Objective To establish chronic sleep deprivation mouse model,evaluate the learning and memory ability of mice and observe autophagy and apoptosis levels in mouse brain.Methods C57BL/6 mice (n =20) were randomly separated into sleep deprivation group and control group.After 2-month sleep deprivation by using an adapted multiple platform method,the behavioral performance of mice was measured by IntelliCage system.The expression of microtubule associated protein 1 light chain 3-Ⅱ (LC3-Ⅱ) and Beclin-1 was detected by Western blotting.Confocol microscopy was used to observe autophagosome.In addition,terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) staining was performed to detect neuronal apoptosis level in mouse brain.Results The results of behavioral test showed that the incorrect visit ratio was much higher in sleep deprivation group than that in control group.Moreover,the expression of LC3-Ⅱ (sleep deprivation group 1.681 ± 0.186,control group 1.125 ±0.048,t =2.892,P =0.027 6) and Beclin-1 (sleep deprivation group 1.144 ±0.048,control group 1.006 ± 0.017,t =2.721,P =0.018 6) in mouse hippocampus and cortex was significantly elevated in sleep deprivation group than those in control group.Accordingly,the confocal microscopy observation also revealed an increased nuclear LC3-positive puncta in hippocampus and cortex of sleep deprived mice (hippocampus in sleep deprivation group 1.665 ± 0.153,in control group 0.819 ± 0.072,t =5.024,P =0.002 4;cortex in sleep deprivation group 1.925 ± 0.175,in control group 1.195 ± 0.111,t =3.521,P =0.012 5).In addition,TUNEL staining showed a much higher percentage of TUNEL-positive nuclei in these brain regions (hippocampus in sleep deprivation group 47.24 ± 4.15,in control group 19.26 ± 3.72,t =5.025,P =0.007 4;cortex in sleep deprivation group 42.25 ± 1.25,in control group 27.50 ± 3.23,t =4.262,P =0.005 3).Conclusions Chronic sleep deprivation can impair the learning and memory,increase the expression of LC3-Ⅱ and Beclin-1,elevate the formation of autophagosome,and promote apoptosis in mouse brain.These findings suggest that autophagy and apoptosis might be involved in the cognitive impairment induced by chronic sleep deprivation.

Objective To investigate the roles of nitric oxide and caspase 3 in PC12 cell apoptosis induced by rotenone. Methods The acute injury effect of rotenone on PC12 cells and viability of the PC12 cells were detected by MTT assay. Nitrite (NO 2 -) was quantified by Greiss reaction. The activity of caspase 3 and the cell apoptosis were detected by fluoremetry and agarose gel electrophoresis, respectively. Results The MTT colorimetry indicated that PC12 cell viability reached a peak at 7 d after passage. L NAME at the concentration of 100 ?mol/L showed the strongest protective effect for PC12 cell in the four different doses. Rotenone also induced the formation of DNA ladder in PC12 cells. Additionally, the nitric oxide synthase inhibitor L NAME inhibited the activity of caspase 3 protease. L NAME also inhibited the production of nitric oxide induced by rotenone. Conclusion Nitric oxide and caspase 3 are involved in the process of PC12 cell apoptosis induced by rotenone.

Objective To investigate lipopolysaccharide(LPS)-induced injury of dopaminergic neurons in the substantia nigra and effect of LPS on animals behavior in vitro and in vivo. Methods Pheochromocytoma(PC12)cell viability was estimated by MTT assay after cells had been differently incubated in medium containing LPS, 6-OHDA and culture supernatant of LPS-treated microglia. LPS was stereotactically injected into left substantia nigra of rats. 14,21 and 28 days after injection, the circling behavior was observed by intraperitoneal injection of apomorphine. Midbrain dopaminergic neurons were identified by using tyrosine hydroxylase (TH) immunohistochemistry. The contents of dopamine and its metabolites in the striatum and substantia nigra were measured by high performance liquid chromatography(HPLC). Results LPS alone had no influence on cell viability. However, the 26% and 30% reduction of cell viability was found when cells were exposed to culture supernatant of LPS-treated microglia and 6-OHDA. The circling behavior ipsilateral to LPS-treated side was induced in some rats 21 and 28 days following injection, but not 14 days. Compared with the PBS-treated rats, the marked (35%~60%) loss of TH-positive cells in left substantia nigra was identified 21 and 28 days after LPS treatment, but not 14 days. The 30%~70% reduction of dopamine and 3,4-dihydroxyphenylacetic acid (DOPAC) in left striatum and substantia nigra was found at 21 and 28 days following injection, whereas no significant change was observed at 14 days. Conclusions LPS could indirectly injury the dopaminergic neurons and induce behavioral asymmetry of rats.

