Objective This study aimed to investigate the effects of sleep interruption(SI)cycles on emotional behavior in ICR mice,and to establish a mouse model of comorbid anxiety and depression induced by SI.Methods Seventy-two male ICR mice(4～5 weeks old)were divided randomly into a blank group and a model group.Mice in the model group were subjected to SI stress modeling for 1,2,and 3 weeks,respectively.After modeling,emotional behaviors were evaluated using open-field,elevated plus maze,light-dark box,marble-burying,and forced-swimming tests.Serum corticosterone levels were detected by enzyme-linked immunosorbent assay.Results Mice in the model group buried significantly more marbles after 1 week of SI stress,compared with the blank group(P＜0.05).After 2 weeks of stress,mice in the model group also showed a significant decrease in the number of crossings in the light-dark box(P＜0.05)and a significant increase in the number of marbles buried(P＜0.01)compared with the control group.After 3 weeks of stress,mice in the model group showed a significant increase in the number of marbles buried(P＜0.05),a significant decrease in the number of crossings in the light-dark box(P＜0.05),and a significant increase in immobility time in the forced-swim test(P＜0.01).Conclusions ICR mice exhibited significant anxiety-related behaviors after 2 weeks of SI modeling and significant anxiety-and depressive-related behavioral changes after 3 weeks.Three weeks of SI stress can be used to establish a model of comorbid anxiety and depression.

Objective This study aimed to investigate the effects of sleep interruption(SI)cycles on emotional behavior in ICR mice,and to establish a mouse model of comorbid anxiety and depression induced by SI.Methods Seventy-two male ICR mice(4～5 weeks old)were divided randomly into a blank group and a model group.Mice in the model group were subjected to SI stress modeling for 1,2,and 3 weeks,respectively.After modeling,emotional behaviors were evaluated using open-field,elevated plus maze,light-dark box,marble-burying,and forced-swimming tests.Serum corticosterone levels were detected by enzyme-linked immunosorbent assay.Results Mice in the model group buried significantly more marbles after 1 week of SI stress,compared with the blank group(P＜0.05).After 2 weeks of stress,mice in the model group also showed a significant decrease in the number of crossings in the light-dark box(P＜0.05)and a significant increase in the number of marbles buried(P＜0.01)compared with the control group.After 3 weeks of stress,mice in the model group showed a significant increase in the number of marbles buried(P＜0.05),a significant decrease in the number of crossings in the light-dark box(P＜0.05),and a significant increase in immobility time in the forced-swim test(P＜0.01).Conclusions ICR mice exhibited significant anxiety-related behaviors after 2 weeks of SI modeling and significant anxiety-and depressive-related behavioral changes after 3 weeks.Three weeks of SI stress can be used to establish a model of comorbid anxiety and depression.

Male infertility is a significant cause of psychosocial and marital distress in approximately 50% of couples who are unable to conceive, with male factors being the underlying cause. Guijiajiao (Colla Carapacis et Plastri, CCP) is a Traditional Chinese Medicine commonly used to treat male infertility. The present study aimed to investigate the potential mechanisms underlying the preventive effects of CCP on male infertility. An infertile male rat model was established using cyclophosphamide (CTX), and CCP was administered for both treatment and prevention. Fecal microbiota transplantation (FMT) was also performed to explore the role of gut microbiota in the CCP-mediated prevention of male infertility in rats. Sperm motility and concentration were determined using a semi-automatic sperm classification analyzer. Subsequently, histopathological analysis using HE staining was performed to examine the changes in the small intestine and testis. Moreover, the serum levels of lipopolysaccharide (LPS) and testosterone were measured by ELISA. In addition, immunohistochemistry was conducted to detect CD3 expression in the small intestine, while RT-qPCR was employed to assess the expressions of interleukin-1 beta (IL-1β), cluster of differentiation 3 (CD3), Monocyte chemoattractant protein-1 (MCP-1), and C-X-C motif chemokine ligand 10 (CXCL-10) in the small intestine and epididymis. Finally, gut microbiota was analyzed by 16S rRNA sequencing. CCP improved sperm motility, number, and concentration in CTX-induced infertile male rats. CCP increased the serum testosterone level, inhibited the immune cell infiltration of the intestinal lamina propria, and promoted the aggregation of CD3⁺ T cells in CTX-induced male infertility rats. CCP also inhibited the expressions of MCP-1, CXCL-10, and IL-1β in the epididymis of male infertility rats. At the genus level, CTX led to a reduction in the abundance of Lactobacillus, Clostridia_UCG.014, and Romboutsia in the intestinal tract of rats. In contrast, CCP decreased the abundance of Ruminococcus and increased the abundance of Romboutsia in infertile male rats. Additionally, FMT experiments proved that the gut microbiota of CCP-treated rats facilitated testicular tissue recovery and spermatogenesis while also reducing the serum LPS level in infertile male rats. CCP improves the spermatogenic ability of infertile male rats by restoring gut microbiota diversity and inhibiting epididymal inflammation.
Humans ; Rats ; Male ; Animals ; Gastrointestinal Microbiome ; Lipopolysaccharides/pharmacology* ; RNA, Ribosomal, 16S ; Semen ; Sperm Motility ; Infertility, Male/prevention & control* ; Testosterone

