BACKGROUND:Overweight or obesity is the most significant risk factors for knee osteoarthritis.Time-restricted diet shows an effective effect in preventing overweight or obesity.Whether infrapatellar fat pad,an important component of the knee joint,is affected by time-restricted diet and thus influences knee osteoarthritis remains unclear. OBJECTIVE:To employ a high-fat diet-induced rat model to investigate the effect of weight loss by time-restricted diet on infrapatellar fat pad,thereby providing evidence for early prevention and treatment of obesity-related knee osteoarthritis. METHODS:Fifteen male Sprague-Dawley rats were randomly divided into three groups(n=5 per group).Rats in the control group were allowed to eat at will for 24 hours and were fed the defined control diet(12%fat);rats in the high-fat diet group were allowed to eat at will for 24 hours and were fed the high-fat diet(45%fat);and rats in the high-fat diet with time-restricted diet group were fed the high-fat diet only from 9:00(2 hours after the light)to 17:00(2 hours before the dark).After 8 weeks of feeding,mDixon-Quant sequence was used to assess proton density fat fraction in the infrapatellar fat pad and subcutaneous adipose tissue in the right inguinal region.ELISA was used to quantify differences adipokine.Sirius red staining was used to evaluate changes in fibrosis of the infrapatellar fat pad.The expressions of uncoupling protein-1 in the infrapatellar fat pad and subcutaneous adipose tissue in the right inguinal region as well as leptin,adiponectin and tumor necrosis factor-α in the infrapatellar fat pad were detected by immunohistochemistry staining. RESULTS AND CONCLUSION:After 8 weeks of feeding,compared with the high-fat diet group,the body mass of rats in the control group(P=0.036)and the high-fat diet with time-restricted diet group(P=0.003)was significantly reduced.The proton density fat fraction in the infrapatellar fat pad in the high-fat diet group was significantly higher than that in the control group(P<0.001)and the high-fat diet with time-restricted diet group(P=0.004),while there was no significant difference in the proton density fat fraction of the subcutaneous adipose tissue among the three groups.The serum leptin levels of rats in the high-fat diet group were significantly higher than those in the control group(P=0.030)and the high-fat diet with time-restricted diet group(P=0.018).Compared with the high-fat diet group,the infrapatellar fat pad fibrosis characterized by Sirius red staining in the control group(P<0.001)and the time-restricted diet group(P=0.003)was significantly decreased.The expression of leptin in IFP of the high-fat diet group was significantly higher than that of the control group(P<0.001)and the high-fat diet with time-restricted diet group(P<0.001).The expression of adiponectin in the infrapatellar fat pad of the high-fat diet group was significantly lower than that of the control group(P=0.004)and the high-fat diet with time-restricted feeding group(P=0.048).However,there was no positive expression of uncoupling protein-1 in the infrapatellar fat pad and subcutaneous adipose tissue of all the three groups,and no positive expression of tumor necrosis factor-α in the infrapatellar fat pad.To conclude,time-restricted diet could retard the fibrosis of the infrapatellar fat pad,reduce the proton density fat fraction of the infrapatellar fat pad,and affect the level of adipokine in serum and infrapatellar fat pad.Time-restricted diet may become a simple and effective option for the treatment and prevention of obesity-related knee osteoarthritis.

Objective:To investigate the effect of time-restricted feeding (TRF) on bone marrow fat of proximal femur in obese rats induced by high-fat diet (HFD) using proton density fat fraction (PDFF).Methods:Totally 30 male Sprague-Dawley rats were stratified and randomly sampled into 6 subgroups according to body weight, with 5 rats each. Then 2 subgroups were combined into one group, and there were totally 3 groups. The rats in the control group were fed with normal diet, and the rats could eat as much as they wanted for 24 h; the rats in the HFD group were fed with high-fat diet, and the rats could eat as much as they wanted for 24 h; the rats in the HFD+TRF group were fed with high-fat diet only between 9 AM (2 h after light) and 17 PM. One subgroup of rats from each group was examined with MRI on the femur on day 28 of the experiment, and the other subgroup from each group was examined on day 56 to measure the bone marrow PDFF of proximal femur based on mDixon-Quant quantitative sequence images. The rats were executed at the end of the scanning period, and blood samples were collected to measure serum levels of leptin. One-way ANOVA or Kruskal-Wallis H test was used to compare the differences in body weight, PDFF, and serum levels of leptin among 3 groups. The LSD- t test was used for multiple comparisons. Results:On day 28 of the experiment, the differences in body weight, PDFF, and serum leptin among the 3 groups of rats were not statistically significant ( P>0.05). On day 56, the bone marrow PDFF of proximal femur of the rats in the control group, HFD group, and HFD+TRF group were (7.2±1.4)%, (9.7±2.4)%, and (11.2±3.6)%, respectively. The differences in body weight, PDFF, and serum levels of leptin among the 3 groups of rats were statistically significant ( F=6.95, P=0.010, F=5.98, P=0.007, F=4.54, P=0.034). The results of multiple comparisons showed that the body weight in the HFD group was higher than those in the control group (LSD- t=52.96, P=0.036) and the HFD+TRF group (LSD- t=82.74, P=0.003). The values of bone marrow PDFF of proximal femur in the HFD+TRF group was higher than that in the control group (LSD- t=4.01, P=0.012). The serum levels of leptin in the HFD group were higher than those in the control group (LSD- t=1.45, P=0.030) and the HFD+TRF group (LSD- t=1.62, P=0.018). Conclusion:TRF induces an increase in the values of bone marrow PDFF of proximal femur in conjunction with weight loss in obese rats induced by HFD, and the increase in bone marrow fat may be related to the decrease in serum leptin.

