Osteoporosis is a prevalent skeletal condition characterized by reduced bone mass and strength, leading to increased fragility. Buqi-Tongluo (BQTL) decoction, a traditional Chinese medicine (TCM) prescription, has yet to be fully evaluated for its potential in treating bone diseases such as osteoporosis. To investigate the mechanism by which BQTL decoction inhibits osteoclast differentiation in vitro and validate these findings through in vivo experiments. We employed MTS assays to assess the potential proliferative or toxic effects of BQTL on bone marrow macrophages (BMMs) at various concentrations. TRAcP experiments were conducted to examine BQTL's impact on osteoclast differentiation. RT-PCR and Western blot analyses were utilized to evaluate the relative expression levels of osteoclast-specific genes and proteins under BQTL stimulation. Finally, in vivo experiments were performed using an osteoporosis model to further validate the in vitro findings. This study revealed that BQTL suppressed receptor activator of NF-κB ligand (RANKL)-induced osteoclastogenesis and osteoclast resorption activity in vitro in a dose-dependent manner without observable cytotoxicity. The inhibitory effects of BQTL on osteoclast formation and function were attributed to the downregulation of NFATc1 and c-fos activity, primarily through attenuation of the MAPK, NF-κB, and Calcineurin signaling pathways. BQTL's inhibitory capacity was further examined in vivo using an ovariectomized (OVX) rat model, demonstrating a strong protective effect against bone loss. BQTL may serve as an effective therapeutic TCM for the treatment of postmenopausal osteoporosis and the alleviation of bone loss induced by estrogen deficiency and related conditions.
Animals ; NFATC Transcription Factors/genetics* ; Drugs, Chinese Herbal/pharmacology* ; Ovariectomy ; Osteoclasts/metabolism* ; Female ; Osteogenesis/drug effects* ; Rats, Sprague-Dawley ; Rats ; NF-kappa B/genetics* ; Osteoporosis/genetics* ; Signal Transduction/drug effects* ; Bone Resorption/genetics* ; Cell Differentiation/drug effects* ; Humans ; RANK Ligand/metabolism* ; Mitogen-Activated Protein Kinases/genetics* ; Transcription Factors

In this study, in vitro cytotoxicity of carboxymethyl chitosan-rhein conjugate(CR conjugate)and paclitaxel-loaded carboxymethyl chitosan-rhein polymeric micelles(PTX/CR PMs)was evaluated by MTT method in MCF-7 cells. The results showed that CR conjugate displayed good security; PTX/CR PMs in 24 h showed better antitumor activity than Taxol® . Environment-responsive fluorescent probe P4 was used to determine the cellular uptake of PTX/CR PMs in MCF-7 cells. The results also showed that P4 and PTX co-loaded carboxymethyl chitosan-rhein polymeric micelles ［(P4+PTX)/CR PMs］ could be taken up by MCF-7 cells. There was no difference between(P4+PTX)/CR PMs group and(P4+PTX)/CR PMs with verapamil group, suggesting that CR PMs could protect fluorescent probe and/or drugs in their cores avoiding efflux by P-glycoprotein. These results will contribute to in vivo study of CR conjugate and PTX/CR PMs in the future.

Objective To investigate the regularity and predictors of weight loss during concurrent chemo-radiotherapy for nasopharyngeal carcinoma patients. Methods A retrospective study was carried out on 82 patients with nasopharyngeal carcinoma ,with average age of 45 years ,who received concurrent chemoradiotherapy at the Affiliated Cancer Hospital of Guangzhou Medical University. Paired t-test was used to analyze the weight loss pattern during chemoradiotherapy. The Spearman′s rank correlation analysis was used to investigate the correlation between high weight loss(HWL)and the clinical factors. Results The paired t-test showed that the weight of the patients decreased continuously every week during chemoradiotherapy(P < 0.05),and the value dropped most significantly during the 4th week. Among these 82 patients,32 were determined as HWL(weight loss > 10%). Spearman′s rank correlation analysis showed that HWL was associated with older age,female,higher T and TNM stage,3D-CRT technology,the higher dose of radiotherapy in nasopharynx and in neck lymph node,and decreases of leukopenia and hemoglobin over degree Ⅱ(P<0.05). Conlousions Weight may decrease continuously every week during chemoradiotherapy for nasopharyngeal carcinoma patients. Age,gender,T stage,TNM stage,radio-therapy dose,and leukopenia and hemoglobin decreases may be predictors for weight loss degree during concurrent chemoradiotherapy for nasopharyngeal carcinoma patients.

