Bioethnopharmaceutical analytical pharmacology (BAP) means to study Chinese herbal compound formula (CHCF) from the aspacts of in vivo and in vitro efficacy,pharmacodynamics,quality control and plant chemistry,guided by the CHCF absorbed bioactive compounds (ABC) analyses.The form of BAP is performed by comparing ABC efficacy with mother formula efficacy.Meanwhile,it must follow the principle which the ABC dose should be equal to the mother formula content or the blood drug concentration.In this study,the hypothesis was put forward to clarify the thoughts,assumptions and expected results,which uncovered the multiple cardioprotective mechanism of Dangui-Buxue Tang (DBT) for ischemic heart disease.BAP is expected to guide the development of further experiments for providing a better thought for the research over CHCF.

OBJECTIVETo evaluate the influence of eugenol-containing and resin-containing endodontic sealers on the bond strength of fiber posts using different strategies of root canal irrigation.

METHODSForty-eight mandibular premolars were endodontically treated. The specimens were randomly assigned into two groups according to different endodontic sealers. Group A used Endofil (eugenol-containing endodontic sealer), and group B used AH-plus (resin-containing endodontic sealer). After post space preparation, each group was randomly assigned into three subgroups according to the strategies of root canal irrigation (eight premolars in each subgroup). Group Al and B1: 0.9%NaCl irrigation; Group A2 and B2: 17% ethylene diamine tetraacetic acid (EDTA)+5.25%NaClO+0.9%NaCl irrigation; Group A3 and B3: ultrasonic agitation associated with 1 7%EDTA+5.25%NaClO+0.9%NaCl. One week after the cementation of fiber posts using RelyX™ Unicem, a push-out test was performed to measure the bond strength of the posts. The microstructure of the root canal surface was examined under scanning electron microscope (SEM).

RESULTSThe bond strengths of the six groups were as follows: Al (7.96±2.23) MPa, A2 (9.95±2.89) MPa, A3 (18.88±3.69) MPa, B1 (11.41±3.71) MPa, B2 (14.00±4.04) MPa, and B3 (19.14±3.27) MPa. Statistical analysis revealed a significant interaction between the different endodontic sealers and the strategies of root canal irrigation (P<0.05). Lower bond strength was found in group Al but not in group BI (P<0.05), and the same result was revealed when comparing group A2 and B2. No significant difference was observed between group A3 and B3 (P>0.05). SEM showed that the root canal in group A3 and B3 achieved the cleanest surface with nearly all dentine tubules opened.

CONCLUSIONThe eugenol-containing endodontic sealer can impair the bond strength of fiber posts compared with the resin-containing sealer when the root canal is irrigated by 0.9% NaCl or 17%EDTA+5.25%NaClO+0.9%NaC. No difference was observed between the two sealers when using 17%EDTA+5.25% NaCIO+0.9%NaCl combined with ultrasonic irrigation.

Bicuspid ; Cementation ; Dental Bonding ; Dental Pulp Cavity ; Dental Stress Analysis ; Dentin ; Humans ; Post and Core Technique ; Root Canal Filling Materials ; Root Canal Irrigants ; Root Canal Therapy

OBJECTIVETo evaluate the effect of different auxiliary resistance forms on the resistance and marginal fitness of complete crowns for short molar preparations.

METHODSA total of 70 Nissin resin teeth were prepared with 20° total occlusal convergence, 2.5 mm of occlusocervical height, and a shallow finish line on a milling machine. The milled preparations were then randomly assigned to 7 groups of 10. The first group was used as the control group. A total of 30 dies were modified by preparing interproximal grooves with angles of 0°, 6°, and 20° centered on the mesial and distal surfaces of the dies. The rest of the teeth were prepared with occlusal holes in the center of the occlusal surface milled with the same burs to form 0°, 6°, and 20° holes. Cobalt-chromium copings were fabricated for all specimens. The marginal gap of specific points on the axial surface was measured before and after cementation. The resistance of each specimen was evaluated by applying an external force at an angle of 45° to the long axis of the die by using a universal testing machine in a lingual to buccal direction. The maximum force applied before crown dislodgement was measured. Data were analyzed using the SAS 9.2 software.

RESULTSThe results showed that the 0° groove, 0° hole, and 6° hole were effective in improving the resistance of the complete crowns (P<0.05). The 0° groove, 6° groove, 0° hole, 6° hole, and 20° hole had significant difference with the control group in terms of marginal discrepancies (P<0.05).

CONCLUSIONAuxiliary resistance forms with less degree indicate greater resistance force but worse marginal fitness. In clinical practice, if the resistance of a preparation is enough, the auxiliary resistance forms should be avoided from being used.

Cementation ; Crowns ; Dental Prosthesis Design ; Dental Prosthesis Retention ; Humans ; Molar ; Tooth Crown

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 .

Objective To investigate the clinical efficacy of Shenmai injection (SMI) combined with heart transplantation of autologous bone marrow stem cells in the treatment of refractory heart failure(RHF). Methods Two hundred patients with RHF were selected from the Emergency Department of the First Affiliated Hospital of Zhengzhou University and the Cardiology Department of the First Affiliated Hospital of Henan College of Traditional Chinese Medicine. They were randomly divided into control group and treatment 1,2 and 3 groups(each 50 cases). In the control group,heart failure standard treatment was given;group 1 received a standard treatment for heart failure combined with autologous bone marrow stem cell heart transplantation;group 2 received a standard treatment for heart failure with SMI;group 3 received a standard treatment for heart failure combined with autologous bone marrow stem cell heart transplantation and SMI. The mean follow-up was 24 months. The prognosis,readmission rate,clinical efficacy,cardiac function and the change in levels of B-type natriuretic peptide(BNP)of patients in each group were observed during the therapeutic course and observation period. Results During the course of treatment,there were 10 cases dead in the control group,4 in each group 1 and 2 respectively,and 3 in group 3. Readmission rates in group 1,2 and 3 were significantly lower than that in control group(38%,36%,24%vs. 48%, P<0.05 or P<0.01). The rates of total efficiency in group 1,2 and 3 were obviously higher than that in the control group(88%,86%,94%vs. 76%,all P<0.05). After treatment,the left ventricular ejection fraction(LVEF),left ventricular fractional shortening(FS)and left ventricular end-systolic diameter(LVESD)in three therapeutic groups were significantly higher than those before treatment,while the left ventricular end-diastolic diameter(LVEDD)and BNP level were significantly lower than those before treatment. All the above indexes in three therapeutic groups after treatment were much more remarkably improved than those in the control group during the same period,and the group 3 being the most significant〔LVEF:0.477±0.099 vs. 0.396±0.098,FS:(30.0±5.1)%vs.(26.8±7.5)%,LVESD (mm):40.6±9.1 vs. 45.8±9.4,LVEDD(mm):44.9±9.8 vs. 52.8±10.1,BNP(ng/L):515±400 vs. 1 875±400, all P<0.05〕. Conclusion SMI combined with heart transplantation of autologous bone marrow stem cells has obvious therapeutic effect for treatment of RHF.

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.

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.

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 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 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.