The essence-qi-spirit theory is an important part of traditional Chinese medicine， whose steady state is the material and functional basis for the balance of yin and yang in the body， making the essence， qi and spirit integrated， and body and spirit harmonized. Based on this theory， it is proposed that essence and qi depletion， spirit dissipation and qi dispersion， disharmony between yin and yang is the main pathogenesis of sleep disorders. Therefore， the method of regulating and harmonzing yin and yang by essence gathering， qi nourishing and spirit storing can be used to treat sleep disorder. The biological clock system of the circadian rhythm of sleep is regulated by the molecular oscillation that is generated by the transcription of the biological clock gene， and is a clock gradually formed by orga-nisms constantly adapting to the laws of nature. As the material basis， power， and embodiment of sound and peaceful sleep， essence， qi and spirit can perceive and transmit natural signals， whose functions are similar to what is recognized by modern science that oscillation amplifies the rhythm signal， and synchronously regulates the expression signal of the biological clock gene， thereby forming a biological clock system with “input-oscillation-output” as the feedback cycle. It is believed that the regulation method of yin and yang by essence gathering， qi nourishing and spirit storing may comprehensively regulate the physiological activities through brain/ muscle aryl hydrocarbon receptor nuclear translocator-like protein 1 （BMAL1）/circadian locomotor output cycles kaput （CLOCK）-period protein （PER）/ cryptochrome （CRY） transcriptional feedback loop， thereby adapting to the natural environment changes， playing an active role in the treatment of sleep disorders， and provideing a new idea for traditional Chinese medicine to reshape the molecular regulation system of the endogenous biological clock to prevent and treat sleep disorders.

OBJECTIVE Microgravity exerts several negative effects on the learning and memory of astro-nauts during space flight.Rg1 and Rb1,the key steroidal components of ginseng,have shown potent neuroprotec-tive effects with a high safety profile.The object of the current study is to investigate the influence of Rg1 and Rb1 on simulated microgravity-induced memory and learning dysfunction in the hindlimb suspension(HLS)rat model.METHODS The HLS rats were orally administered Rg1(30 and 60 μmol·kg-1)or Rb1(30 and 60 μmol·kg-1)for four weeks.The Morris water maze test(MWM)and reward operating conditioning reflex test(ROCR)were conducted to evaluate spatial and associative learning and memory.After the behavior tests,the serum and the prefrontal cortex(PFC)were dissected to measure the mechanism.RESULTS Rg1 and Rb1 treatment amelio-rated the cognitive deficits of HLS-exposure rats in MWM and ROCR,reduced reactive oxygen species generation and increased antioxidant enzyme activity.Rg1 and Rb1 also assisted in the recovery of mitochondrial complex Ⅰ(NADH dehydrogenase)activities and Mfn2,and decrea-sed Drp-1 expression.Furthermore,Rg1 and Rb1 reduced the ratio of Bax/Bcl-2 and the expression of cleaved-cas-pase 3,cytochrome c,increased the levels of SYN,PSD95 and activated BDNF-TrkB/PI3K-Akt pathway in the PFC.CONCLUSION Rg1 and Rb1 treatment attenuated cog-nitive deficits induced by HLS,mitigated mitochondrial dysfunction,attenuated oxidative stress,inhibited apopto-sis,and increased the synaptic plasticity,which was partly mediated by the modulation of the BDNF-TrkB/PI3K-Akt signaling.

