Peroxisomes (microbodies) were studied with CATase, G-6-Pase, TPPase, CMPase cytochemistry and stereology by electron microscopy in 15 cases of normal human liver specimens. In human hepatocytes peroxisomes are ellipsoid or round intracellular bodies. They contain homogeneous fine granular matrix, and surrounded by unit membrane, and distribute randomly in the cytoplasm. By stereological analysis, the peroxisomes accounted for 1.33?0.38% of the cytoplasmic volume, their numerical density in spase was 9.76?1.63/100 ?m~3 cytoplasm, and the ratio of mitochondria to peroxisomes in quantity was 6.22?1.17. The result of G-6-Pase cytochemistry showed no luminal connection between endoplasmic reticulum and peroxisomes. The membrane similar to smooth endoplasmic reticulum seem to be connected with peroxisomes which was G-6-Pase negative, hence such membrane may be peroxisomal membrane system rather than smooth endoplasmic reticulum. The TPPase cytochemistry showed that peroxisomes did not connect with the Golgi complex structurally, and the Golgi complex may not be involved in the biogenesis of peroxisomes. Some figures resembled fission and budding of peroxisomes were viewed, and this finding suggested that new peroxisomes formed probably by division of preexisting peroxisomes.

OBJECTIVE: To investigate the effect of inner-heating acupuncture on apoptosis of chondrocytes and expression of Caspase-3 and Caspase-9 in rats with knee osteoarthritis (KOA). METHODS: A total of 32 rats were divided into a normal group, a model group, a control treatment group and a treatment group by random number grouping method, 8 rats in each one. The rats in the normal group received no intervention. The rats in the remaining three groups adopted modified Videman method to develop KOA model, the ankle joint of left posterior leg was fully extended and fixed with a resin bandage for 6 weeks. After successful modeling, the rats in the model group received no intervention. The rats in the control treatment group were treated with medium-frequency pulse electrotherapy. The rats in the treatment group were treated with inner- heating acupuncture, 30 min each treatment, once a day, five days per week, and totally 3-week treatment was given. After 3 weeks, the damaged cartilage tissue was collected, and HE staining was used to observe the pathological changes of the cartilage tissue of the knee joint. ELISA was used to detect the content of cytochrome-C in the tissue homogenate supernatant. The chondrocytes in damaged cartilage tissue were isolated, flow cytometer was used to detect the changes of apoptosis and mitochondrial membrane potential. The mRNA and protein expression of Caspase-3 and Caspase-9 in chondrocytes were detected by real-time quantitative PCR (qRT-PCR) and Western blot (WB), respectively. RESULTS: Compared with the normal group, the damage of cartilage tissue in the model group was significant, and the expression level of Cyt-C in the homogenate supernatant of damaged cartilage tissue was increased (<0.01); the chondrocyte apoptosis was increased significantly (<0.01); the chondrocyte mitochondrial membrane potential was decreased significantly (<0.01); the mRNA and protein expression of Caspase-3 and Caspase-9 was increased significantly (all <0.01). Compared with the model group, the cartilage injury in the control treatment group and the treatment group was significantly relieved; the expression level of Cyt-C in the supernatant of damaged cartilage tissue homogenate was decreased (both <0.01); the chondrocyte apoptosis was significantly reduced (both <0.01); the chondrocyte mitochondrial membrane potential was increased significantly (both <0.01). Moreover, the mRNA and protein expression of Caspase-3 and Caspase-9 was significantly reduced (all <0.01). Compared with the control treatment group, the treatment group was more effective in the treatment of KOA. CONCLUSION The inner-heating acupuncture could significantly improve the pathological changes of KOA rats, inhibit the apoptosis of chondrocytes, which may be closely related to the suppression of Caspase-3 and Caspase-9 expression.
Animals ; Apoptosis ; Cartilage, Articular ; Caspase 3 ; Caspase 9 ; Chondrocytes ; Heating ; Osteoarthritis, Knee ; Rats