AIM: To explore the mutual effect of co-culture of wild-type astrocytes (ASC) and motor neurons VSC4.1 (VSC) on the respective ability to produce reactive oxygen species (ROS). METHODS: The inhibition rates of cell growth in ASC and VSC in co-culture or independent culture was detected after exposed to excitatory stimulus by MTT method. Real-time observation of ROS production by ASC and VSC labeled with Hoechst 33342 was detected by confocal microscopy under the conditions of co-culture or independent culture. RESULTS: Higher concentration of glutamate induced a higher inhibition rate in mixed cell growth than that in ASC alone, while lower concentration of glutamate induced a higher inhibition rate in mixed cell growth than that in VSC only. Real-time observation by confocal microscopy showed that ROS production by VSC under the condition of co-culture, which showed a notable increase at 15 min, was significant less than that in independent culture, which peaked at 5 min and was gradually decreased. ROS production by ASC in co-culture began to increase significantly at 10 min. CONCLUSION: Compared to independent culture, ASC reduces the resting ROS production by co-cultured with VSC, while ASC prolongs the duration of ROS production by VSC after exposed to excitatory stimulus.

Objective:To investigate the effect of modified transanal approach in the repair of vesicorectal fistula after radical prostatectomy.Methods:From September 2011 to December 2019, 32 cases of vesicorectal fistula after radical prostatectomy were retrospectively analyzed. All patients underwent cystostomy before repair operation. The average diameter of the fistulas was 19 (3-40) mm. There was only one fistula in 24 cases and 8 cases with more than 2 fistulas. The operation was performed in the jack knife position, and the fistula was prepared by resection of the fistula through the anus with bipolar resectoscope. Then bladder wall and rectum wall were separated by the loop and sutured respectively. After operation, the patients were treated with antispasmodic and anti-infective treatment, and the catheter was retained. Cystography and cystoscopy were reexamined 3 months after operation. Catheter was removed in the successful cases, and the failure was repaired again.Results:All operations were completed successfully. The mean operation time was 67(55-125) min, and the median follow-up was 22 (6-30) months. Thirty-one cases (96.8%) were successfully repaired, of which 25 cases were successfully repaired at the first operation, and 6 cases were successfully repaired again (all by transanal route). One case failed to be repaired. He had received external pelvic radiotherapy before operation. After the failure of repair, cystoscopy showed large fistula and stiff surrounding tissue. Then bilateral ureteral skin stoma and cystectomy were performed.Conclusions:Modified transanal approach in the repair of vesicorectal fistula after radical prostatectomy is an effective method. This kind of operation has less trauma, fewer complications and can be operated repeatedly. It is suitable for patients with low position, small fistula and without radiotherapy.

Objective To investigate the clinical efficacy of levothyroxine replacement therapy in children with hypothyroidism.Methods 45 children with hypothyroidism admitted from Apr.2015 to Dec.2016 were selected as observation group,and 45 healthy children in the same period were selected as the control group.Children in the control group received regular diet and were paid attention to nutritional balance.Children in the observation group were given levothyroxine sodium on the basis of dietary guidance.The two groups were treated continuously for 6 months.The levels of thyroid function,insulin-like growth factor,indicators for growth and development and inflammatory factors were measured before and after treatment in both groups.Results After treatment for 3 months and 6 months,the levels of FT3,FT4,IGF-1,IGFBP-3 and IGF-1/IGFBP-3 in the observation group increased significantly compared with those before treatment,and the levels of TSH in the observation group decreased significantly(P＜0.05).The levels of FT3,lFT4,TSH,IGF-1,IGFBP-3 and IGF-1/IGFBP-3 in the observation group after treatment for 6 months were (7.17±0.06)pmol/L,(2.74±0.07)mU/L,(207.67±9.06)μg/L,(0.46±0.02) mg/L and(34.59±1.97).The difference was not statistically significant (P＞0.05).After 3 months and 6 months of treatment,the height,body weight,osteocalcin and 25-(OH)2D3 levels in the observation group were significantly higher than those in the control group (P＜0.05),and there was no significant difference between the two groups after 6 months (P＞0.05).The difference was statistically significant (P＜0.05).Conclusion Left thyroid hormone replacement therapy can significantly improve the thyroid function of children with hypothyroidism,increase the level of thyroid-associated hormone and insulin-like growth factor,promote the growth and development of children,which is worthy of clinical application.

Objective To investigate the roles of neuroinflammation and pathology of peripheral nervous system in the pathogenesis of Parkinson's disease (PD).Methods Immunohistochemical staining was performed to examine the nodose ganglia (containing vagus nerve) and intestine of an autopsy case of PD.The neuroinflammation and morphological changes of vagus nerve and enteric nervous system were observed.Results The expressions and distributions of glial fibrillary acidic protein and ionized calcium binding adapter molecule 1,two typical glia cell biomarkers,were different in vagus nerve and intestinal mucosa.Tumor necrosis factor α and inducible nitric oxide synthase were expressed in intestinal mucosa and myenteric plexus of the PD patient.There was a strong inflammatory reaction in the myenteric plexus,which distributed diffusely.Conclusion Satellite glial cells and intestinal inflammatory response may play a role in the pathogenesis of PD.

