OBJECTIVE To study the pharmacological substance basis of Shaoyao gancao decoction for relieving acute and chronic pain. METHODS The antispasmodic effect of Shaoyao gancao decoction， ethyl acetate extract of Shaoyao gancao decoction and its effluent part of macroporous resin and 90% ethanol elution part of macroporous resin （the concentration of 4 drugs was 13.44 g/mL according to crude drug） was observed by in vitro small intestine tension test in rats. The acetic acid writhing test was conducted in mice to evaluate the analgesic effects of macroporous resin efflux site and macroporous resin 90% ethanol elution site （the dosage of 2.4 g/kg according to crude drug）. The levels of tumor necrosis factor-α （TNF-α）， interleukin-1β （IL- 1β）， prostaglandin E2 （PGE2） and cyclooxygenase-2 （COX-2） in serum of mice were detected. The serum prototype and metabolites of mice after intragastric administration of macroporous resin 90% ethanol elution site were identified by high performance liquid chromatogre-time-of-flight mass spectrometry. RESULTS In vitro experiment showed that 90% ethanol eluting part of macroporous resin represented the best antispasmodic effect， and the inhibitory rate of small intestine tension was significantly higher than macroporous resin efflux site of Shaoyao gancao decoction （P＜0.05） without statistical significance， compared with Shaoyao gancao decoction （P＞0.05）. In the acetic acid writhing experiment， compared with model group， the writhing times of mice in the macroporous resin 90% ethanol elution part group were reduced significantly （P＜0.05）， the writhing latency was prolonged significantly （P＜0.05）， and the levels of COX-2， IL-1β， PGE2 and TNF-α in serum were decreased significantly （P＜0.05）. Ten kinds of protoproducts including paeoniflorin and glycyrrhizic acid were identified from serum of mice， and twenty-two kinds of metabolites including hydroxylated glycyrrhizin and glucosylated liquiritin were identified. CONCLUSIONS The effective part of Shaoyao gancao decoction for relieving acute and chronic pain is 90% ethanol elution part prepared by macroporous resin from the ethyl acetate extract. Ten components， including glycyrrhetinic acid and paeoniflorin， may be the basis of its pharmacological substances.

【Objective】 To analyze the height growth of children aged 6 - 15 years in Nanning, Guangxi Zhuang Autonomous Region, so as to provide evidence for the assessment of local children′s height development. 【Methods】 A total of 25 225 children aged 6 - 15 years were selected to get their physical examination data from 41 primary schools in Nanning by stratified cluster sampling method in December 2021.Then the height data were compared with the current domestic standards. 【Results】 The average height of boys in Nanning was lower than the national standard before the age of 10 years and 7 months, and the gap with the national standard gradually narrowed after the age of 10 years and 7 months. The average height of boys in Nanning City exceeded the national standard between the age of 11 years and 1 month and 13 years and 6 months, and then lagged behind the national standard again after the age of 13 years and 7 months. The mean height of girls in Nanning City was lower than the national standard height in several age groups, and it was more obvious before the age of 9 years and 7 months. The proportion of height ≤-2s,≤-s,≥ +s and ≥+2s in boys aged 6 to 15 years in Nanning City fluctuated from 2.59% to 6.04%, 12.09% to 23.43%, 7.18% to 18.79% and 0.93% to 3.14%, respectively; the total proportions were 4.56%, 17.46%, 11.35% and 1.74%, respectively; the minimum/maximum proportion values of each height group were at 11 years old /8 years old, 14 years old/8 years old, 8 years old/12 years old, and 6 years old/11 years old, respectively. The proportions of girls aged 6 - 15 years in Nanning City with height ≤-2s, ≤-s, ≥+s, and ≥+2s fluctuated from 2.06% to 5.19%, 9.35% to 25.15%, 8.21% to 15.80% and 1.23% to 3.49%, respectively; the total proportions were 3.38%, 16.91%, 11.97% and 2.29%, respectively; and the minimum/maximum proportion values of each height group were at 13 years old/6 years old, 12 years old/6 years old, 7 years old/12 years old, and 6 years old/11 years old, respectively. 【Conclusions】 The overall height level of children in Nanning is still lower than the national level, with short prepubertal basal heights, an earlier age of onset of accelerated pubertal height, and a shorter duration of accelerated pubertal height in boys. Strengthening pre-pubertal height management and emphasizing the onset and duration of children′s pubertal development, especially the height development of boys during puberty, can help improve the adult lifelong height of children in this region.

