Objective:To explore the mechanism by which moxibustion improves cold-induced blood stasis pattern(CIBSP)and to uncover the biological basis underlying the correlation between the Conception Vessel(CV),Governor Vessel(GV),Thoroughfare Vessel(TV),and the uterus by investigating the effects of mild moxibustion on the hypothalamic-pituitary-ovary axis(HPOA)in CIBSP model rats.Methods:Forty-eight female Sprague-Dawley rats were divided into a blank group,a model group,a CV group,a GV group,a TV group,and a non-meridian non-point(NMNP)group according to the random number table method,with 8 rats in each group.Except for rats in the blank group,the other rats were subjected to an ice water bath plus epinephrine to prepare CIBSP models.Each intervention group was given mild moxibustion on the first day of modeling.Qihai(CV6),Guanyuan(CV4),and Zhongji(CV3)were selected in the CV group;Huangshu(KI16),Dahe(KI12),and Henggu(KI11)were selected in the TV group;Xuanshu(GV5),Mingmen(GV4),and Yaoyangguan(GV3)were selected in the GV group;three NMNP spots were selected in the NMNP group.Mild moxibustion was administered for 20 min each time,once a day,for 14 consecutive days.The symptom scores of CIBSP and uterine electromyogram were compared among groups;morphological changes in reproductive organs were observed by hematoxylin-eosin staining;enzyme-linked immunosorbent assay was used to detect serum reproductive hormones and neurotransmitters.The mRNA and protein expression levels of estrogen receptor(ER),follicle-stimulating hormone receptor(FSHR),and luteinizing hormone receptor(LHR)were detected by real-time quantitative polymerase chain reaction,immunohistochemistry,or Western blotting assays.Results:Compared to the blank group,the model group showed a significantly increased CIBSP symptom score,an amplified in vivo uterine myoelectric value;decreased ovarian follicles,thinner endometrial epithelium,and disordered structure with the pathological examination,decreased serum levels of reproductive hormones,6-keto-prostaglandin F1α(6-keto-PGF1α),and β-endorphin(β-EP),increased thromboxane B2(TXB2),5-hydroxytryptamine(5-HT),and substance P(SP),and decreased mRNA and protein expression of ovarian ER,FSHR,and LHR(P<0.05 or P<0.01).Compared to the model and NMNP groups,each point-intervention group showed a decreased CIBSP symptom score and uterine myoelectric value;increased ovarian follicles,thickened endometrium,increased glands,up-regulated serum reproductive hormones,6-keto-PGF1α,and β-EP,down-regulated serum TXB2,5-HT,and SP and increased mRNA and protein expression of ovarian ER,FSHR,and LHR(P<0.05 or P<0.01).Conclusion:Moxibustion at points from the CV,GV,and TV("three branches from one source")can improve the reproductive neuroendocrine function of CIBSP rats,and this effect may be mediated through the regulation of the HPOA.These detections suggest that the biological basis of point-uterus correlation may be based on HPOA regulation.

Objective To observe the effect of mild moxibustion at"Qihai","Guanyuan"and"Zhongji"on GnRH and its receptor in rats with gynecological cold coagulation and blood stasis syndrome(CCBS);To explore the mechanism of moxibustion on reproductive function of gynecological CCBS.Methods Totally 48 female SD rats were randomly divided into blank group,model group,moxibustion group,inhibitor group,inhibitor+moxibustion group and angonist group,with 8 rats in each group.The gynecological CCBS model was prepared by ice bath and adrenalin injection.GnRHant or GnRHa injection and mild moxibustion intervention were given to each treatment group.The score of CCBS symptoms and the amount of microcirculation blood perfusion in uterus were detected,the morphology of ovarian tissue and uterine tissue were observed by HE staining,serum reproductive hormones(GnRH,FSH,LH)and neurotransmitters(DA,GABA)contents were detected by ELISA,the expressions of GnRHR,FSHR and LHR mRNA and protein in ovarian and uterine tissue were detected by immunohistochemistry,RT-qPCR and Western blot.Results Compared with the blank group,the symptom score of CCBS in model group was significantly increased,the amount of microcirculation blood perfusion in uterus was decreased,the ovarian follicles were decreased,and the endometrial epithelium was thinner and the structure was chaotic,the contents of serum GnRH,FSH,LH,GABA decreased,DA content increased;the mRNA and protein expressions of GnRHR,FSHR and LHR in ovarian tissue decreased(P<0.01).Compared with the model group,the symptom score of CCBS in each intervention group were significantly decreased,the amount of microcirculation blood perfusion in uterus increased,ovarian follicles increased,endometrium thickened and glands increased,the contents of serum GnRH,FSH,LH,GABA increased,DA content decreased;the mRNA and protein expressions of GnRHR,FSHR and LHR were increased(P<0.05,P<0.01).Conclusion Moxibustion can regulate GnRH and its receptor and improve HPO axis function by regulating neurotransmitters,so as to improve the general state and reproductive function of gynecological CCBS model rats.

