Chinese Journal of Radiological Medicine and Protection 2024;44(5):421-427
Objective:To explore the predictive value of 18F-FDG PET/CT in molecular subtyping of triple-negative breast cancer. Methods:A retrospective analysis was performed on the clinical and imaging data of 227 breast cancer patients who underwent 18F-FDG PET/CT examination in the Tianjin Medical University Cancer Institute & Hospital from January 1, 2010 to December 31, 2022. Based on the expression levels of estrogen receptor (ER), progesterone receptor (PR), and human epidermal growth factor receptor 2 (HER-2) in the primary breast cancer, the patients were categorized into two groups: triple-negative breast cancer (TNBC) and non-TNBC. Radiomic features were extracted from images of both groups, and a radiomic model was constructed to predict the molecular subtype of the TNBC groups. In addition, the clinical data, CT morphological features, and PET metabolic parameters of both groups were compared to determine the indicators with statistically significant differences and develop a comprehensive radiomic model combined with clinical characteristics. Results:Compared to the non-TNBC group, the TNBC groups exhibited more significant invasiveness in terms of tumor diameter, margins, ipsilateral axillary lymph node metastasis, invasion of neighboring skin or papillae, and PET metabolic parameters ( t = -3.19; χ2 = 7.30, 8.10, 5.34; t = 3.80, 3.30, 3.42, P < 0.05). The constructed 18F-FDG PET/CT radiomic model proved effective in predicting the molecular subtype of the TNBC group, and the receiver operating characteristic (ROC) curve showed an area under the curve (AUC) of 0.83 (95% CI 0.78-0.88), an accuracy of 75.9%, a sensitivity of 74.5%, and a specificity of 77.2%. In contrast, the constructed comprehensive radiomic model displayed an AUC of 0.86 (95% CI 0.81-0.90), an accuracy of 77.2%, a sensitivity of 78.6%, and a specificity of 75.9%. Conclusions:18F-FDG PET/CT plays an important role in predicting molecular subtypes of TNBC. The constructed radiomic model and comprehensive radiomic model can further enhance the prediction efficacy of PET metabolic parameters and accelerate the development of accurate treatment protocols in clinical practice, thus improving the prognosis of breast cancer.
4.Based on the concept and technique of full reconstruction of the hallux tissue flap treatment for thumb and fingertip defect reconstruction
Jie FANG ; Hui ZHU ; Guiqian LIU ; Shuo XU ; Qiang QI ; Wei ZHANG ; Weiya QI ; Dawei ZHENG ; Chao CHEN
Chinese Journal of Plastic Surgery 2024;40(1):69-75
Objective:To investigate the clinical efficacy of reconstruction with the hallux tissue flap for the fingertip defects of thumb and fingers based on the concept and technique of full reconstruction.Methods:From September 2022 to February 2023, the patients with thumb and fingertip defects who were reconstructed using the concept and technology of full reconstruction in Xuzhou Renci Hospital were respective analysised. Based on the degree of defect in the nail bed, bone, and soft tissue of the hand, a hallux osteo-onychocutaneous flap or hallux nail flap was designed and harvested from the same side of the toe, and free transplantation was performed to reconstruct the damaged fingertip. The wounds in the donor site that could not be sutured primarily were treated with dressing change. The surgical complications, the appearance of the reconstructed finger and donor site, Semmes-Weinstein monofilament (SWM) examination, 2-point discrimination (2PD) measurement, and patient satisfaction were regularly recorded. According to the functional evaluation criteria for thumb and finger reconstruction issued by the Hand Surgery of the Chinese Medical Association, the function of the reconstructed fingers was evaluated as excellent (13-15 points), good (9-12 points), fair (5-8 points), and poor (≤4 points). The patients were investigated whether they were satisfied with the outcomes.Results:A total of 8 patients (9 fingers) including 5 males and 3 females were enrolled. The average age was 32.4 years (range, 22-46 years). There were 1 thumb, 2 index fingers, 5 middle fingers, and 1 ring finger. 