Endocrine-resistance remains a major challenge in estrogen receptor α positive (ERα⁺) breast cancer (BC) treatment and constitutively active somatic mutations in ERα are a common mechanism. There is an urgent need to develop novel drugs with new mode of mechanism to fight endocrine-resistance. Given aberrant ERα activity, we herein report the identification of novel covalent selective estrogen receptor degraders (cSERDs) possessing the advantages of both covalent and degradation strategies. A highly potent cSERD 29c was identified with superior anti-proliferative activity than fulvestrant against a panel of ERα⁺ breast cancer cell lines including mutant ERα. Crystal structure of ERα‒ 29c complex alongside intact mass spectrometry revealed that 29c disrupted ERα protein homeostasis through covalent targeting C530 and strong hydrophobic interaction collied on H11, thus enforcing a unique antagonist conformation and driving the ERα degradation. These significant effects of the cSERD on ERα homeostasis, unlike typical ERα degraders that occur directly via long side chains perturbing the morphology of H12, demonstrating a distinct mechanism of action (MoA). In vivo, 29c showed potent antitumor activity in MCF-7 tumor xenograft models and low toxicity. This proof-of-principle study verifies that novel cSERDs offering new opportunities for the development of innovative therapies for endocrine-resistant BC.

Objective:To determine the effects of menopausal stage, age and other associated risk factors on symptoms of anxiety and depression among women in a community in Beijing.Methods:This study was a community-based prospective cohort. Participants who had transitioned through natural menopause, completed two or more depressive and anxiety symptoms evaluations, aged 35 to 64 years, and did not use hormone therapy were selected from the Peking Union Medical College Hospital aging longitudinal cohort of women in midlife to this analysis. The primary outcome variables were depressive and anxiety symptoms, assessed by hospital anxiety and depression scale (HADS). The generalized estimation equation was used in the statistical analysis.Results:Followed up from 2006 to 2014, 430 women and 2 533 HADS assessments were retained in the cohort. Depressive symptoms were more common than anxiety symptoms during all menopausal stages. The incidences of depressive and anxiety symptoms were 14.5% (19/191) and 3.1% (4/191) in the premenopausal -3 stage, respectively. The incidence increased in both menopausal transition and postmenopausal stage, with the highest incidence in the +1c stage [20.6% (155/751) and 8.8% (66/751), respectively]. However, these differences were not statistically significant (all P>0.05). Depressive symptoms were highest in the ≥60-<65 age group [20.8% (74/355)], and anxiety symptoms were highest in the ≥50-<55 age group [8.2% (62/754)]; but there were no statistical significances between different age groups and depressive and anxiety symptoms (all P>0.05). Multivariable analysis showed that high body mass index, low education status, and poor health status were independently associated with depressive symptoms (all P<0.05), and that poor health status, trouble falling asleep, and early awakening were independently associated with anxiety symptoms (all P<0.01). Conclusions:Depressive and anxiety symptoms are more common during menopausal transition and postmenopausal stage compared with reproductive stage. Depressive symptoms are more common than anxiety symptoms. To screen and assess depressive and anxiety symptoms in perimenopausal women is essential, especially for women with high risk factors.