Objective To identify the role of miR-124 in regulating the radiosensitivity and the epithelial-mesenchymal transition (EMT) of nasopharyngeal carcinoma (NPC). Methods Transient transfection of cells with miR-124 mimic or inhibitor was performed and wound-healing assay was used to investigate the role of miR-124 in the EMT of NPC. The apoptosis affected by miR-124 was also measured after irradiation , followed by investigating the cell proliferation by EdU assay. Finally , proteins of Akt and ERK associated with EMT and radiosensitivity, were measured by western blot. Results The migration index from NPC cell line indicated that miR-124 repressed the EMT. The results from caspase-3 activity assay showed that caspase-3 activity after irradiation significantly increased in miR-124 mimic group compared with the control group (P < 0.01). It was also confirmed that irradiation led to a higher percentage of apoptosis in miR-124 group compared with the control group in NPC cells. Cell proliferation after irradiation was significantly decreased in MiR-124 group as compared with control group . MiR-124 inhibited the protein expression of p-Akt . Conclusion MiR-124 may repress the EMT and decrease radio-resistance of NPC via p-Akt signaling pathway , which may provide a new insight into radio-resistance in NPC.

Objective To investigate the present status of early diagnosis and misdiagnosis of nasopharyngeal carcinoma (NPC). Methods A total of 50 patients diagnosed as NPC were recruited at the department of nasopharyngeal carcinoma in our hospital. The time from that patients felt discomfort to be diagnosed of NPC was determined bydifferent symptoms and signs. Furthermore , the status of misdiagnosis were also investigated. The spearmans rank correlation was used to analyze the correlation between the cancer stage and the time of confirmeddiagnosis , and the relationship between the rate of misdiagnosed and the rank of the hospital they visited. The χ2 test was then used to analyze the cancer stage with the time when they were diagnosed. Results Results indicated that the time when patients were diagnosed significantly correlated with pTNM stage (P < 0.05). Patients diagnosed in one month were most at stage Ⅱ, diagnosed in six months were stage III, and diagnosed after twelve months were stage Ⅳ(P < 0.05). Furthermore, we found that the rate of misdiagnosis of NPC was 12%. Moreover,the misdiagnosed rate was associated with the rank of the hospital patients visited. Discussion In conclusion , the present status of the early diagnosis of NPC is not optimistic. Most of the patients with NPC were misdiagnosed in basic medical institutions , especially in town or village health center. Thus, it is important to popularize the health knowledge about the secondary prevention of NPC and train the doctor in basic medical institutions.

Objective The radiotherapy resistance is one of important causes for nasopharyngeal carcinoma(NPC) treatment failure.Junctional adhesion molecule A(JAMA)is closely correlated with the tumor poor prognosis.Thus this experiment is to in vestigate the relationship between JAMA expression and the radiosensitivity of NPC.Methods To overexpress or interfere the JAMA expression in CNE2 and HONE1 cell lines.Then different doses of X-ray were adopted to conduct irradiation.The cell clone formation capacity and cellular apoptosis change were detected after 24 h.The role of JAMA in the NPC radiotherapy was understand.The related signal pathway protein in cell lines with different JAMA expression was detected by Western blot.Results The cell lines with low JAMA expression were more sensitive to radiotherapy:After low JAMA expression,the D0 value in the CNE2 cell line was decreased from 3.26 ±0.78 to 1.92 ± 0.23;the Dq value was decreased from 46.51 ± 4.27 to 32.12 ± 3.19.The radio therapy induced apoptosis was significantly increased in the cell lines with low JAMA expression,after low JAMA expressing,thcellular apoptosis was elevated from 6.9 ％ ± 0.9 ％ to 13.7 ％ ± 1.3 ％;the HONE1 cellular apoptosis was elevated from 6.5 ％ + 1.1 ％ to 12.3 ％ ± 1.7％;JAMA overexpression cell lines were significantly decreased.The preliminary mechanism research results showed that JAMA played the effect via Akt signal pathway.Conclusion This research results verifiy that JAMA expression level is closely correlated with the radiosensitivity of NPC cell line:JAMA can increase the radiotherapy resistance of NPC cell lines,which provides a new feasible research direction for NPC enhancing radiosensitivity.