Objective:To explore the mechanism by which moxibustion improves cold-induced blood stasis pattern(CIBSP)and to uncover the biological basis underlying the correlation between the Conception Vessel(CV),Governor Vessel(GV),Thoroughfare Vessel(TV),and the uterus by investigating the effects of mild moxibustion on the hypothalamic-pituitary-ovary axis(HPOA)in CIBSP model rats.Methods:Forty-eight female Sprague-Dawley rats were divided into a blank group,a model group,a CV group,a GV group,a TV group,and a non-meridian non-point(NMNP)group according to the random number table method,with 8 rats in each group.Except for rats in the blank group,the other rats were subjected to an ice water bath plus epinephrine to prepare CIBSP models.Each intervention group was given mild moxibustion on the first day of modeling.Qihai(CV6),Guanyuan(CV4),and Zhongji(CV3)were selected in the CV group;Huangshu(KI16),Dahe(KI12),and Henggu(KI11)were selected in the TV group;Xuanshu(GV5),Mingmen(GV4),and Yaoyangguan(GV3)were selected in the GV group;three NMNP spots were selected in the NMNP group.Mild moxibustion was administered for 20 min each time,once a day,for 14 consecutive days.The symptom scores of CIBSP and uterine electromyogram were compared among groups;morphological changes in reproductive organs were observed by hematoxylin-eosin staining;enzyme-linked immunosorbent assay was used to detect serum reproductive hormones and neurotransmitters.The mRNA and protein expression levels of estrogen receptor(ER),follicle-stimulating hormone receptor(FSHR),and luteinizing hormone receptor(LHR)were detected by real-time quantitative polymerase chain reaction,immunohistochemistry,or Western blotting assays.Results:Compared to the blank group,the model group showed a significantly increased CIBSP symptom score,an amplified in vivo uterine myoelectric value;decreased ovarian follicles,thinner endometrial epithelium,and disordered structure with the pathological examination,decreased serum levels of reproductive hormones,6-keto-prostaglandin F1α(6-keto-PGF1α),and β-endorphin(β-EP),increased thromboxane B2(TXB2),5-hydroxytryptamine(5-HT),and substance P(SP),and decreased mRNA and protein expression of ovarian ER,FSHR,and LHR(P<0.05 or P<0.01).Compared to the model and NMNP groups,each point-intervention group showed a decreased CIBSP symptom score and uterine myoelectric value;increased ovarian follicles,thickened endometrium,increased glands,up-regulated serum reproductive hormones,6-keto-PGF1α,and β-EP,down-regulated serum TXB2,5-HT,and SP and increased mRNA and protein expression of ovarian ER,FSHR,and LHR(P<0.05 or P<0.01).Conclusion:Moxibustion at points from the CV,GV,and TV("three branches from one source")can improve the reproductive neuroendocrine function of CIBSP rats,and this effect may be mediated through the regulation of the HPOA.These detections suggest that the biological basis of point-uterus correlation may be based on HPOA regulation.