Objective:To evaluate the role of extracellular signal-regulated kinase 1/2 (ERK1/2)/cyclic adenosine monophosphate response element-binding protein (CREB)/brain-derived neurotrophic factor (BDNF) signaling pathway in dexmedetomidine-induced inhibition of propofol-caused apoptosis in isolated hippocampal neurons of fetal rats.Methods:Pregnant Sprague-Dawley rats at 16 days of gestation were sacrificed, and the fetal rats were taken out, and hippocampal neurons of fetal rats were obtained and primarily cultured in vitro for 7 days.The neurons were divided into 9 groups ( n=12 each) using a random number table method: control group (group C), fat emulsion group (group I), dimethyl sulfoxide (DMSO) group, dexmedetomidine group (group D), propofol group (group P), propofol plus dexmedetomidine group (group PD), PD98059 plus propofol plus dexmedetomidine group (group PDP), MH89 plus propofol plus dexmedetomidine group (group HDP), and KG501 plus propofol plus dexmedetomidine group (group KDP). Group C received no treatment.In group I, 20% fat emulsion was added, and the neurons were incubated for 30 min, and 0.25% DMSO was added in group DMSO, and the neurons were incubated for 30 min.Dexmedetomidine at a final concentration of 10 μmol/L was added, and the neurons were incubated for 30 min in group D. Propofol at a final concentration of 100 μmol/L was added, and the neurons were incubated for 3 h in group P. In group PD, dexmedetomidine at a final concentration of 10 μmol/L was added, the neurons were incubated for 30 min, propofol at a final concentration of 100 μmol/L was added, and the neurons were incubated for 3 h. In PDP, HDP and KDP groups, 25 μmol PD98059 (p-ERK1/2 inhibitor), 10 μmol H89 (p-CREB inhibitor) and 25 μmol KG501 (CREB inhibitor) were added, respectively, the neurons were incubated for 30 min, dexmedetomidine at a final concentration of 10 μmol/L was added, the neurons were incubated for 30 min, and propofol at a final concentration of 100 μmol/L was added, and the neurons were incubated for 3 h. The cell ultrastructure was observed with the transmission electron microscope, the apoptosis in neurons was detected by flow cytometry, the expression of ERK1/2, CREB and BDNF mRNA was detected by quantitative real-time polymerase chain reaction, and the expression of p-ERK1/2, CREB, p-CREB, BDNF and cleaved caspase-3 was detected by Western blot. Results:Compared with group C, the apoptosis rate was significantly increased, the expression of p-ERK1/2 and p-CREB was down-regulated, and the expression of cleaved caspase-3 was up-regulated in P, PD, PDP, HDP and KDP groups, and the expression of BDNF was significantly down-regulated in P, PDP, HDP and KDP groups ( P<0.05). Compared with group P, the apoptosis rate was significantly decreased, the expression of p-ERK1/2, p-CREB and BDNF was up-regulated, and the expression of cleaved caspase-3 was down-regulated in group PD ( P<0.05). Compared with group PD, the apoptosis rate was significantly increased, the expression of p-ERK1/2, p-CREB and BDNF was down-regulated, and the expression of cleaved caspase-3 was up-regulated in PDP, HDP and KDP groups ( P<0.05). Conclusion:The ERK1/2/CREB/BDNF signaling pathway is involved in dexmedetomidine-induced inhibition of propofol-caused apoptosis in isolated hippocampal neurons of fetal rats.

Objective To evaluate the role of cyclic adenosine monophosphate-protein kinase A-cAMP response element-binding protein (cAMP-PKA-CREB) signaling pathway in hypoxic preconditioninginduced reduction of propofol-induced central neurotoxicity in the developing rats.Methods A total of 70 SPF male Sprague-Dawley rats,aged 7 days,weighing 10-15 g,were divided into 7 groups (n=10 each) using a random number table method:normal saline group (N group),propofol group (P group),hypoxic preconditioning plus propofol group (HP group),hypoxic preconditioning plus propofol plus PKA inhibitor H89 group (HPH group),propofol plus PKA agonist SP-CAMP group (PS group),normal saline injected via the lateral cerebral ventricle group (NI group),and 5％ dimethyl sulfoxide (DMSO) injected via the lateral cerebral ventricle group (DI group).In P group,propofol 50 mg/kg was intraperitoneally injected,and an increment of propofol 50 mg/kg was given after recovery of righting reflex.The equal volume of normal saline was given instead in N group.In HP group,hypoxic preconditioning (rats were subjected to 5 cycles of 10-min hypoxia of 8％ O2 and 1O-min normoxia of 21％ O2) was performed,and