The most common manifestations of inflammatory bowel disease （IBD） are Crohn's disease and ulcerative colitis， and the global incidence of IBD is on the rise. Traditional Chinese medicine （TCM） is advantageous in the treatment of IBD. IBD， with TCM names based on clinical symptoms， mostly belongs to recurrent dysentery， long dysentery， diarrhea， dysentery， bowel， and other categories. In TCM pathogenesis of IBD， spleen deficiency and exuberant dampness predominate in the whole course of the disease. Since the lung is associated with the large intestine and the lung Qi and spleen Qi are interconnected， the lung Qi and spleen Qi are deficient and the dampness and heat accumulate internally， which caused collateral obstruction by stagnant blood and the development of IBD. From the perspective of "associating lung with large intestine"，it is believed that the main mechanism of IBD is the Qi imbalance and abnormal metabolism of fluids in the lung and the intestine，and the nutrient-Yin injury of the lung and the intestine. According to the chronic， recurrent， and diffuse pathogenesis characteristics and main clinical manifestations of IBD， IBD is closely related to the lung and the intestine. In terms of therapeutic principles， IBD can be treated by tonifying the spleen and replenishing the lung， which highlights the treatment of the intestine from the lung. To be specific， in time of tonifying the spleen and removing dampness， the intestine is regulated by tonifying the spleen and replenishing the lung. Shenling Baizhusan， a commonly used classical prescription for IBD， is mainly potent in replenishing Qi， invigorating the spleen， draining dampness， checking diarrhea， and especially "reinforcing earth to generate metal". It can enhance the function of the lung through "reinforcing earth to generate metal"， which in turn regulates the intestine and promote the improvement of IBD. The present study clarified the mechanism of Shenling Baizhusan in regulating the intestine by tonifying the spleen and replenishing the lung. On the basis of modern research， its therapeutic effect on IBD was achieved through multiple links， such as regulation of the level of inflammatory factors， immunoregulation， barrier function improvement via mucosal repair， and intestinal flora. The findings of this study are expected to provide new ideas for the regulation of the lung-spleen-large intestine axis in the syndrome differentiation and treatment of IBD and subsequent experimental research.

ObjectiveTo explore the effect of Guilu Erxianjiao on improving reproductive injury in diabetic rats and its possible mechanism. MethodFifty-three SD male rats were randomly divided into 5 groups， with 1 group as the normal group and the other 4 groups as the modeling groups. Rats in the modeling groups were fed with a high-fat diet combined with 30 mg·kg^-1 streptozotocin （STZ） intraperitoneal injection to induce diabetes， with the random blood glucose >16.7 mmol·L^-1 for 3 consecutive times as the criteria for inclusion in the model of diabetic reproductive injury. The rats with diabetic reproductive injury were then randomly divided into a model group， a Guilu Erxianjiao group （2 g·kg^-1）， a Vitamin E group （0.03 g·kg^-1）， and a Wuzi Yanzong pill group （0.6 g·kg^-1） according to the blood glucose level. The rats were given the corresponding drug dose intragastric administration， once a day for 4 weeks， and their body weight and blood glucose were measured weekly. After 4 weeks， samples were collected for index determination. Morphological changes in testis and epididymis were observed by hematoxylin-eosin （HE） staining， and apoptosis of testis cells was observed by in situ end labeling （TUNEL） staining. Sperm concentration and motility were detected by the semen analyzer. Serum levels of follicle-stimulating hormone （FSH）， luteinizing hormone （LH）， and testosterone （T） were determined by enzyme-linked immunosorbent assay （ELISA）. The content of superoxide dismutase （SOD）， propylene glycol （MDA）， reactive oxygen species （ROS）， and glutathione peroxidase （GSH-Px） in the testicular tissue was determined by ELISA. The expressions of nuclear respiratory factor-2 （Nrf2）， heme oxygenase 1 （HO-1）， B-cell lymphoma-2 （Bcl-2）， and Bcl-2 associated X protein （Bax） in the testicular tissue were detected by Western blot. ResultAs compared with the normal group， testicular tissue atrophy， decreased spermatogenic tubules， epididymal wall hyperplasia， and lumen stenosis were observed in the model group. Sperm concentration and motility decreased （P<0.01）， and serum levels of T， FSH， and LH decreased （P<0.01） in the model group. The content of ROS and MDA in the testis increased （P<0.01）， while that of SOD and GSH-Px decreased （P<0.01） in the model group. The expression of Bax increased （P<0.01）， and the expressions of Nrf2， HO-1， and Bcl-2 decreased （P<0.01） in the model group. As compared with the model group， the pathological changes in the testis and epididymis in the Guilu Erxianjiao group were improved to some extent. Sperm concentration and motility increased （P<0.05， P<0.01）. In the Guilu Erxianjiao group， serum levels of T and LH increased （P<0.05， P<0.01）， while FSH levels showed no significant difference. The content of ROS and MDA in the testis decreased （P<0.01）， while that of SOD and GSH-Px increased （P<0.01） in the Guilu Erxianjiao group. The expression of Bax decreased （P<0.01）， and the expressions of Nrf2， HO-1， and Bcl-2 increased （P<0.05， P<0.01） in the Guilu Erxianjiao group. ConclusionGuilu Erxianjiao improves the reproductive injury and sperm quality of diabetic rats to a certain extent， and the mechanism may be related to the improvement of oxidative stress and anti-apoptosis.