Rats ; Animals ; Adipose Tissue, Brown/physiology* ; Adipose Tissue

Objective:To analyze clinical manifestations, histopathological and pathogenic fungus characteristics as well as treatment of 3 cases of disseminated cutaneous alternariosis caused by Alternaria. Methods:Clinical data were collected from 3 cases of disseminated cutaneous alternariosis caused by Alternaria, who were diagnosed in Department of Dermatology, Xijing Hospital from 2019 to 2021, and clinical and histopathological features, fungal culture, strain identification and treatment results were retrospectively analyzed. Results:The 3 patients were aged 55, 41 and 46 years respectively, including 1 male and 2 females. Two patients were previously diagnosed with nephrotic syndrome and 1 with systemic lupus erythematosus. All the patients had a history of taking glucocorticoids and tacrolimus for different durations, and experienced chronic infections. Histopathological examination with hematoxylin and eosin (HE) staining showed double-contour thick-walled spores and knot-shaped thick-walled septal hyphae, but no melanin in skin lesions. Sequencing of the fungal internal transcribed spacer region confirmed that 2 cases were infected with Alternaria alternate, and 1 with Alternaria infectoria. Fungal culture at different temperatures showed that the growth ability of Alternaria markedly decreased at the temperature over 35 ℃. To treat these patients, the dose of tacrolimus was reduced to less than 1/3 of the standard dose, or tacrolimus was switched to other immunosuppressants, and systemic antifungal therapy was also given at the same time. After 7-month treatment, good clinical outcomes were achieved in the 3 patients. Conclusion:Disseminated cutaneous alternariosis is characterized by bilateral hematogenous dissemination and lymphatic distribution in unilateral limbs, and the skin lesions are characterized by verrucous plaques covered with scabs, nodules and/or sinuses.

Objective:To understand the current status of nurses' awareness and needs for the Employee Assistance Program (EAP), as to provide basis for the hospital to formulate EAP.Methods:Using the purpose sampling method, 352 ICU nurses from the Nanjing Drum Tower Hospital Affiliated of Nanjing University Medical School were selected as the research subjects from December 2019 to January 2020. Finally, 350 valid questionnaires were obtained. Use the EAP demand questionnaire to investigate the ICU nurses' awareness and demand for EAP.Results:Among the 350 nurses in the intensive unit, 214 cases (61.14%) had never understood EAP, only 8 (2.29%) had understood and experienced the EAP, and 332 cases (94.86%) believed that the hospital′s implementation of EAP was a benefit provided to nurses. The top two service requirements were: "Relief of work pressure" and "mental health consultation", and hope to receive mixed-mode EAP services; in ICU and CCU, nurses hope EAP can help resolve work-family conflicts; in EICU, they need to provide Specific job skills training services.Conclusions:ICU nurses have insufficient cognition of EAP and urgently need to obtain EAP implementation knowledge; at the same time, it is necessary to formulate an implementation plan that meets ICU nurses′ needs for EAP, that is, to relieve work pressure and carry out psychological counseling to ensure that EAP is launch successfully in ICU nurses.

OBJECTIVETo assess the effects of laparoscopic sleeve gastrectomy (LSG) on insulin secretion mode and metabolism of glucose and lipid in morbidly obese patients.

