1.Effect and Mechanism of Nongsuo Dangguiwan in Relieving Oxidative Stress in Premature Ovarian Insufficiency Rats Based on Nrf2/HO-1 Signaling Pathway
Jiaqi LUO ; Xiaoli DU ; Dongmei CHEN ; Miao CHEN ; Na HU ; Shasha XING ; Wenzhao LIU ; Ruiying TIAN ; Li YANG ; Jing WANG ; Rui HE ; Huiming MA
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(13):1-9
ObjectiveTo investigate the effect and mechanism of Nongsuo Dangguiwan in improving ovarian oxidative stress in rats with ovarian dysfunction. MethodThirty-six adult female SD rats were randomly divided into normal group, model group, positive drug group (Femoston, 0.3 mg·kg-1), and high-, medium-, and low-dose groups of concentrated Nongsuo Dangguiwan (2.08, 4.16, 8.32 g·kg-1), with six rats in each group. Rats, except for those in the normal group, were injected with 80 mg·kg-1 vinyl cyclohexene dioxide (VCD) per day for 14 consecutive days to induce ovarian dysfunction. From the 15th day, rats were treated with corresponding drugs by gavage, while those in the model group received 2 mL·kg-1 saline, once daily for 28 consecutive days. The ovarian index, levels of related hormones including estradiol (E2), follicle-stimulating hormone (FSH), luteinizing hormone (LH), and anti-mullerian hormone (AMH) in serum, as well as superoxide dismutase (SOD) activity and glutathione (GSH) content in serum were measured by enzyme-linked immunosorbent assay (ELISA). The malondialdehyde (MDA) content in serum was detected by the thiobarbituric acid (TAB) method. Ovarian morphology was observed by hematoxylin-eosin (HE) staining. The expression of nuclear factor E2-related factor 2 (Nrf2), heme oxygenase-1 (HO-1), SOD2, and SOD1 in ovarian tissues was detected by immunohistochemistry (IHC) and Western blot. ResultCompared with the normal group, the model group showed a significant reduction in growing follicles in the ovary, loose arrangement of granulosa cells in the follicle, decreased body weight, ovarian index, and serum AMH and E2 levels, increased LH and FSH levels (P<0.01), reduced levels of SOD and GSH in serum (P<0.01), and increased MDA level (P<0.01). Compared with the model group, the groups with drug intervention showed increased ovarian index (P<0.05, P<0.01), increased serum E2 level (P<0.05, P<0.01), decreased FSH, AMH, and LH levels (P<0.05, P<0.01), increased number of growing follicles in the ovary, potentiated SOD activity in serum, increased GSH content, decreased MDA content (P<0.05, P<0.01), and up-regulated expression levels of Nrf2, HO-1, SOD2, and SOD1 proteins in ovarian tissues (P<0.05, P<0.01). ConclusionNongsuo Dangguiwan can regulate serum hormone levels, increase the expression of Nrf2, HO-1, SOD2, and SOD1 in ovarian tissues, and improve ovarian antioxidant capacity to resist oxidative stress injury, thereby improving ovarian reserve function.