7 cases (8 fingers) were repaired with the hallux osteo-onychocutaneous flap, and 1 case (1 finger) was repaired with the hallux nail flap. All the reconstructed fingers survived uneventfully, and the recipient site healed primarily, and the donor site healed after dressing change, without infection and other complications. All the 8 patients (9 fingers) were followed up for 3-7 months. The nail plate of the donor site naturally grew and recovered nearly normal. Depression formation was observed at part of the deck junction of the recipient site, and scars were left in the pulp and fibular side of the hallux. At the final follow-up, the reconstructed fingers recovered protective sensation and tactile sense, and the 2PD of the reconstructed fingers was 9 mm and 10 mm in two cases. The SWM score was 3.60-4.31, mean 3.96. The results were the monofilament specifications and indicating the protective sensation was diminished. Functional evaluation of reconstructed fingers: 9 fingers in 8 cases scored 13-14 points, all reaching an excellent level. All patients were satisfied with the surgical results.Conclusion:Based on the concept and technology of full reconstruction, the fingertip defects of the thumb and finger can recover close to normal with the hallux tissue flap in the short-term, while ensuring the overall shape and function of the hallux donor area, achieving a balance between the donor and recipient areas, and achieving satisfactory clinical results.
5.Amplitude-integrated electroencephalography monitoring results of hospitalized neonates in plateau areas
Xiao-Fen ZHAO ; Guo-Qiang CHENG ; Peng-Na ZHAO ; Mei ZHAO ; Shuang-Yan ZHU ; Yang-Fang LI ; Wen-Hao ZHOU
Chinese Journal of Contemporary Pediatrics 2024;26(8):817-822
Objective To investigate the amplitude-integrated electroencephalography(aEEG)monitoring results of hospitalized neonates in plateau areas.Methods A retrospective analysis was conducted on 5 945 neonates who were admitted to the Department of Neonatology,Kunming Children's Hospital,and received aEEG monitoring from January 2020 to December 2022.According to the aEEG monitoring results,they were divided into a normal aEEG group and an abnormal aEEG group.The incidence rate of aEEG abnormalities was analyzed in neonates with various systemic diseases,as well as the manifestations of aEEG abnormalities and the consistency between aEEG abnormalities and clinical abnormalities.Results Among the 5 945 neonates,the aEEG abnormality rate was 19.28%(1 146/5 945),with an abnormality rate of 29.58%(906/3 063)in critically ill neonates and 8.33%(240/2 882)in non-critically ill neonates(P<0.05).The children with inherited metabolic diseases showed the highest aEEG abnormality rate of 60.77%(79/130),followed by those with central nervous system disorders[42.22%(76/180)]and preterm infants[35.53%(108/304)].Compared with the normal aEEG group,the abnormal aEEG group had significantly lower age and gestational age,as well as a significantly lower birth weight of preterm infants(P<0.05).Among the 1 146 neonates with aEEG abnormalities,the main types of aEEG abnormalities were sleep cycle disorders in 597 neonates(52.09%),background activity abnormalities in 294 neonates(25.65%),and epileptiform activity in 255 neonates(22.25%),and there were 902 neonates(78.71%)with abnormal clinical manifestations.The sensitivity and specificity of aEEG monitoring for brain function abnormalities were 33.51%and 92.50%,respectively.Conclusions In plateau areas,there is a relatively high rate of aEEG abnormalities among hospitalized neonates,particularly in critically ill neonates and those with smaller gestational ages and younger ages,suggesting a high risk of brain injury.Therefore,routine aEEG monitoring for the hospitalized neonates can help with the early detection of brain function abnormalities,the decision-making in treatment,and the formulation of brain protection strategies.