Objective To investigate the effect of interferon alpha ( IFN-α) on apoptosis of vascular smooth muscle cells ( VSMCs) in rats and the related mechanism. Methods The cells were divided into three group:group A, group B and group C.Group A was transfected with nonspecific siRNA, group B was intervened with IFN-α and transfected with nonspecific siRNA, and group C was intervened with IFN-α and transfected with IFI204 siRNA. All the cells were cultured for 48 h. The expression of P204 mRNA was determined by semiquantitative reverse transcription polymerase chain reaction (RT-PCR).P204, RAS protein levels, and phosphorylation levels of RAF and ERK were analyzed by Western blotting. The cell apoptosis was analyzed by flow cytometry with Annexin-V FITC/PI method. Results As compared with group A, the expression of P204 mRNA and protein in group B was up-regulated (P<0.05), the cell apoptosis was increased (P<0.05), in the process of the above, the expression of RAS protein was decreased ( P<0.05) and the phosphorylation levels of RAF and ERK were dropped (P<0.05).In group C, the expression levels of P204 mRNA and protein were down-regulated (P<0.05), and cell apoptosis was decreased ( P<0.05) , the expression of RAS protein and the phosphorylation levels of RAF and ERK were increased ( P<0.05) . Conclusion P204 and RAS signal pathway participates in IFN-α regulation of apoptosis of VSMCs in rats.

OBJECTIVE: To investigate the effect of γ-secretase inhibitor DAPT in inflammation-induced brain white matter injury in neonatal mice. METHODS: Sixty C57BL/10J neonatal mice are randomly divided into control group, control+DAPT (10 mg•kg-1) group, inflammation (LPS) group, LPS+DAPT group (inflammation exposure after 10 mg•kg-1 DAPT treatment). All neonatal mice were killed and brain was removed to the following observation and detection:at P5, the mRNA expression variation of IL-1β, IL-8,TNF-α,Hes1 and NICD by Real-time PCR methods. Oligodendrocytes were identified by immunofluorescence staining. Myelin basic protein (MBP) protein expression was detected by Western blot assay. RESULTS: LPS group showed brain injury characterized by inhibition of brain development. There were significant differences in mRNA expression of IL-1β, IL-8, TNF-α, Hes1 and NICD between LPS+DAPT group and LPS group (P<0.05), and the mRNA expression of IL-1β, IL-8,TNF-α,Hes1 and NICD in inflammation-treated were significantly increased than control group (P<0.05). The results showed more expression of MBP in LPS+DAPT group compared with LPS group (P<0.05). Compared with the blank control group, which was obviously decreased after 48 h of inflammation (P<0.05).CONCLUSION: Inflammation leads to abnormal of notch signal expression in neonatal mice, and which is shows inflammation involved in brain damage.Its mechanism is probably associated with the maturation of oligodendrocytes.

BackgroundPrevious studies have found that patients with generalized anxiety disorder （GAD） have impaired performance in executive function， and group cognitive behavioral therapy （CBT） has been shown to be effective in alleviating negative affect in patients with GAD， while its efficacy on executive function remains unclear. ObjectiveTo explore the efficacy of group CBT on anxiety symptom and executive function in GAD patients， so as to provide references for the rehabilitation program for GAD. MethodsA total of 80 consecutive patients with GAD who were hospitalized in Sleep and Psychosomatic Medical Center of Shiyan Taihe Hospital from March 2021 to August 2022 and met the Diagnostic and Statistical Manual of Mental Disorders， fifth edition （DSM-5） diagnostic criteria for GAD were enrolled， and they were assigned into study group （n=40） and control group （n=40） using random number table methods. All patients were subjected to routine medication treatment and regular health education， based on this， study group received group CBT once a week （6 weeks， 60 to 90 minutes per session）. At the enrollment and after 6 weeks of treatment， patients were assessed using Hamilton Anxiety Scale （HAMA） and Frontal Assessment Battery （FAB）. ResultsANOVA with repeated measures on HAMA score revealed a significant time effect （F=1 870.320， P<0.01）， no significant group effect and no significant time×group interaction effect （F=1.254， 0.293， P>0.05）. Significant time effect， group effect and time×group interaction effect were reported on FAB scores （F=311.190， 4.399， 7.021， P<0.05 or 0.01）. Further analysis indicated that FAB scores of both groups after treatment were higher than those at baseline （t=200.569， 115.401， P<0.01）.And the FAB score of study group was higher than that of control group after treatment （t=-3.211， P<0.01）. ConclusionGroup CBT combined with medication treatment for GAD may alleviate the anxiety symptoms and improve executive function in GAD patients. ［Funded by Shiyan Science and Technology Bureau Pilot Scientific Research Project （number， 21Y21）］