2.Improving Effect of Jianpi Yishen Huazhuo Recipe on Ovarian Function in Rats with Polycystic Ovary Syndrome Based on Bax/Bcl-2/Caspase-3 Pathway
ZHANG Xiaojing ; CHEN Miao ; FENG Yahong ; DU Xiaoli ; JI Dejiang ; MA Huiming
Chinese Journal of Modern Applied Pharmacy 2023;40(20):2891-2896
OBJECTIVE To investigate the mechanism of Jianpi Yishen Huazhuo Recipe in regulating ovarian function in rats with polycystic ovary syndrome(PCOS) through the Bax/Bcl-2/Caspase-3 pathway. METHODS SD rats were fed with a high-fat diet combined with letrozole to establish a model of PCOS. The successfully modeled rats were randomly divided into blank control group, model group, high-, medium-, and low-dose Jianpi Yishen Huazhuo Recipe groups, as well as positive drug metformin group. ELISA was used to detect the serum levels of LH, T, and LH/FSH in each group of rats. HE staining was used to observe the morphology of ovarian tissue. Immunohistochemistry and Western blotting was used to detect the protein expression of Bax, Bcl-2, and Caspase-3 in ovarian tissues of rats. RESULTS ELISA results showed that the levels of LH, T, and LH/FSH in each dose group of Jianpi Yishen Huazhuo Recipe significantly decreased, while the level of FSH significantly increased. HE staining results showed that ovarian tissue in each dose group of Jianpi Yishen Huazhuo Recipe was significantly improved. Immunohistochemistry results showed that Bax expression increased in granulosa cells, interstitial cells, and luteal cells, while Bcl-2 expression increased in oocytes, luteal cells, and follicular fluid. Caspase-3 expression increased in granulosa cells, oocyte membranes, and interstitial cells. Western blotting results showed that the relative expression of Bax and Caspase-3 proteins decreased in each dose group of Jianpi Yishen Huazhuo Recipe, while the relative expression of Bcl-2 protein increased, and the ratio of Bax/Bcl-2 decreased. CONCLUSION Jianpi Yishen Huazhuo Recipe can improve ovarian ovulation function by downregulating the expression of Bax and Caspase-3 proteins in ovarian tissue of PCOS rats, reducing the ratio of Bax/Bcl-2, upregulating the expression of Bcl-2 protein, adjusting the process of ovarian cell proliferation and apoptosis, improving the ovarian microenvironment, regulating endocrine levels, and promoting follicular development and maturation.
3.Effect of Jianpi Yishen Huazhuo Prescription on Ovarian Function in Rats with Polycystic Ovary Syndrome Based on NLRP3 Inflammatory Pathway
Miao CHEN ; Xiaoli DU ; Yahong FENG ; Xiaojing ZHANG ; Jiaqi LUO ; Jing WANG ; Nan NAN ; Wenzhao LIU ; Shasha XING ; Ruiying TIAN ; Li YANG ; Miao SUN ; Rui HE ; Dongmei CHEN ; Huiming MA
Chinese Journal of Experimental Traditional Medical Formulae 2022;28(20):61-70
ObjectiveTo explore the mechanism of Jianpi Yishen Huazhuo prescription in the improvement of ovarian function in polycystic ovary syndrome (PCOS). MethodSeventy female SD rats in SPF grade were randomly divided into 6 groups, 15 in the blank group and 15 in the model group, 10 in the metformin group (0.1 g·kg-1·d-1), and 10 in the low (1.275 g·kg-1·d-1), medium (2.55 g·kg-1·d-1), and high-dose (5.10 g·kg-1·d-1) Jianpi Yishen Huazhuo prescription groups. The blank group was given normal saline (10 mL·kg-1·d-1) by gavage and ordinary feed, and the other groups were given letrozole (1 mg·kg-1·d-1) by gavage combined with high-fat feed for 21 days to induce the model of PCOS. After modeling, the blank group and model group were given equal volume normal saline by gavage, and each drug group was given the corresponding dose of the drug by gavage for 30 days. The changes in body mass and fasting blood glucose (FPG) of rats before and after modeling were compared. Hematoxylin eosin (HE) staining was used to observe the morphological change in the ovaries of rats in each group. The serum levels of follicle stimulating hormone (FSH), luteinizing hormone (LH), testosterone (T), anti-Mullerian hormone (AMH), and estradiol (E2) were measured by enzyme-linked immunosorbent assay (ELISA), and the LH/FSH ratio was calculated. Immunohistochemical staining (IHC) and Western blot were used to detect the protein expression levels of nucleoside binding oligomerization domain protein like