6.Based on the concept and technique of full reconstruction of the hallux tissue flap treatment for thumb and fingertip defect reconstruction
Jie FANG ; Hui ZHU ; Guiqian LIU ; Shuo XU ; Qiang QI ; Wei ZHANG ; Weiya QI ; Dawei ZHENG ; Chao CHEN
Chinese Journal of Plastic Surgery 2024;40(1):69-75
Objective:To investigate the clinical efficacy of reconstruction with the hallux tissue flap for the fingertip defects of thumb and fingers based on the concept and technique of full reconstruction.Methods:From September 2022 to February 2023, the patients with thumb and fingertip defects who were reconstructed using the concept and technology of full reconstruction in Xuzhou Renci Hospital were respective analysised. Based on the degree of defect in the nail bed, bone, and soft tissue of the hand, a hallux osteo-onychocutaneous flap or hallux nail flap was designed and harvested from the same side of the toe, and free transplantation was performed to reconstruct the damaged fingertip. The wounds in the donor site that could not be sutured primarily were treated with dressing change. The surgical complications, the appearance of the reconstructed finger and donor site, Semmes-Weinstein monofilament (SWM) examination, 2-point discrimination (2PD) measurement, and patient satisfaction were regularly recorded. According to the functional evaluation criteria for thumb and finger reconstruction issued by the Hand Surgery of the Chinese Medical Association, the function of the reconstructed fingers was evaluated as excellent (13-15 points), good (9-12 points), fair (5-8 points), and poor (≤4 points). The patients were investigated whether they were satisfied with the outcomes.Results:A total of 8 patients (9 fingers) including 5 males and 3 females were enrolled. The average age was 32.4 years (range, 22-46 years). There were 1 thumb, 2 index fingers, 5 middle fingers, and 1 ring finger. 7 cases (8 fingers) were repaired with the hallux osteo-onychocutaneous flap, and 1 case (1 finger) was repaired with the hallux nail flap. All the reconstructed fingers survived uneventfully, and the recipient site healed primarily, and the donor site healed after dressing change, without infection and other complications. All the 8 patients (9 fingers) were followed up for 3-7 months. The nail plate of the donor site naturally grew and recovered nearly normal. Depression formation was observed at part of the deck junction of the recipient site, and scars were left in the pulp and fibular side of the hallux. At the final follow-up, the reconstructed fingers recovered protective sensation and tactile sense, and the 2PD of the reconstructed fingers was 9 mm and 10 mm in two cases. The SWM score was 3.60-4.31, mean 3.96. The results were the monofilament specifications and indicating the protective sensation was diminished. Functional evaluation of reconstructed fingers: 9 fingers in 8 cases scored 13-14 points, all reaching an excellent level. All patients were satisfied with the surgical results.Conclusion:Based on the concept and technology of full reconstruction, the fingertip defects of the thumb and finger can recover close to normal with the hallux tissue flap in the short-term, while ensuring the overall shape and function of the hallux donor area, achieving a balance between the donor and recipient areas, and achieving satisfactory clinical results.
7.Investigation of curative effect of various types of Flow-through flaps in reconstruction of wounds with defect of palmar proper digital artery
Jie FANG ; Hui ZHU ; Weiya QI ; Song ZHANG ; Wei ZHANG ; Qiang QI ; Dawei ZHENG
Chinese Journal of Microsurgery 2024;47(3):300-306