This study aimed at investigating the correlations between antidepressive effect of valproate and improvements of NET and 5-HTT expression in depressive rats leaded by chronic mild unpredicted stress(CUMS)with solitary condition. Sixty male SD rats were divided into normal group(NG), model group(MG), valproate treated-normal group(VNG), and valproate treated-model group(VMG), randomly. The changes of depressive behaviors were evaluated by the open-field test and force swimming test. The contents of MDA, activities of SOD and CAT in serum, the mRNA and protein expression of NET, 5-HTT in hippocampus were determined by biochemical methods, Real-time PCR and Western Blot, respectively. Results showed that CUMS can significantly decrease the activities in open-field test, SOD and CAT activities in serum, expression of 5-HTT in hippocampus, and obviously increase the immobility time in force swimming test, the level of MDA and expression of NET. The treatment of valproate obviously improved the changes induced by CUMS. However, the treatment of valproate had no significant influences on behaviors of NG rats. So, it revealed that improvements of the mRNA and protein expression of NET, 5-HTT may be involved in the antidepressive effect of valproate.

Aim To investigate whether chronically un-predictable stress (CUS)-induced depression-like be-haviors of rats is associated with the variant expression of tryptophan hydroxylase (TPH)and tyrosine hydrox-ylase (TH).Methods 30 male SD rats were ran-domly divided into depression model group(MG)and control group (CG),the former was established using CUS plus solitary condition for 28 d,whereas the latter was fed normally as five rats per cage without CUS. The open field test(OFT)and the sucrose preference test were used to evaluate depressive behaviors.Both mRNA and protein expressions of TPH and TH in hip-pocampus and forebrain cortex were determined by re-al-time fluorescent quantitative PCR and western blot (WB),respectively.Results MG rats showed obvi-ous depressive behaviors with much lower locomotive activity and sucrose preference than CG.Meanwhile, the mRNA and protein expressions of TPH and TH also significantly decreased in MG rats,compared with CG rats.Conclusion The depression behaviors of rats in-duced by CUS may be associated with down-regulation of TPH and TH expression.

Aim To investigate if anti-depressive effect of venlafaxine was associated with improving oxi-dative stress and expression of hippocampus NET and 5-HTT in rat model induced by CUS. MethodsEighty SD male rats were randomly divided into four groups:model group(MG),normal group(NG), ven-lafaxine-treated normal group ( VNG ) , and venlafax-ine-treated model group ( VMG) . VNG and VMG were given venlafaxine (23. 4 mg·kg-1 ·d-1 ) once daily;NG and MG were given the same volume solvent. Soli-tary condition with chronic unpredicted stress ( CUS ) was taken to establish rat depression model. The force swimming test was used to evaluate the behavior chan-ges of experimental rats. The malondialdehyde ( MDA) level and activity of superoxide dismutase ( SOD ) in serum were determined by biochemical methods. The mRNA and protein expressions of NET and 5-HTT in
hippocampus were determined by Real-Time Reverse transcription polymerase chain reaction ( real-time RT-PCR) and Western blot ( WB) , respectively. Results Compared with NG rats, obviously increasing immo-bile time of rats in force swimming test and serum MDA level, as well as significantly decreasing SOD activity in serum was observed with clearly decreasing 5-HTT expression and elevating NET expression in hippocam-pus of MG rats. The treatment of venlafaxine distinctly suppressed changes above from CUS-induced rats. However, significant changes failed to be found in VNG rats. Conclusion The anti-depressive effect of venlafaxine may at least partly involve in improving ox-idative stress/anti-oxidative stress balance and revers-ing abnormal expression of NET and 5-HTT.

Aim To study the effect of the total saponins of Rubus parvifolius(TSRP)on the intracellular free calcium levels in ischemic hippocampus neurons.Methods The cytosolic free calcium concentration in ischemic hippocampal neurons was measured by using Ca2+-sensitive fluorescent probe fluo-2/AM with a laser scanning confocal microscope.Results Application of TSRP inhibited free intracellular calcium in ischemic hippocampus neurons in a concentration-dependent manner.Conclusions These results suggest that TSRP might protect hippocampus neurons via attenuating calcium overload induced by ischemia.