receptor 3 (NALP3), apoptosis-associated speck-like protein (ASC), cysteine protease-1 (Caspase-1), nuclear transcription factor-κB (NF-κB), interleukin-1β (IL-1β), interleukin-18 (IL-18), and interleukin-6 (IL-6) in the rat ovaries. ResultAs compared with the blank group, large follicles with polycystic expansion were found in the ovaries of the model group, no dominant follicles were found, the granular layer of follicles decreased and arranged loosely, and the number of corpus luteum decreased significantly. Serum T, LH, AMH and LH/FSH increased in the model group (P<0.05, P<0.01), while FSH and E2 decreased (P<0.05, P<0.01). The relative protein expression levels of NALP3, ASC, Caspase-1, NF-κB, IL-1β, IL-18, and IL-6 increased (P<0.05, P<0.01) in the ovaries of the model group. Compared with the model group, the low, medium, and high-dose Jianpi Yishen Huazhuo prescription groups and the metformin group showed growing follicles and corpus luteum at all levels, the number of cystic expanding follicles decreased, the thickness of follicular granular layer increased, the number of follicular fluid increased, mature follicles were visible, and the local morphology of oocytes was complete. Serum T, LH, AMH, and LH/FSH in these groups decreased (P<0.05, P<0.01), while E2 and FSH increased (P<0.05). The relative protein expressions of NALP3, ASC, Caspase-1, NF-κB, IL-1β, IL-18, and IL-6 in the ovaries of these groups decreased (P<0.05, P<0.01). There was no significant difference among the treatment groups. ConclusionBy inhibiting the activation of NLRP3 inflammasome, Jianpi Yishen Huazhuo prescription reduces the release of NALP3, ASC, Caspase-1, NF-κB, IL-18, IL-1β, and IL-6 inflammatory factors in ovarian tissues, regulates endocrine level, and effectively reduces PCOS inflammatory statu, so as to play a role in improving ovarian function.
4.Research progress of cystic fibrosis associated Pseudo-Bartter syndrome
Zhiao DU ; Huiming SUN ; Zhengrong CHEN
Chinese Journal of Applied Clinical Pediatrics 2022;37(22):1727-1729
Cystic fibrosis (CF) is an autosomal recessive inherited disease affecting multiple body systems.Pseudo-Bartter syndrome (PBS) is a common manifestation of CF, with such clinical features as hypochloremia, hyponatremia, hypokalemia and metabolic alkalosis.However, PBS patients do not have renal tubulopathy.Children with CF are prone to develop electrolyte abnormalities due to fluid and electrolyte loss.In this article, the pathogenesis, clinical manifestations, diagnosis, and treatment of CF associated PBS were reviewed in order to enhance clinical understan-ding of this disease.
5.Preliminary clinical analysis of concurrent chemoradiotherapy combined with nituzumab for inoperable locally advanced esophageal cancer
Cheng ZENG ; Dan YANG ; Rongxu DU ; Leilei JIANG ; Xin DONG ; Dongming LI ; Rong YU ; Huiming YU ; Anhui SHI
Chinese Journal of Radiation Oncology 2021;30(4):353-356
Objective:The standard treatment for inoperable locally advanced esophageal cancer is concurrent chemoradiotherapy, but the survival was not satisfied. Nituzumab is a humanized IgG monoclonal antibody against EGFR. The purpose of this study is to investigate the toxicity and efficacy of concurrent chemoradiotherapy combined with nituzumab for locally advanced esophageal cancer.Methods:We retrospectively reviewed the clinical data of locally advanced esophageal cancer who were treated with concurrent chemoradiotherapy combined with nituzumab in Peking University Cancer Hospital from June 2015 to June 2020. Kaplan- Meier method was used for analysis. Results:Thirty Patients were enrolled this study.After a median follow-up of 22.5 months, The objective response rate was 93%. The 1-year, 2-year, 3-year overall survival rates were 83%, 57% and 41%, with the progression-free survival rates 75%, 47% and 32%, with the local-recurrence free survival rates 83%, 53% and 37%, with the metastasis-free survival rates 75%, 51% and 36%, respectively.The incidence of grade≥3 hematological toxicity was 32%. There were 16% patients experiencing grade≥3 esophagitis.Conclusion:The preliminary result of concurrent chemoradiotherapy combined with nituzumab is effective and safe for patients with locally advanced esophageal cancer.