propofol was intraperitoneally injected at 2 h after the end of hypoxic preconditioning and the method was similar to those previously described in P group.In HPH group,H89 5 μmol/5 μl was injected via the lateral cerebral ventricle,and 30 min later the other treatment was similar to those previously described in HP group.In PS group,SP-CAMP 20 nmol/5 μl was injected via the lateral cerebral ventricle,and 30 min later propofol was injected using the method previously described in P group.In NI and DI groups,5 μl normal saline and 5％ DMSO were injected via the lateral cerebral ventricle,respectively.Rats were immediately sacrificed after the righting reflex was recovered,brains were removed and hippocampi were isolated and cut into sections which were stained with haematoxylin and eosin for determination of PKAc and p-CREB positive cells (by i mmuno-histochemistry) and expression of cleaved caspase-3,Bcl-2,Bax,PKAc and posphorylated (p-CREB) protein (by Western blot).Results Compared with N group,the expression of cleaved caspase-3 and Bax was significantly up-regulated,the expression of Bcl-2,PKAc and p-CREB was downregulated,and the percentage of PKAc and p-CREB positive cells was decreased (P＜0.05),hippocampal cells had irregular arrangement,and cells was atrophied in P group.Compared with P group,the expression of cleaved caspase-3 was significantly down-regulated,the expression of Bcl-2,PKAc and p-CREB was up-regulated,and the percentage of PKAc and p-CREB positive cells was increased in HP and PS groups,and the expression of Bax was down-regulated (P＜0.05),the hippocampal cells were arranged neatly,the cytoplasm was abundant,and the nuclei were visible in HP group.Compared with HP group,the expression of cleaved caspase-3 was significantly up-regulated,the expression of Bcl-2,PKAc and p-CREB protein was down-regulated and the percentage of PKAc and p-CREB positive cells was decreased (P＜0.05),the cells had irregular arrangement and shrinked,and nuclear condensation was found in cells in HPH group.Conclusion The mechanism by which hypoxic preconditioning reduces propofol-induced central neurotoxicity may be related to activating cAMP-PKA-CREB signaling pathway in the developing rats.

Objective To explore the long-term effects of dexmedetomidine on the brain development in propofol-induced neonatal rats.Methods Thirty-five seven-day-old Sprague-Dawley rats of both genders,weighing 10-15 g,were randomly divided into seven groups (n =5) using a random number table:normal saline group (group N),intralipid group (group F),propofol 100 mg/kg group (group P),dexmedetomidine 75 μg/kg (group D),dexmedetomidine 25 μg/kg,50 μg/kg and 75μg/kg+propofol 100 mg/kg groups (groups PD25,PD50 and PD75),neonatal rats in each group were treated according to the corresponding dosing regimen.After fully awake,the rats were allowed to mature until postnatal week 9 and the spatial learning and memory capacities were tested by Morris water maze.The rats were sacrificed after the tests.Brain was sliced for determination of hippocampal apoptosis by TUNEL assays and the expression of postsynaptic density protein 95 (PSD95) by immunohistochemistry.Results Compared with group N,the escape latency was significantly prolonged,the times of platform crossing were significantly decreased,the hippocampal apoptosis ratio was significantly increased and the expression of PSD95 was significantly down-regulated in groups P,PD25 and PD50 (P＜0.05).Compared with group P,the escape latency was significantly shortened,the times of platform crossing were significantly increased and the hippocampal apoptosis were significantly decreased in groups PD50 and PD75 (P＜0.05),the expression of PSD95 was up-regulated in group PD75 (P＜0.05).Compared with group PD25,the escape latency was significantly shortened,the number of platform crossing was significantly increased and the hippocampal apoptosis were significantly decreased in groups PD50 and PD75 (P＜0.05),the expression of PSD95 was significantly up-regulated in group PD75 (P＜0.05).Compared with group PD50,the hippocampal apoptosis were significantly decreased,the expression of PSD95 was significantly up-regulated in group PD75 (P＜ 0.05).Conclusion The addition of dexmedetomidine 50,75 μg/kg attenuates propofol-induced neurocognitive impairment in neonatal rats after aducthood,partially by attenuating hippocampal apoptosis and upregulating the expression of PSD95.Dexmedetomidine alone was not neurotoxic to the developing brain.