OBJECTIVETo study effects of electroacupuncture at Jiaji (EX-B2) on protracted withdrawl syndrome of the patient of heroin dependence.

METHODSOne hundred and twenty cases of heroin dependence were randomly divided into 4 groups: acupuncture group I (Jianji and Shenshu acupoints), acupuncture group II (acupoints at limbs), simulation group and control group. Protracted withdrawl syndrome scale, Hamilton anxiety scale (HAMA) and self-rating depression scale (SDS) were used to observe changes of the scores before and after treatment of 4, 8, 10 weeks.

RESULTSIn the treatment of 4, 8, 10 weeks, the cumulative scores for protracted withdrawl syndrome, HAMA and SDS in the acupuncture group I and II decreased significantly as compared with those in the control group (P < 0.01).

CONCLUSIONElectroacupuncture can significantly improve protracted withdrawl syndrome, alleviate anxiety and depression, and electroacupuncture at Jiaji (EX-B2) being better than at the acupoints of the limbs.

Acupuncture Points ; Acupuncture Therapy ; Electroacupuncture ; Heroin Dependence ; Humans ; Substance Withdrawal Syndrome

OBJECTIVETo investigate the correlation between the ratio of serum vascular endothelial growth factor (VEGF)/endostatin (ES) and childhood acute leukemia(AL).

METHODSSerum levels of VEGF and ES were measured using ELISA in 35 children with acute AL before and after chemotherapy. The ratio of VEGF/ES was calculated. Thirty healthy children served as the control group.

RESULTSThe serum levels of VEGF (196 ± 66 pg/mL vs 29 ± 10 pg/mL) and ES (35 ± 7 ng/mL vs 19 ± 4 ng/mL) in the AL group before chemotherapy were significantly higher than those in the control group (P<0.01). The ratio of VEGF/ES in the AL group before chemotherapy was significantly higher than that in the control group (6.7 ± 3.0 vs 1.6 ± 0.7; P<0.01). In 20 children with AL who achieved complete remission, the serum levels of VEGF and ES and the VEGF/ES ratio were reduced after chemotherapy (83 ± 35 pg/mL, 27 ± 5 ng/mL, 3.1 ± 1.3, respectively; P<0.01), although the serum levels of VEGF and ES were still higher than those in the control group (P<0.01). The VEGF/ES ratio in CR patients was not significantly different from that in the control group. The serum levels of VEGF (r=0.301, P=0.045) and the VEGF/ES ratio (r=0.411, P=0.015) were positively correlated with the count of blast cells in juvenile bone marrow in 35 children with AL before chemotherapy.

CONCLUSIONSSerum VEGF and ES levels are associated with the development of childhood AL. The VEGF/ES ratio can be used to evaluate the disease progression in children with AL.