Objective:To investigate the clinical effect of the modified vertical rectus abdominis myocutaneous flap in repairing the skin and soft tissue defect after abdominoperineal resection for rectal cancer.Methods:This study was a retrospective observational study. From June 2019 to July 2022, five male patients with low rectal cancer who were conformed to the inclusion criteria were admitted to the Department of Basic Surgery of Xiangya Hospital of Central South University, with ages ranging from 65 to 70 years and the sizes of the perianal skin ulcers ranging from 5 cm×4 cm to 11 cm×9 cm, and all of them underwent abdominoperineal resection. The secondary skin and soft tissue defects in the perineum with an area of 8 cm×6 cm-14 cm×12 cm (with the depth of pelvic floor dead space being 10-15 cm) were repaired intraoperatively with transplantation of modified vertical rectus abdominis myocutaneous flaps with the skin area being 9 cm×7 cm-16 cm×12 cm, the volume of the muscle being 18 cm×10 cm×5 cm-20 cm×12 cm×5 cm, and the vessel pedicle being 18-20 cm in length. During the operation, most of the anterior sheath of the rectus abdominis muscle was retained, the flap was transferred to the recipient area through the abdominal cavity, the remaining anterior sheaths of the rectus abdominis muscle on both sides of the donor area were repeatedly folded and sutured, the free edge of the transverse fascia of the abdomen was sutured with the anterior sheath of the rectus abdominis muscle, and the donor area skin was directly sutured. After the operation, the survival of the transplanted myocutaneous flap was observed. The occurrence of complications in the perineal recipient area was recorded within 2 weeks after the operation. The recovery of the perineal recipient area and the abdominal donor area was observed during follow-up, and the occurrence of complications in the donor area of the abdomen as well as the recurrence of tumors and metastasis were recorded.Results:All transplanted myocutaneous flaps in 5 patients survived after surgery. One patient had dehiscence of the incision in the perineal recipient area 2 days after surgery, which healed after 7 d with intermittent dressing changes and routine vacuum sealing drainage treatment. In the other 4 patients, no complications such as incisional rupture, incisional infection, or fat liquefaction occurred in the perineal recipient area within 2 weeks after surgery. Follow-up for 6-12 months after discharge showed that the skin of the perineal recipient area had good color, texture, and elasticity, and was not bloated in appearance; linear scars were left in the perineal recipient area and the abdominal donor area without obvious scar hyperplasia or hyperpigmentation; no complications such as incisional rupture, incisional infection, intestinal adhesion, intestinal obstruction, or weakening of the abdominal wall strength occurred in the abdominal donor area, and the abdominal appearance was good with no localized bulge or formation of abdominal hernia; there was no local recurrence of tumor or metastasis in any patient.Conclusions:The surgical approach of using the modified vertical rectus abdominis myocutaneous flap to repair the skin and soft tissue defects after abdominoperineal resection for rectal cancer is relatively simple in operation, can achieve good postoperative appearances of the donor and recipient areas with few complications, and is worthy of clinical promotion.

Parkinson's disease (PD) is a common neurodegenerative disorder caused by the loss of dopamine neurons in the substantia nigra and the formation of Lewy bodies, which are mainly composed of alpha-synuclein fibrils. Alpha-synuclein plays a vital role in the neuroinflammation mediated by the nucleotide-binding oligomerization domain-, leucine-rich repeat-, and pyrin domain-containing protein 3 (NLRP3) inflammasome in PD. A better understanding of the NLRP3 inflammasome-mediated neuroinflammation and the related mitochondrial impairment during PD progression may facilitate the development of promising therapies for PD. This review focuses on the molecular mechanisms underlying NLRP3 inflammasome activation, comprising priming and protein complex assembly, as well as the role of mitochondrial impairment and its subsequent inflammatory effects on the progression of neurodegeneration in PD. In addition, the therapeutic strategies targeting the NLRP3 inflammasome for PD treatment are discussed, including the inhibitors of NLRP3 inflammatory pathways, mitochondria-focused treatments, microRNAs, and other therapeutic compounds.
Humans ; Parkinson Disease/complications* ; alpha-Synuclein ; Inflammasomes ; NLR Family, Pyrin Domain-Containing 3 Protein ; Neuroinflammatory Diseases ; Mitochondria

Scrotal angiokeratoma(Fordyce angiokeratoma)is often seen in elderly men presenting nodular hemorrhage. In this study, 8 cases of scrotal angiokeratoma were treated with 980nm diode laser. All the operations were successfully completed under local anesthesia without obvious complications and local symptoms improved. The results showed that 980nm diode laser is an alternative operation for the treatment of scrotal angiokeratoma.