Objective To evaluate the role of hippocampal phosphatidylinositol 3-kinase/serinethreonine kinase/glycogen synthase kinase-3 beta (PI3K/Akt/GSK-3β) signaling pathway in dexmedetomidine-induced reduction of long-term cognitive decline caused by propofol in neonatal rats.Methods A total of 110 clean healthy male Sprague-Dawley rats,aged 7 days,weighing 10-15 g,were divided into 11 groups (n=10 each) using a random number table:normal saline group (NS group),fat emulsion group (F group),10％ dimethyl sulfoxide (DMSO) group (D2 group),dexmedetomidine group (DEX group),TDZD-8 group (TDZD group),10％ DMSO group (D1 group),LY294002 group (LY group),propofol group (P group),dexmedetomidine plus propofol group (PD group),LY294002 plus dexmedetomidine plus propofol group (LYPD group) and TDZD-8 plus dexmedetomidine plus propofol group (TDPD group).Normal saline,fat emulsion,10％ DMSO 100 μl,dexmedetomidine 75 μg/kg and TDZD-8 1 mg/kg were intraperitoneally injected in NS,F,D2,DEX and TDZD groups,respectively.10％ DMSO 5 μ1 and LY294002 25 μg/5 μl were injected via the lateral cerebral ventricle in D1 and LY groups,respectively.Propofol 50 mg/kg was intraperitoneally injected,and an increment of propofol 50 mg/kg was given after recovery of righting reflex in group P.Dexmedetomidine 75 μg/kg was intraperitoneally injected and 30 min later propofol was injected in group PD.LY294002 was injected,30 min later dexmedetomidine was injected,and 30 min later propofol was injected in group LYPD.In group TDPD,TDZD-8 was injected and the other treatment was similar to those previously described in group LYPD.Then the rats were fed to 9 weeks old after returning to the state of consciousness.Morris water maze test was performed to evaluate the cognitive function.Rats were then sacrificed and their hippocampi were harvested for detection of the expression of PI3K,Akt,GSK-3β and PSD95 mRNA (using real-time polymerase chain reaction) and expression of Akt,pAkt(ser473),GSK-3β,pGSK-3β(ser9) and PSD95 (by Western blot).Results Compared with NS group,the escape latency was significantly prolonged,the number of crossing the original platform was reduced,the expression of PI3K,Akt and PSD95 mRNA was down-regulated,the expression of GSK-3β mRNA was up-regulated,p-Akt(ser473)/Akt ratio was decreased,the expression of PSD95 was downregulated,and pGSK-3β (ser9)/GSK-3β ratio was increased in P group (P＜ 0.05).Compared with P group,the escape latency was significantly shortened and the number of crossing the original platform was increased in PD,LYPD and TDPD groups,the expression of PI3K,Akt and PSD95 mRNA was up-regulated,and the expression of GSK-3β mRNA was down-regulated in PD group,and pAkt(ser473)/Akt ratio was increased,the expression of PSD95 was up-regulated,and pGSK-3β (ser9)/GSK-3β ratio was decreased in PD and TDPD groups(P＜0.05).Compared with PD group,the escape latency was significantly prolonged,the number of crossing the original platform was reduced,the expression of GSK-3β mRNA was up-regulated,the expression of PSD95 mRNA was down-regulated,pAkt (ser473)/Akt ratio was decreased,the expression of PSD95 was down-regulated,and pGSK-3β (ser9)/GSK-3β ratio was increased in LYPD group,and the escape latency was significantly shortened,the number of crossing the original platform was increased,the expression of GSK-3β mRNA was down-regulated,the expression of PSD95 mRNA was up-regulated,pGSK-3β(ser9)/GSK-3β ratio was decreased,and the expression of PSD95 was up-regulated in TDPD group(P＜0.05).Conclusion Hippocampal PI3K/Akt/GSK-3β signaling pathway is involved in dexmedetomidine-induced reduction of long-term cognitive decline caused by propofol in neonatal rats.