Objective:To analyze the imaging features of hepatic metastases in gastrointestinal stromal tumors (GIST) on 18F-fluorodexyglucose (FDG) PET/CT in order to improve the accuracy of diagnosis. Methods:Clinical and imaging data of 33 patients (18 males, 15 females, age 34-70 years) with hepatic metastases in GIST who underwent PET/CT examination between May 2013 and July 2019 in Fujian Cancer Hospital were analyzed retrospectively. All patients underwent 18F-FDG PET/CT early imaging, and nine of them underwent delayed imaging. Visual analysis was performed on the PET/CT images by comparing FDG uptake of hepatic lesions and liver background, and all lesions were classified as significant hypermetabolism, slightly higher metabolism and equal or lower metabolism. Maximum standardized uptake value (SUV max) of primary GIST lesions and hepatic metastases were calculated and compared, and the relationship between them was analyzed. Wilcoxon rank sum test and Spearman rank correlation analysis were used to analyze the data. Results:Among 33 GIST patients, 9 patients had solitary hepatic metastasis, and 24 patients had multiple hepatic metastases (104 lesions). The diameter of metastases was 0.8-14.6(2.2(1.5, 3.9)) cm, and SUV max was 1.4-21.5(3.6(2.4, 5.7)). Of the 104 hepatic metastases, 94.2%(98/104) lesions had clear boundaries, 65.4%(68/104) lesions had uniform density (2 lesions with cystic density), 34.6%(36/104) lesions had uneven density in which hemorrhage, cystic change or necrosis could be found. On visual analysis of PET images, 38.5%(40/104) lesions were with significant hypermetabolism, 26.0%(27/104) were with slightly higher metabolism and 35.6%(37/104) were with equal or lower metabolism. In 24 patients with multiple hepatic metastases, 79.2%(19/24) showed different metabolic levels synchronously. Among 67 hypermetabolic metastases, 34.3%(23/67) were with homogeneous metabolism, of which 13 lesions with diameter<2.0 cm; 65.7%(44/67) were with heterogeneous metabolism, of which 36 lesions with diameter≥2.0 cm. There was a moderate correlation of SUV max between GIST primary tumors and hepatic metastases ( n=15; 9.2(6.8, 14.5) vs 3.8(2.1, 6.0), rs=0.556, P<0.01). The difference of SUV max between GIST primary tumors and hepatic metastases was statistically significant ( z=-5.098, P<0.01). In delayed imaging, 13/15 hepatic metastases with equal or lower metabolism changed to slightly higher metabolism. Conclusions:Hepatic metastases in GIST on 18F-FDG PET/CT imaging usually have clear boundary, and often associate with cystic degeneration, hemorrhage or necrosis. The metabolic patterns of hepatic metastases in GIST are varied. Delayed PET/CT imaging is helpful for the diagnosis of hypometabolic hepatic metastases in GIST.