6.Expert Consensus on Evaluation, Treatment and Rehabilitation of Traumatic Spinal Cord Injury
Jianjun LI ; Mingliang YANG ; Degang YANG ; Feng GAO ; Liangjie DU ; Limin LIAO ; Bohua CHEN ; Fang ZHOU ; Xuesong ZHANG ; Tiansheng SUN ; Baozhong ZHANG ; Xiaopei XIANG ; Lixia CHEN ; Hongjun ZHOU ; Songhuai LIU ; Zhihan SUN ; Ying LIU ; Xuan LIU ; Chunying HU ; Qiuchen HUANG ; Juan WU ; Fubiao HUANG ; Xiaoying ZHANG ; Jun LI ; Liang CHEN ; Hongwei LIU ; Huiming GONG
Chinese Journal of Rehabilitation Theory and Practice 2017;23(3):274-287
Spinal cord injury is a catastrophic injury causing lifelong severe disabilities, and poses a great burden to the individuals, families and society. In order to promote the standardization in treatment of traumatic spinal cord injury, the consensus on the evaluation, treatment and rehabilitation of traumatic spinal cord injury was suggested by experts, who came from authoritative multicenter in China. The expert consensus, which formed a standardization process from the first aid clinical treatment to rehabilitation of spinal cord injury, shall give a better practical guide for clinic and rehabilitation physicians.
7.Survey on Awareness of Upper Limb Function Reconstruction in Patients with Cervical Spinal Cord Injury
Jun LI ; Chong WANG ; Liangjie DU ; Hongwei LIU ; Mingliang YANG ; Feng GAO ; Yun GUO ; Liang CHEN ; Degang YANG ; Huiming GONG ; Jianjun LI
Chinese Journal of Rehabilitation Theory and Practice 2017;23(2):217-220
Objective To survey the attitudes and acceptance level of upper limb function reconstruction in patients with cervical spinal cord injury and analyze the related factors. Methods From October, 2013 to June, 2014, a total of 104 patients with cervical spinal cord injury were surveyed with self-designed questionnaire. Results There were 81 valid questionnaires, in which 3 patients (3.7%) heard of upper-extremity reconstructive surgery. The patients longed for upper limb function improvement most. After they had a understanding of the surgery, their willingness improved (P<0.01). No significant difference was found in the satisfaction degree and operation willingness between different hand function groups (P>0.05). The patients focused on the operation cost, the operation risk, recovery time after operation and the satisfaction of assistive devices, however, they were not correlated with the willingness of upper limb function reconstruction (P>0.05). Conclusion There is a clear gap between strong desire for functional improvement of upper limb and surgery awareness. It is important to communicate upper limb among patients with cervical spinal cord injury.
8.Effect of dexmedetomidine on synaptic transmission in spinal dorsal horn of rats
Shibin DU ; Jing LI ; Xiaoqin LI ; Huiming LI ; Yan LYU ; Hongjie YUAN
Chinese Journal of Anesthesiology 2016;36(10):1232-1235
Objective To evaluate the effect of dexmedetomidine on synaptic transmission in the spinal dorsal horn of rats.Methods Male Sprague-Dawley rats,aged 4-6 weeks,weighing 150-200 g,were used in the study.The lumbar enlargemnent segments of the spinal cord were harvested,and the parasagittal lumbosacral spinal cord slices with attached dorsal roots were prepared and incubated in artificial cerebro-spinal fluid.The whole-cell patch-clamp technique was used to record each index,and 4 spinal cord slices were selected and used for each index records.Experiment Ⅰ Dexmedetomidine was added cumulatively in concentration increments.Aδ and C fibers-mediated evoked excitatory postsynaptic currents (eEPSCs) were recorded before administration (baseline) and during perfusion with dexmedetomidine 4 and 10 μg/ml.Experiment Ⅱ The neurons innervated by Aδ and C fibers were selected,and Aδ and C fibers-mediated eEPSCs were recorded before administration (baseline),at 5 min of perfusion with yohimbine (alpha 2 adrenergic receptor antagonist) 2 μmol/L,and during continuous perfusion with yohimbine 2 μmol/L plus dexmedetomidine 4 μg/ml.Experiment Ⅲ The evoked excitatory postsynaptic potentials (eE-PSPs) and evoked inhibitory postsynaptic potentials (eIPSPs) were recorded before administration (baseline) and during perfusion with dexmedetomidine 4 μg/ml.Results Dexmedetomidine could dose-dependently inhibit Aδ and C fibers-mediated eEPSCs,dexmedetomidine could inhibit Aδ and C fibers-mediated eEPSPs and produced no effect on eIPSPs,and yohimbine could inhibit dexmedetomidine-induced inhibitory effect on eEPSCs.Conclusion The mechanism by which dexmedetomidine inhibits nociceptive information transmission in the spinal dorsal horn is related to inhibition of excitatory synaptic transmission through activating α2-adrenergic receptors,but not related to activation of inhibitory synaptic transmission in rats.