Objective To investigate the effects of intrathecal injection of Roscovitine on the pain behavior and the expression of pERK1/2 and pPPARγ receptor in spinal cord in rats with neuropathic pain.Methods Fifty-two male adult Sprague-Dawley rats of 220～ 250 g were randomly divided into 4 groups:Sham+DMSO group (dimethyl sulfoxide,S+D group),Sham+Roscovitine (S+R) group,CCI+ DMSO group (I+D),CCI+ Roscovitine (I+R) group.The corresponding drugs were administered by intrathecal injection from the seventh day after CCI once a day for 14 days.The right hind paw mechanical threshold value (PMWT) was measured by Von Frey filament and paw thermal withdrawal latency(PWTL) was measured by Hargreaves methods before 1 day and 3 d,7 d,10 d,14 d after surgery.The spinal cord lumbar enlargement was taken at 14 d after surgery,and Cdk5,p35,phosphorylated ERK protein (pERK),total ERK protein (ERK),phosphorylated PPARγprotein (pPPARγ) and total PPARγ protein (PPARγ) were detecteded by Western blot.Results Roscovitine was administered by intrathecal injection alleviated mechanical allodynia and thermal hyperalgesia of CCI rats.Compared with I+D group,PWMT in I+R group at 7 d,10 d,14 d after administration((4.65±0.08)g,(6.47±0.12)g,(7.90±0.19)g,) vs ((3.71 ±0.06)g,(2.45±0.17)g,(2.31±0.15)g) (Fgroup =505.71,P＜0.05,Ftime×group =15.33,P＜0.05),PWTL((9.22±0.33) s,(13.52±0.43) s,(12.63±0.88) s) vs ((8.02±0.20) s,(5.90±0.28) s,(5.40±0.38) s) (Fgroup =355,P＜0.05,F time×group =8.589,P＜0.05) were significantly increased.Compared with I+D group(p35 (0.58±0.02),pERK (1.12±0.13),pPPARγ (0.77±0.16)),p35 (0.46±0.04,F=11.06,P＜0.05) and pERK(0.79±0.11,F =22.91,P＜ 0.05) in I+ R group,were significantly dereased,and the expression of pPPARγ(0.99±0.13,F =17.62,P＜0.05))was significantly increased.Conclusion Intrathecal injection Roscovitine can alleviate both mechanical allodynia and thermal hyperalgesia in rats with neuropathic pain,and its mechanisms may be related to the inhibition of Cdk5/p35 and ERK activity,enhancement of PPARγ activity in the spinal dorsal horn.

Objective To explore the effect of dexmedetomidine on phosphoinositide 3-kinase/protein kinase B (PI3K/Akt ) pathway in hippocampus of propofol anesthetized neonatal rats. Methods Eighty Sprague-Dawley male rats,aged 7 days,weighing 10-1 5 g,were randomly divided into 8 groups (n= 10 each):normal saline group (group N),DMSO group (group D),intralipid group (group I),propofol group (group P),dexmedetomidine 25 μg/kg,50 μg/kg and 75 μg/kg +propofol 100 mg/kg groups (groups PD25 ,PD50 and PD7 5 ),LY294002 25 μg + dexmedetomidine 75μg/kg + propofol 100 mg/kg group (group LYPD).The hippocampus of rats in all groups were taken 2 h after the animals fully awake.The ultrastructure of hippocampal neurons was observed by transmission electron microscope.The pAkt-(ser473 )protein and Akt protein in the hippocampus were evaluated by Western blot analysis.Results There was no significant difference in the expression of Akt protein among the eight groups.Compared with group N,the expression of pAkt (ser473)protein was significantly down-regulated in groups P,PD25 ,PD50 ,PD7 5 and LYPD (P <0.05).Compared with group P,the expression of pAkt (ser473)protein was increased significantly in groups PD7 5 and LYPD (P <0.05).Compared with group PD7 5 ,the expression of pAkt (ser473) protein was significantly down-regulated in group LYPD (P <0.05 ).The structure of hippocampal neurons was normal in groups N,I and D.Nuclear nuclei swelling,chromatin decreasing and mito-chondrion vacuolar degeneration were observed in group P while improved gradually with dexmedeto-midine in a dose-dependent manner in groups PD25 ,PD50 and PD7 5 .Neurons karyopyknosis,partial dissolution of nuclear membrane,chromatin condensation,mitochondria vacuolar degeneration were observed in group LYPD.Conclusion Dexmedetomidine pretreatment provides neuroprotection against propofol-induced hippocampal destruction by preserving PI3K/Akt pathway activity in the de-veloping brains.