BACKGROUND:Previous studies have found that for postmenopausal osteoporosis patients,the Bushen Jianpi Formula can help regulate serum osteoclast-and osteoblast-related factors and intestinal flora levels,improve clinical symptoms of patients and further improve efficacy.OBJECTIVE:To investigate the effect of Bushen Jianpi Formula on bone metabolism and bone microarchitecture in ovariectomized rats.METHODS:Twenty-four female SD rats were randomly divided into four groups by random number table method:sham operation group(n=6)was subjected to periovarian fat removal,and bilateral ovariectomy was performed in model group(n=6),alendronate group(n=6),and Bushen Jianpi Formula group(n=6).Five days after modeling,the Bushen Jianpi Formula group was given Bushen Jianpi Formula by intragastric administration(once a day).The alendronate sodium group was given alendronate by intragastric administration(once a week).The sham operation group and model group were given an equal volume of normal saline by intragastric administration(once a day)for 12 consecutive weeks.After intragastric administration,serum levels of procollagen Ⅰ N-terminal peptide,β-isomerized C-terminal telopeptide of type Ⅰ collagen degradation product,and tumor necrosis factor α were detected.The distal femur microstructure was detected by Micro-CT.The femoral tissue morphology was observed by hematoxylin-eosin staining.The expression of type Ⅰ collagen and tumor necrosis factorα in the femur was detected by immunohistochemical staining.RESULTS AND CONCLUSION:(1)The serum ELISA test showed that compared with the sham operation group,the level of procollagen Ⅰ N-terminal peptide in the model group decreased(P＜0.05),and the levels of degradation products of β-isomerized C-terminal telopeptide of type Ⅰ collagen and tumor necrosis factor α increased(P＜0.05).Compared with the model group,the levels of the procollagen Ⅰ N-terminal peptide in the Bushen Jianpi Formula group and the alendronate sodium group increased(P＜0.05),and the levels of β-isomerized C-terminal telopeptide of type Ⅰ collagen degradation products and tumor necrosis factor α decreased(P＜0.05).(2)Micro-CT three-dimensional reconstruction showed that compared with the sham operation group,the number of bone trabeculae in the model group was significantly reduced,the continuity was worsened,and the gaps were increased.Both alendronate sodium and Bushen Jianpi Formula had a positive improvement effect on this phenomenon.(3)Hematoxylin-eosin staining showed that compared with the sham operation group,the number of bone trabeculae in the model group was reduced and the shape became thinner,and the network structure was destroyed.Compared with the model group,the number of bone trabeculae in the Bushen Jianpi Formula group and the alendronate sodium group increased,the shape became thicker,and the reticular structure was restored.(4)Immunohistochemical staining showed that compared with the sham operation group,the expression of type Ⅰ collagen was reduced and tumor necrosis factor α expression was increased in the model group.Compared with the model group,the expression of type Ⅰ collagen increased and the expression of tumor necrosis factor α decreased in the Bushen Jianpi Formula group and the alendronate sodium group.The results show that the Bushen Jianpi Formula can effectively improve bone metabolism and bone microstructure in ovariectomized rats.

Purpose: This study aimed to evaluate the contrast-enhanced ultrasound with Sonazoid (Sonazoid-CEUS) features of hepatocellular carcinoma (HCC) in patients with non-alcoholic fatty liver disease (NAFLD). Methods: In this retrospective study, patients who underwent surgical resection and were histopathologically diagnosed with NAFLD or cirrhosis-related HCC were included. All patients received Sonazoid-CEUS examinations within 1 week prior to hepatic surgery. The enhancement patterns of HCC lesions were evaluated and compared between the two groups according to the current World Federation for Ultrasound in Medicine and Biology guidelines. Multivariate logistic regression analysis was used to assess the correlations between Sonazoid-CEUS enhancement patterns and clinicopathologic characteristics. Results: From March 2022 to April 2023, a total of 151 patients with HCC were included, comprising 72 with NAFLD-related HCC and 79 with hepatitis B virus (HBV) cirrhosis–related HCC. On Sonazoid-CEUS, more than half of the NAFLD-related HCCs exhibited relatively early and mild washout within 60 seconds (54.2%, 39/72), whereas most HBV cirrhosis–related HCCs displayed washout between 60 and 120 seconds (46.8%, 37/79) or after 120 seconds (39.2%, 31/79) (P<0.001). In the patients with NAFLD-related HCC, multivariate analysis revealed that international normalized ratio (odds ratio [OR], 0.002; 95% confidence interval [CI], 0.000 to 0.899; P=0.046) and poor tumor differentiation (OR, 21.930; 95% CI, 1.960 to 245.319; P=0.012) were significantly associated with washout occurring within 60 seconds. Conclusion Characteristic Sonazoid-CEUS features are useful for diagnosing HCC in patients with NAFLD.