METHODSClinical data of 65 morbidly obese patients [body mass index (BMI) ≥30 kg/m] undergoing LSG at Shanghai 10th People's Hospital from August 2012 to December 2016 were retrospectively analyzed. According to the result of OGTT, these obese patients were divided into three groups: normal glucose tolerance (NGT, 23 cases), impaired glucose tolerance (IGT, 22 cases) and type 2 diabetes mellitus (DM, 20 cases) groups. Twenty-two healthy people [BMI (23.1±1.4) kg/m] were used as control group. The anthropometries parameters [weight, BMI, waist circumference, body fat percentage, excess weight loss(%EWL)], glucose metabolic indices [fasting plasma glucose (FPG), fasting insulin (FINS), glycosylated hemoglobin (HbA1c), homeostasis model assessment-insulin resistance index (HOMA-IR)], lipid profile (TC, TG, HDL-C, LDL-C) and inflammatory factor (UA, TNF-α) of 3 groups were detected before operation and at postoperative 1-, 3-, 6-month. These variables were analyzed among morbidly obese groups before and after surgery and compared to control group. Clinical registration number of this study was ChiCTROCSl2002381.

RESULTSBody weight, waist circumference and BMI of morbidly obese patients all decreased at postoperative 1-, 3-, 6-month. Postoperative %EWL increased obviously to (71.5±24.7)% with the highest range in DM group. Percentage of successful weight loss (%EWL>50%) in NGT, IGT and DM groups was 63.6%, 83.9% and 90.0% at postoperative 6-month respectively, and DM group was also the highest. At postoperative 6-month, HbA1c of 3 morbidly obese groups became normal; FPG and postprandial 2-hour glucose of IGT and DM group decreased to normal level; insulin level of 3 morbidly obese groups decreased obviously compared to pre-operation (all P<0.05), especially FINS and postprandial 2-hour insulin became normal without significant difference of control group (P>0.05), while postprandial 30-minute and 60-minute insulin levels in 3 groups were still higher as compared to control group. The insulin secretion curves of morbidly obese groups showed hyperinsulinemia before surgery. The peak of insulin secretion curve in IGT and DM group moved back to postprandial 120-minute before operation, and returned to 60-minute after operation, with basic normal rhythm of secretion curve. Preoperative HOMA-IR in all 3 morbidly obese groups was higher than that in control group (all P<0.05) and remarkably lower at postoperative 6-month compared to pre-operation(P<0.05). In 3 morbidly obese groups after operation, TG decreased, HDL-C increased, UA and TNF-α decreased significantly compared to before operation (all P<0.05). At postoperative 6-month, the HOMA-IR of DM group was positively correlated with BMI (r=0.236, P=0.004) and TNF-α (r=0.228, P=0.033), and was not correlated with HDL-C(P>0.05).

CONCLUSIONSLSG can effectively ameliorate hyperinsulinemia and insulin secretion curve, and improve metabolic disorder and insulin resistance of different stage in obesity patients with glucose metabolic disorder. Insulin resistance is correlated with body weight and inflammatory factors.

Objective To assess the variation of sexual hormone and mechanisms of low testosterone in young male obesity with acanthosis nigricans. Methods Retrospective analysis was performed in 125 male obesity patients [ body mass index( BMI)≥28 kg/m2 ] . According to their clinical characteristics, they were divided into two groups including obesity without acanthosis nigricans(OB group, n=62) and obesity with acanthosis nigricans(AN group, n=63). 60 normal weight men were also recruited as a control group. Body fat and body weight were measured. Blood insulin, lipid profile, sex hormones levels, and inflammation factors were measured. Parameters of each group were compared and the correlations between total testosterone level and other index were analyzed. Results All the male obesities have the significant lower total testosterone levels than those of control group(P>0. 05), and those in AN group were lower than those in OB group(P>0. 05). The BMI and body fat in OB group and AN group were both significantly higher than those in control group(P>0. 05). The fasting insulin levels in all obese men were significantly higher than those in control group(P>0. 05), highest in AN group. Triglycerides(TG) in both OB and AN group were higher than those in controls, and not significant between later 2 groups. But high-density lipoprotein-cholesterol ( HDL-C) in the two groups were significantly lower than control, which in AN group were significantly lower than OB group. Total testosterone levels in AN group were negatively correlated with weight, waist circumference, hip circumference, fasting insulin, and homeostasis model assessment for insulin resistance ( HOMA-IR ) , and also negatively correlated with inflammation factors including C-reactive protein ( CRP ) , erythrocyte sedimentation rate ( ESR) , tumor necrosis factor-α( TNF-α) , and uric acid. However, total testosterone levels in AN group were not correlated with lipid metabolism index. Conclusion Young male obesity with acanthosis are associated with secondary hypogonadism. Hyperinsulinemia, insulin resistance, and inflammatory factors are risk factors for the occurrence of this secondary male hypogonadism.