Acute Disease ; Adolescent ; Child ; Child, Preschool ; Endostatins ; blood ; Female ; Humans ; Leukemia ; blood ; drug therapy ; Male ; Vascular Endothelial Growth Factor A ; blood

BACKGROUND: Sweat secreted by eccrine sweat glands is transported to the skin surface through the lumen. The eccrine sweat gland develops from the initial solid bud to the final gland structure with a lumen, but how the lumen is formed and the mechanism of lumen formation have not yet been fully elucidated. This study aimed to investigate the mechanism of lumen formation of eccrine gland organoids (EGOs). METHODS: Human eccrine sweat glands were isolated from the skin for tissue culture, and the primary cultured cells were collected and cultured in Matrigel for 14 days in vitro. EGOs at different development days were collected for hematoxylin and eosin (H&E) staining to observe morphological changes and for immunofluorescence staining of proliferation marker Ki67, cellular motility marker filamentous actin (F-actin), and autophagy marker LC3B. Western blotting was used to detect the expression of Ki67, F-actin, and LC3B. Moreover, apoptosis was detected using a terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) apoptosis assay kit, and the expression of poly (ADP-ribose) polymerase and Caspase-3 was detected by Western blot. In addition, 3-methyladenine (3MA) was used as an autophagy inhibitor to detect whether the formation of sweat glands can be effectively inhibited. RESULTS: The results showed that a single gland cell proliferated rapidly and formed EGOs on day 4. The earliest lumen formation was observed on day 6. From day 8 to day 14, the rate of lumen formation in EGOs increased significantly. The immunofluorescence and Western blot analyses showed that the expression of Ki67 gradually decreased with the increase in days, while the F-actin expression level did not change. Notably, the expression of autophagy marker LC3B was detected in the interior cells of EGOs as the apoptosis signal of EGOs was negative. Compared with the control group, the autophagy inhibitor 3MA can effectively limit the formation rate of the lumen and reduce the inner diameter of EGOs. CONCLUSION Using our model of eccrine gland 3D-reconstruction in Matrigel, we determined that autophagy rather than apoptosis plays a role in the lumen formation of EGOs.
Apoptosis ; Autophagy ; Eccrine Glands ; Epithelial Cells ; Humans ; Organoids

Humans ; Nasal Cavity ; Melanoma ; Skin Neoplasms ; Paranasal Sinuses

Acute lung injury (ALI) is a prevalent and severe clinical condition characterized by inflammatory damage to the lung endothelial and epithelial barriers, resulting in high incidence and mortality rates. Currently, there is a lack of safe and effective drugs for the treatment of ALI. In a previous clinical study, we observed that Jinyinqingre oral liquid (JYQR), a Traditional Chinese Medicine formulation prepared by the Taihe Hospital, Affiliated Hospital of Hubei University of Medicine, exhibited notable efficacy in treating inflammation-related hepatitis and cholecystitis in clinical settings. However, the potential role of JYQR in ALI/acute respiratory distress syndrome (ARDS) and its anti-inflammatory mechanism remains unexplored. Thus, the present study aimed to investigate the therapeutic effects and underlying molecular mechanisms of JYQR in ALI using a mouse model of lipopolysaccharide (LPS)-induced ALI and an in vitro RAW264.7 cell model. JYQR yielded substantial improvements in LPS-induced histological alterations in lung tissues. Additionally, JYQR administration led to a noteworthy reduction in total protein levels within the BALF, a decrease in MPAP, and attenuation of pleural thickness. These findings collectively highlight the remarkable efficacy of JYQR in mitigating the deleterious effects of LPS-induced ALI. Mechanistic investigations revealed that JYQR pretreatment significantly inhibited NF-κB activation and downregulated the expressions of the downstream proteins, namely NLRP3 and GSDMD, as well as proinflammatory cytokine levels in mice and RAW2647 cells. Consequently, JYQR alleviated LPS-induced ALI by inhibiting the NF-κB/NLRP3/GSDMD pathway. JYQR exerts a protective effect against LPS-induced ALI in mice, and its mechanism of action involves the downregulation of the NF-κB/NLRP3/GSDMD inflammatory pathway.
Humans ; NF-kappa B/metabolism* ; Lipopolysaccharides/metabolism* ; NLR Family, Pyrin Domain-Containing 3 Protein/metabolism* ; Acute Lung Injury/metabolism* ; Lung ; Phosphate-Binding Proteins/therapeutic use* ; Pore Forming Cytotoxic Proteins/therapeutic use*