9.Prevention and Treatment Effect of Yiqi Huoxue Fang on Deep Venous Thrombosis
Li ZHANG ; Feng QIN ; Huiming ZHANG ; Jiehui LI ; Chunxia ZHANG ; Juanjiao DU
Herald of Medicine 2016;35(8):823-827
Objective To explore the intervention mechanism of Yiqi Huoxue Fang on deep venous thrombosis ( DVT) of rabbit model,by observing its effects on serum t-PA and PAI-1,D-D,NO,CRP,PT,blood rheology and vascular pathology. Methods Fifty New Zealand rabbits were randomly selected.Ten rabbits were served as blank control group.DVT animal models were established in the remaining 40 rabbits, which were randomly divided into model control group, Yiqi Fang group, Huoxue Fang group and Yiqi Huoxue Fang group (n=10 each group).The changes of serum t-PA,PAI-1,D-D,NO,CRP,PT and pathological changes of blood vessel were observed after 7-days treatment. Results As compared with the blank control group, the serum t-PA,PAI-1,D-D,CRP and PT were significantly increased in the model control group,Yiqi Fang group,Huoxue Fang group and Yiqi Huoxue Fang group (all P<0.01); NO was decreased (P<0.01 or P<0.05 in model control group,Yiqi Fang group and Huoxue Fang group;P>0.05 in Yiqi Huoxue Fang group vs.the blank control group).As compared with model control group,the serum t-PA,NO,D-D increased,PAI-1 and CRP value reduced in Yiqi Huoxue Fang group and Huoxue fang group (P<0.01 or P<0.05).As compared with Yiqi Huoxue Fang group,serum t-PA in both Yiqi Fang group and Huoxue Fang group was significantly lower (P<0.01 and P<0.05) ,other indexes were significantly higher than that in Yiqi Huoxue Fang group (serum PAI-1,P<0.01 in Yiqi Fang group;NO and D-D,P<0.01 or P<0.05 in Yiqi Fang group;CRP value,P<0.01).Vascular pathology showed that blood vessels,especially vascular endothelia,of Yiqi Huoxue Fang group were most slightly injured,with slightest infiltration of inflammatory cells. Conclusion Yiqi Huoxue Fang can prevent DVT through regulating t-PA and PAI-1,keeping the fibrinolytic system function at a relatively high level,protecting vascular intima and relieving the inflammation in venous thrombosis.
10.Changes of Serum S-100βProtein after Complete Spinal Cord Injury
Yuming WANG ; Zhimin ZHU ; Junyan CHE ; Huilan LI ; Ke MA ; Huiming GONG ; Hui CHEN ; Hua ZHONG ; Mingliang YANG ; Liangjie DU
Chinese Journal of Rehabilitation Theory and Practice 2016;22(7):824-826
Objective To evaluate whether S-100β protein could be a serum marker for traumatic spinal cord injury (SCI). Methods From June, 2013 to October, 2014, 24 patients with complete SCI were measured the serum S-100β protein concentrations with en-zyme-linked immunosorbent assay, one week, three and six months after SCI. Serum from ten healthy persons was as normal control. Re-sults The serum S-100βprotein concentrations increased one week and 3 months after SCI (Z>4.273, P<0.001). Conclusion The increase of serum S-100βprotein may help assessing early impairment after complete SCI.


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