Objective To investigate the distribution of gut microbiota in obese patients with or without acanthosis nigricans .Methods Totally 131 obese patients and 25 healthy participants were divided into three groups:the obesity with acanthosis nigricans (AN) group (n=59), the simple obesity (OB) group (n=79), and the control (CON) group (n=25).The fresh stool samples were collected , and the clinical and biochemistry markers were measured .Pyrosequencing technology was performed based on the 16s rRNA of fecal samples to identify and analyze the distribution pattern of gut microbiota in each group .Results The AN group had signifi-cantly higher body mass index [ (37.45 ±5.12) kg/m2 vs.(33.34 ±2.54) kg/m2 vs. (20.35 ±1.68) kg/m2, P=0.045, P<0.001], insulin [32.77 (25.18) mU/L vs.20.73 (9.30) mU/L vs.8.70 (6.18) mU/L, P<0.001, P<0.001], insulin resistance [7.78 (6.87) vs.4.71 (2.88) vs.1.81 (1.40), P<0.001, P<0.001], and interleukin (IL) -6 [ (3.64 ±2.23) ng/L vs.(2.71 ±0.78) ng/L vs.(2.17 ±0.86) ng/L, P=0.040, P=0.009] levels than OB and CON groups compared with OB and CON groups , AN group had sig-nificantly decreased diversity of bacterial flora ( P=0.015 , P=0.001 ) , while no significant difference was observed in the abundance of bacterial flora .At the phylum level , the composition of flora among these three groups was similar, mainly including bacteroidetes , firmicutes, proteobacteria, and actinomycetes.Although the proportions of main bacteria flora were different , the difference was not statistically significant .At the genus level, the bacteria flora in AN and OB groups were primarily composed of Bacteroides, Megamonas, Faecalibac-terium and Escherichia-Shigella.In addition, compared to OB and CON groups , AN group had significantly lower proportion of Ruminococcus ( P=0.023 , P=0.043 , respectively ) and higher proportion of Veillonella (P=0.048, P=0.043, respectively).Furthermore, the proportion of Weissella was higher in AN and OB groups than in CON group ( P=0.045 , P=0.025 ) .Conclusion Obese patients with AN have more severe in-sulin resistance and inflammation status than those with simple obesity , and the distribution feature of gut micro-biota also differ between these two patient populations .

OBJECTIVETo investigate the effect of laparoscopic sleeve gastrectomy(LSG) on sex hormone in male patients with severe obesity.

METHODSRetrospective analysis was performed in 31 male patient with severe obese [body mass index(BMI) ≥28 kg/m, obesity group] who underwent LSG in Shanghai Tenth People's Hospital of Tongji University from December 2012 to May 2016. The anthropometric parameters(weight, BMI, waist circumference, hip circumference, waist-hip ratio, body fat percentage), glucose metabolic indices [fasting plasma glucose(FPG), fasting insulin (FINS), glycosylated hemoglobin (HbA1c), homeostasis model assessment-insulin resistance index(HOMA-IR)], and sex hormone parameters [estradiol(E2), total testosterone (TT), follicle-stimulating hormone (FSH) and luteinizing hormone (LH)] were collected preoperatively and 1, 3, 6 months postoperatively. In addition, 31 healthy male volunteers with normal BMI were consecutively recruited in this study as control group. The above-mentioned parameters were also determined in control group. Changes of these variables before and after surgery were analyzed. Pearson method was used to analyze the correlation of TT with anthropometric parameters and glucose metabolic indices before and after surgery.