Objective:To investigate the therapeutic efficiency of various types of Flow-through flap in the reconstruction of soft tissue defect with defect of proper palmar digital artery.Methods:Retrospective analysis was carried out on 22 patients who were treated with perforator flap of posterior interosseous artery, venous flap, the superficial palmar branch of radial artery flap, and "C" shaped flap of the great toe for soft tissue defects of hands with defects of palmar proper digital artery in the Department of Hand Surgery of Xuzhou Renci Hospital from January 2019 to January 2023. The patients were 15 males and 7 females with an average age of 39.5 (21-61) years old. The injured digits were 1 thumb, 6 index fingers, 8 middle fingers, 5 ring fingers and 2 little fingers. Ten injuries were of left hand and 12 of right hand. The wound size was 1.5 cm × 1.5 cm-4.5 cm × 3.0 cm, and the flap size was 2.0 cm × 1.5 cm-5.0 cm × 4.0 cm. The donor site wound were directly sutured and closed or treated by dressing changes to heal. Postoperative follow-ups were arranged at 1, 2, 3, 6 and 12 months after surgery at outpatient clinic, through telephone or WeChat interviews. The flap survival, postoperative complications at donor and recipient sites, patient satisfaction and range of motion of interphalangeal joint were observed and recorded.Results:Twenty flaps survived uneventfully. Tension vesicles appeared on the 2 arterialised venous flaps and healed after dressing changes. All donor sites healed uneventfully. All patients were included in the postoperative follow-up for an average of 10.3 (6-24) months. Appearance of the perforator flaps of posterior interosseous artery was bloated and the movement of interphalangeal joint was limited in 1 digit, however the patient refused a further flap thinning surgery. Scar dissection was performed on 1 of the venous flaps in stage II surgery. The rest of 20 patients were satisfied with the appearance of flaps hence no further thinning surgery was performed. According to Evaluation Trial Standards of Upper Limb Partial Functional of Hand Surgery of Chinese Medical Association, 12 cases were evaluated as excellent, 9 cases as good, and 1 case as poor at the last follow-up. The mean static TPD was found at 6.9 (5-11) mm, and the average Semmes-Weinstein monofilament (SWM) test was found at 4.47 (3.61-6.65).Conclusion:The 4 types of perforator flap can be employed in the reconstruction of digital defect with defect of palmar proper digital artery in a one-stage surgery with satisfactory outcomes. Surgeons can accordingly make individualised surgical plans as required for the patients.
8.Manipulation treatment of lumbar disc herniation based on the model of muscles and bones assessment.
Bin XUE ; Tao LIU ; Hong ZHU ; Fei-Yue LI ; Jia-Yu ZHANG ; Qiang WANG ; Xiao-Bing XI ; Ling-Jun KONG ; Xin ZHOU ; Qing-Guang ZHU ; Min FANG
China Journal of Orthopaedics and Traumatology 2024;37(11):1080-1086
OBJECTIVE:
To explore the effectiveness of manipulation treatment for lumbar disc herniation (LDH) based on the model of muscle and bone assessment.
METHODS:
From May 2022 to August 2023, using the methods single-center randomized controlled in Ruijin Hospital Affiliated to Shanghai Jiaotong University School of Medicine, 72 patients were treated with LDH and divided into muscle and bone assessment model manipulation group and the two step seven gimmick group according to the random number table method, the muscle and bone assessment model manipulation group fall off in 1 case, the two step seven gimmick group falls off in 2 cases. There were 35 cases in the muscle and bone assessment model manipulation group, including 12 males and 23 females;The age was 27 to 48 years old with an average of (37.77±7.63) years old. The course of disease was 35 to 180 days with an average of (83.68±69.01) days. The patients were treated with manual therapy under the guidance of muscle and bone assessment model, twice a week for 4 weeks. There were 34 cases in the two step seven gimmick group including 12 males and 22 females;The age was 26 to 49 years old with an average of (37.59±7.43) years old;The course of disease was 40 to 175 days with an average of (82.15±68.87) days. The patients were treated with two step seven gimmick method, 2 times a week, for 4 weeks. The visual analogue scale (VAS) and Oswestry disability index (Oswestry disability index, ODI) questionnaire, muscle tension and lumbar spine angle and the straight leg-raising activities were compared between two groups before and 4 weeks after treatment.