Background Attention-deficit hyperactivity disorder (ADHD) is a common neurodevelopmental and behavioral disorder in children, often diagnosed during school age. The etiology of ADHD remains unclear; however, existing studies suggest that environmental factors, such as exposure to triclosan (TCS), may be associated with the occurrence of ADHD-like symptoms in offspring. Nevertheless, relevant research in China remains limited. Objective To investigate the impact of early pregnancy TCS exposure on ADHD-like symptoms in 7-year-old children. Methods This study was based on the Shanghai Birth Cohort (SBC) and included 662 mother-child pairs. TCS concentrations in early pregnancy urine samples were measured using high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). Demographic information was collected via questionnaires and medical record abstraction. ADHD-like symptoms in 7-year-old children were first assessed using the Strengths and Difficulties Questionnaire (SDQ). Further differentiation of ADHD-like symptom subtypes (inattentive and hyperactive/impulsive) was conducted using the SNAP-IV, a clinically validated ADHD screening tool. Negative binomial regression models were applied to evaluate the associations between prenatal TCS exposure and hyperactive behavior (SDQ assessment) as well as ADHD-like symptom subtypes (SNAP-IV assessment) in 7-year-old children. Results The positive rate of TCS in early pregnancy urine samples was 91.39%, with median concentrations of 0.69 μg·L⁻¹ and 0.63 μg·g⁻¹ before and after the creatinine adjustment, respectively. The modeling results indicated that prenatal TCS exposure was associated with an increased risk of hyperactive symptoms (SDQ assessment) in 7-year-old children (RR=1.04, 95%CI: 1.02, 1.06); the stratified analyses by children sex revealed similar effects for both boys (RR=1.04, 95%CI: 1.02, 1.07) and girls (RR=1.04, 95%CI: 1.01, 1.07). Further analysis of ADHD-like symptom subtypes showed that prenatal TCS exposure increased the risk of inattentive symptoms (RR=1.03, 95%CI: 1.00, 1.05); the sex-stratified analyses indicated associations between TCS exposure and inattentive symptoms (RR=1.03, 95%CI: 1.00, 1.07) as well as hyperactive/impulsive symptoms (RR=1.04, 95%CI: 1.01, 1.08) in girls. Conclusion Prenatal TCS exposure is associated with an increased risk of ADHD-like symptoms in 7-year-old children, primarily contributing to the risk of the inattention subtype. The impact is more pronounced in girls.

Purpose: This study aimed to evaluate the contrast-enhanced ultrasound with Sonazoid (Sonazoid-CEUS) features of hepatocellular carcinoma (HCC) in patients with non-alcoholic fatty liver disease (NAFLD). Methods: In this retrospective study, patients who underwent surgical resection and were histopathologically diagnosed with NAFLD or cirrhosis-related HCC were included. All patients received Sonazoid-CEUS examinations within 1 week prior to hepatic surgery. The enhancement patterns of HCC lesions were evaluated and compared between the two groups according to the current World Federation for Ultrasound in Medicine and Biology guidelines. Multivariate logistic regression analysis was used to assess the correlations between Sonazoid-CEUS enhancement patterns and clinicopathologic characteristics. Results: From March 2022 to April 2023, a total of 151 patients with HCC were included, comprising 72 with NAFLD-related HCC and 79 with hepatitis B virus (HBV) cirrhosis–related HCC. On Sonazoid-CEUS, more than half of the NAFLD-related HCCs exhibited relatively early and mild washout within 60 seconds (54.2%, 39/72), whereas most HBV cirrhosis–related HCCs displayed washout between 60 and 120 seconds (46.8%, 37/79) or after 120 seconds (39.2%, 31/79) (P<0.001). In the patients with NAFLD-related HCC, multivariate analysis revealed that international normalized ratio (odds ratio [OR], 0.002; 95% confidence interval [CI], 0.000 to 0.899; P=0.046) and poor tumor differentiation (OR, 21.930; 95% CI, 1.960 to 245.319; P=0.012) were significantly associated with washout occurring within 60 seconds. Conclusion Characteristic Sonazoid-CEUS features are useful for diagnosing HCC in patients with NAFLD.

Purpose: This study aimed to evaluate the contrast-enhanced ultrasound with Sonazoid (Sonazoid-CEUS) features of hepatocellular carcinoma (HCC) in patients with non-alcoholic fatty liver disease (NAFLD). Methods: In this retrospective study, patients who underwent surgical resection and were histopathologically diagnosed with NAFLD or cirrhosis-related HCC were included. All patients received Sonazoid-CEUS examinations within 1 week prior to hepatic surgery. The enhancement patterns of HCC lesions were evaluated and compared between the two groups according to the current World Federation for Ultrasound in Medicine and Biology guidelines. Multivariate logistic regression analysis was used to assess the correlations between Sonazoid-CEUS enhancement patterns and clinicopathologic characteristics. Results: From March 2022 to April 2023, a total of 151 patients with HCC were included, comprising 72 with NAFLD-related HCC and 79 with hepatitis B virus (HBV) cirrhosis–related HCC. On Sonazoid-CEUS, more than half of the NAFLD-related HCCs exhibited relatively early and mild washout within 60 seconds (54.2%, 39/72), whereas most HBV cirrhosis–related HCCs displayed washout between 60 and 120 seconds (46.8%, 37/79) or after 120 seconds (39.2%, 31/79) (P<0.001). In the patients with NAFLD-related HCC, multivariate analysis revealed that international normalized ratio (odds ratio [OR], 0.002; 95% confidence interval [CI], 0.000 to 0.899; P=0.046) and poor tumor differentiation (OR, 21.930; 95% CI, 1.960 to 245.319; P=0.012) were significantly associated with washout occurring within 60 seconds. Conclusion Characteristic Sonazoid-CEUS features are useful for diagnosing HCC in patients with NAFLD.