Objective To compare the diagnostic values of 18F-FDG and 18F-FLT PET/CT in patients with suspicious recurrence of glioma after multimodal treatment.Methods A total of 20 patients (13 males,7 females;age range:12-73 years) with glioma who underwent 18F-FDG and 18F-FLT PET/CT due to abnormal enhancement on MRI from January 2012 to June 2015 were enrolled in this retrospective study.According to the pathological or follow-up results,patients were divided into therapy-related benign changes (TRBC) group and recurrent glioma group,the later was subdivided into initial low-grade glioma (LGG) group and initial high-grade glioma(HGG) group.T/NT ratios of 18F-FDG and 18F-FLT between HGG (LGG) group and TRBC group were compared using one-way analysis of variance and the least significant difference t test.ROC curve analysis was conducted to calculate the differential diagnostic efficiency of 18F-FDG and 18F-FLT between TRBC and recurrent glioma.Results A total of 14 patients were proved as recurrent glioma and 6 patients as TRBC.The mean 18F-FDG T/NT ratios of HGG group,LGG group and TRBC group were 2.31±0.86,1.32±0.86 and 1.32±0.64,respectively.The 18F-FDG T/NT ratio of the HGG group was significantly higher than that of the TRBC group(F=3.671,t=-2.471,P＜0.05).The mean 18F-FLT T/NT ratios of HGG group,LGG group and TRBC group were 8.94±3.14,7.18±3.29 and 1.92±1.20,respectively (F=13.301,t values:-5.150 and-2.360,both P＜0.05).The optimal T/NT cutoff values for 18 F-FDG and 18F-FLT PET/CT were 1.62 and 4.58,respectively.The sensitivity,specificity and accuracy of detecting recurrent glioma with optimal T/NT cutoff value were 11/14,5/6 and 16/20 for 18F-FDG PET/CT,and those for 18F-FLT PET/CT were 13/14,6/6 and 19/20,respectively.No significant difference was observed between the diagnostic efficiencies of the two imaging modalities (x2 values:1.167,1.091 and 2.057;all P＞0.05).Conclusion There were no statistical significances between 18F-FDG and 18F-FLT PET/CT on the differential diagnosis of glioma recurrence.

Objective: To evaluate the value of ¹⁸F-FDG PET-CT in predicting the malignant potential of Gastrointestinal Stromal Tumors (GIST). Methods: The clinical and pathological features of 31 patients with GIST confirmed by surgery or biopsy were retrospectively analyzed. The malignant potential of GIST before treatment was assessed by ¹⁸F-FDG PET-CT. The GIST risk classification was graded according to the Standard revised by the National Institutes of Health (NIH) in 2008. The relationship between the maximal standard uptake value (SUVmax) and GIST risk classification, tumor diameter, Ki-67 index, and mitotic count were analyzed respectively. The cut-off level of SUVmax for the diagnosis of malignant GIST was calculated from the Receiver Operating Characteristic (ROC) curve. Results: Among the 31 cases of GIST patients, 14 cases were gastric primary (stomach group) and 17 cases were nongastric primary (outside stomach group). The SUVmax, tumor diameter, Ki-67 index and mitotic count of the 31 patients were 8.21±4.68, (7.82±5.12)cm, (10.03±11.07)% and (12.29±10.55)/50 HPF, respectively. SUVmax was significantly correlated with GIST risk classification (r=0.727, P<0.01), but not with tumor diameter, Ki-67 index and mitotic count (r=0.348, r=0.284, r=0.290, P=0.055, P=0.121, P=0.114). The SUVmax, tumor diameter, Ki-67 index and mitotic count in the stomach group were 4.36±2.36, (6.08±4.31)cm, (3.43±3.03)% and (5.71±2.20)/50 HPF, respectively. SUVmax was significantly correlated with tumor diameter, GIST risk classification and Ki-67 index (r=0.682, r=0.868, r=0.732, P<0.01) but not with mitotic count (r=0.510, P=0.063). The SUVmax of the GIST in the gastric group and the outside gastric group were 4.36±2.36 and 10.68±5.50, respectively. The difference was statistically significant (P=0.001). The SUVmax in the malignant group of GIST (middle or high risk grade) was 8.90±4.89, which was significantly higher than 2.22±0.86 in the benign group (low or very low risk grade). The difference was statistically significant between the two group (P<0.01). ROC curve analysis showed that a SUVmax cut-off of 3.75 was the most sensitive for predicting malignant GIST. When the area under the curve of 0.969, the sensitivity was 84.6% and the specificity was 100%. Conclusions The SUVmax was strongly correlated with the GIST risk category and also with the tumor diameter and Ki-67 index in the gastric primary GIST, so it can be used as an effective indicator in predicting malignant potential of GIST before treatment.