RESULTS:
The VAS of the muscle and bone assessment model manipulation group and the two step seven gimmick group(6.51±0.61) and (6.62±0.56) before treatment decreased to 2.40±0.81 and 3.18±0.78 after 4 weeks of treatment, respectively, and the muscle and bone assessment model manipulation group was significantly lower than the two step seven gimmick group (P<0.01). The ODI of the muscle and bone assessment model manipulation group and the two step seven gimmick group were (64.57±5.11) and (65.02±5.18) before treatment, decreased to (18.60±2.27) and (24.70±2.14) after 4 weeks of treatment, and the ODI of the muscle and bone assessment model manipulation group was significantly lower than that of the two step seven gimmick group (P<0.01). Before the treatment, side erector spinae, gluteus medius, and gastrocnemius muscle tension were (59.95±2.60), (62.59±2.51), (49.97±2.01) in the muscle and bone assessment model manipulation group and (60.39±3.84), (62.47±3.27), (49.55±1.27) in the two step seven gimmick group;After 4 weeks of treatment, the muscle tension of erector spinae, gluteus medius and gastrocnemius on the affected side were (56.58±2.71), (60.44±2.31) and (49.19±1.57) in the muscle and bone assessment model manipulation group, (58.28±3.79), (60.11±2.87), (48.55±0.90) in the two step seven gimmick group, the differences had statistical significance before and after treatment of two groups(P<0.01). The muscle and bone assessment model manipulation group was better than the two step seven gimmick group in improving the erector spinae muscle tension on the affected side (P<0.05), and there was no significant difference in the rest (P>0.05). Before the treatment, lumbar proneness, stretch, subject to lateral flexion and lateral angle of the straight leg-raising on the affected side were (46.00±8.89)°, (13.57±3.75)°, (12.29±3.50) °, (43.71±7.98) ° in the muscle and bone assessment model manipulation group, (45.14±6.24) °, (12.23±3.75) °, (12.66±2.98) ° and (44.18±3.50) ° in the two step seven gimmick group. After 4 weeks of treatment, the angles of lumbar flexion, extension, flexion on the affected side and straight leg raising on the affected side were (76.29±4.43) °, (20.00±1.71) °, (22.43±2.81) °, (70.41±7.59) ° in the muscle and bone assessment model manipulation group, and (75.75±6.38) °, (16.43±3.36) °, (20.19±3.52) °, (65.42±6.15) ° in the two step seven gimmick group. The difference had statistical significance before and after treatment in two groups(P<0.01), a comparison between groups, after 4 weeks of treatment, the angles of lumbar flexion and extension, affected side flexion, and lower limb straight leg elevation in the muscle and bone assessment model manipulation group were better than those in the two step seven gimmick group (P<0.05). Before the treatment, pelvic tilt, lumbar lordosis angle were (2.71±1.01) mm, (37.63±3.35) ° in the muscle and bone assessment model manipulation group, and (2.69±0.97) mm, (36.98±3.73) ° in the two step seven gimmick group;After 4 weeks of treatment, the pelvic tilt and lumbar lordosis angle were (0.84±0.36) mm and (41.64±2.96) ° in the muscle and bone assessment model manipulation group, and those in the method of two step seven gimmick group were (1.18±0.75) mm and (41.70±3.14) °. There were significant differences before and after treatment in both groups (P<0.01), and the improvement of pelvic tilt in the muscle and bone assessment model manipulation group was better than that in the method of two step seven gimmick group after 4 weeks of treatment (P<0.05).
CONCLUSION
The manipulation under the guidance of the muscle and bone assessment model can effectively improve the pain and dysfunction of LDH patients, and has a better effect than the two-step seven-method manipulation group in improving the muscle tension, lumbar motion function and posture.
Humans
;
Male
;
Female
;
Intervertebral Disc Displacement/physiopathology*
;
Middle Aged
;
Adult
;
Lumbar Vertebrae
9.Clinical observation of acupuncture at Huiyin (CV 1) for chronic severe functional constipation.
Li-Fang JIANG ; Jie WU ; Qiang FU ; Li-Hong JIANG ; Chen CHEN ; Dan ZHU ; Yuan-Ming ZHONG
Chinese Acupuncture & Moxibustion 2023;43(2):128-132
OBJECTIVE:
To compare the effect of acupuncture at Huiyin (CV 1) and oral administration of western medication in treatment of chronic severe functional constipation (CSFC).