RESULTSThe average age of patients in obesity and control group was (32.9±9.7) (18 to 56) years and (30.7±8.9) (18 to 49) years. Compared to the control group, obesity group had significantly higher anthropometric parameters and glucose metabolic indices before surgery (all P<0.05). In obesity group, the anthropometric and glucose metabolic indices significantly decreased at 1 to 6 months after surgery compared to those before surgery (all P<0.05). At 1 month after surgery, the anthropometric parameters and glucose metabolic indices in obesity group were significantly higher than those in control group (all P<0.05). At 3, and 6 months after surgery, there were no significant differences in glucose metabolic indices between obesity and control group (all P>0.05), while the anthropometric parameters in obesity group were still significantly higher than those in control group(all P<0.05). The sex hormone parameters in control and obesity group before surgery were as follows: E2: (100.2±23.5) pmol/L and (129.2±81.9) pmol/L; TT: (18.0±4.9) nmol/L and (8.4±4.5) nmol/L; FSH: (4.5±3.1) IU/L and (4.3±2.5) IU/L; LH: (4.4±1.7) IU/L and (5.3±2.6) IU/L. Compared to control group, the TT level of obese patients before surgery significantly decreased(P=0.000), while no significant differences were observed in the levels of E2, FSH, and LH(all P>0.05). The TT levels were significantly increased at 1, 3, 6 months after surgery[(13.1±7.0), (13.6±5.7), (21.0±19.3) nmol/L, respectively, all P<0.05] and the E2 level was significantly decreased at 6 months after surgery [(91.4±44.9) pmol/L, P<0.05], while no significant differences were observed at 1 and 3 months after surgery (all P>0.05). Furthermore, the FSH and LH levels did not exhibit significant change at 1, 3, and 6 months after surgery compared to those before surgery (all P>0.05). At 1 month after surgery, no significant correlations were examined in the change value of TT levels (▹TT) with the changes of BMI(▹BMI), FPG(▹FPG), FINS(▹FINS), HOMA-IR(▹HOMA-IR), and E2(▹E2) (all P>0.05). At 3 months after surgery, ▹TT was negatively correlated with ▹BMI (r=-0.441, P=0.015), ▹FINS (r=-0.375, P=0.041), and ▹HOMA-IR(r=-0.397, P=0.030), but not correlated with ▹FPG and ▹E2 (all P>0.05). At 6 months after surgery, ▹TT was negatively correlated with ▹BMI(r=-0.510, P=0.018) and ▹HOMA-IR (r=-0.435, P=0.049), but not correlated with ▹FPG, ▹FINS and ▹E2 (all P>0.05).

CONCLUSIONSMale severe obese patients are accompanied with abnormal sex hormone levels. LSG has a significant effect on weight loss and blood glucose improvement, and may ameliorate the sex hormone unbalance by improving the insulin resistance in men with severe obesity.

Adult ; Bariatric Surgery ; Blood Glucose ; physiology ; Body Mass Index ; Body Weights and Measures ; China ; Estradiol ; blood ; physiology ; Fasting ; blood ; Follicle Stimulating Hormone ; blood ; physiology ; Follow-Up Studies ; Gastrectomy ; Glycated Hemoglobin A ; physiology ; Humans ; Insulin ; blood ; physiology ; Insulin Resistance ; physiology ; Luteinizing Hormone ; blood ; physiology ; Male ; Obesity, Morbid ; surgery ; Retrospective Studies ; Testosterone ; blood ; physiology ; Treatment Outcome ; Weight Loss ; physiology

Objective To monitor the autophagy in osteosarcoma cells by constructing three rLC3B fusion expression vectors,respectively.Methods Rat LC3B gene sequence was amplified by PCR and cloned into pEGFP-C 1 and pmCherry-C1 to construct the fusion expression vector of pEGFP-rLC3B and pmCherry-rLC3B.Subsequently,the EGFP-rLC3B sequence was obtained by PCR with the pEGFP-rLC3B as a template,and cloned into pmCherry-C 1,so the pmCherry-EGFP-rLC3B fusion expression vector was constructed.Three plasmids were transfected into U-2OS cells,and the starvation or Rapamycin was adopted to induce autophagy or the chloroquine or Baf-A1 was used to inhibit autophagy,to verify the above plasmids' function in autophagy detection by laser scanning confocal microscopy.Western blot was used to detect the endogenous LC3B and exogenous EGFPrLC3B,pmCherry-rLC3B and mCherry-EGFP-rLC3B,and to verify the correct expression of exogenous rLC3B and their function of autophagy detection.Finally,cleaved free EGFP was detected by western blot to evaluate the level of autophagic degradation.Results Three fusion expression vectors were constructed successfully through sequencing and restriction enzyme digestion validation.The starvation or Rapamycin was adopted to induce autophagy or the chloroquine or Baf-A 1 was used to inhibit autophagy in transfected U-2OS cells.Clear autophagosomes and autolysosomes were observed by laser scanning confocal microscopy.Endogenous LC3B and exogenous EGFP-rLC3B,pmCherry-rLC3B and mCherry-EGFP-rLC3B were detected through western blot.Finally,western blot verified that the expression of cleaved free EGFP was significantly up-regulated with the increase of starvation time.12 h group increased 1.05 times than the control group and 24 h group increased 1.56 times,showing that the levels of autophagic degradation increased.Conclusion EGFP-rLC3B can be used to detect autophagosome and evaluate the level of autophagic degradation.mCherry-rLC3B can be used to detect autophagosome and autolysosome,but can't distinguish autophagosome from autolysosome.The pmCherry-EGFP-rLC3B has an advantage in the detection of autophagic flux which can distinguish autophagosome from autolysosome.