METHODS:
A total of 64 patients with CSFC were randomly divided into an acupuncture group (32 cases, 5 cases dropped off) and a western medication group (32 cases, 4 cases dropped off). Both groups were given routine basic treatment. The acupuncture group was treated by directly puncture of 20-30 mm at Huiyin (CV 1), once a day for the first 4 weeks, 5 times a week, once every other day for the next 4 weeks, 3 times a week, totally for 8 weeks. The western medication group was treated with 2 mg prucalopride succinate tablets orally before breakfast every day for 8 weeks. The average number of weekly spontaneous bowel movement (SBM) of the two groups were observed before treatment and 1-8 weeks into treatment. The constipation symptom score before and after treatment, and in follow-up of 1 month after treatment, as well as quality of life [patient assessment of constipation quality of life questionnaire (PAC-QOL) score and the proportion of patients of PAC-QOL score difference before and after treatment≥1] before and after treatment were compared in the two groups. The clinical effects of the two groups were evaluated after treatment and in follow-up.
RESULTS:
Compared before treatment, the average number of weekly SBM in the two groups was increased 1-8 weeks into treatment (P<0.05). The average number of weekly SBM in the acupuncture group was less than that in the western medication group 1 week into treatment (P<0.05), and the average number of weekly SBM in the observation group was more than that in the western medication group 4-8 weeks into treatment (P<0.05). The scores of constipation symptom after treatment and in follow-up and scores of PAC-QOL after-treatment in the two groups were lower than those before treatment (P<0.05), and those in the acupuncture group were lower than the western medication group (P<0.05). The proportion of patients of PAC-QOL score difference before and after treatment≥1 in the acupuncture group was higher than that in the west medication group (P<0.05). The total effective rates after treatment and in follow-up in the acupuncture group were 81.5% (22/27) and 78.3% (18/23), respectively, which were better than 42.9% (12/28) and 43.5% (10/23) in the western medication group (P<0.05).
CONCLUSION
Acupuncture at Huiyin (CV 1) can effectively increase the number of spontaneous defecation in patients with CSFC, reduce constipation symptoms, improve the quality of life, and the effect after treatment and in follow-up is better than oral western medication.
Humans
;
Quality of Life
;
Treatment Outcome
;
Acupuncture Points
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Constipation/therapy*
;
Acupuncture Therapy
10.scRNA-seq reveals that origin recognition complex subunit 6 regulates mouse spermatogonial cell proliferation and apoptosis via activation of Wnt/β-catenin signaling.
Shi-Wei LIU ; Jia-Qiang LUO ; Liang-Yu ZHAO ; Ning-Jing OU ; CHAO-YANG ; Yu-Xiang ZHANG ; Hao-Wei BAI ; Hong-Fang SUN ; Jian-Xiong ZHANG ; Chen-Cheng YAO ; Peng LI ; Ru-Hui TIAN ; Zheng LI ; Zi-Jue ZHU
Asian Journal of Andrology 2023;26(1):46-56
The regulation of spermatogonial proliferation and apoptosis is of great significance for maintaining spermatogenesis. The single-cell RNA sequencing (scRNA-seq) analysis of the testis was performed to identify genes upregulated in spermatogonia. Using scRNA-seq analysis, we identified the spermatogonia upregulated gene origin recognition complex subunit 6 (Orc6), which is involved in DNA replication and cell cycle regulation; its protein expression in the human and mouse testis was detected by western blot and immunofluorescence. To explore the potential function of Orc6 in spermatogonia, the C18-4 cell line was transfected with control or Orc6 siRNA. Subsequently, 5-ethynyl-2-deoxyuridine (EdU) and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assays, flow cytometry, and western blot were used to evaluate its effects on proliferation and apoptosis. It was revealed that ORC6 could promote proliferation and inhibit apoptosis of C18-4 cells. Bulk RNA sequencing and bioinformatics analysis indicated that Orc6 was involved in the activation of wingless/integrated (Wnt)/ β-catenin signaling. Western blot revealed that the expression of β-catenin protein and its phosphorylation (Ser675) were significantly decreased when silencing the expression of ORC6. Our findings indicated that Orc6 was upregulated in spermatogonia, whereby it regulated proliferation and apoptosis by activating Wnt/β-catenin signaling.

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