BACKGROUND: In the research, the recipe for kidney tonification is prepared into "topical form" and "oral form". According to the interrelationship of "acupoints-meridans and collaterals-internal organs-target organs", osteoporosis is treated by the approaches of point compress and oral application respectively. According to the traditional theory of Chinese medicine, it is to infer that the herbs bring the adjustment into play by"being attributive to kidney meridian", but,whether does the specialty of herbs on "targeting medication" happenpractically or not? Hormones in "hypothalamus-pituitary- target gland" system is assayed to further analyze estrogen receptor and androgen receptor of target organs,such as kidney,bone, uterus, thyroid gland and testis so as to observe the relativity between "tropism and receptor".DESIGN: Complete random design with the objects of osteoporosis rats, and the control experimental study with kidney tonification recipe administrate with different approaches.OBJECTIVE: To probe into the effect of kidney tonification recipe on pituitary-thyroid axis in experimental osteoporosis.SETTING: College of Traditional Chinese Medicine of Hebei Medical University.MATERIALS: The experiment was performed in Chinese-Western integrated Basic Experiment Room of Hebei Medical University from January 2000 to December 2004. Sixty SD healthy female rats of 3-month old were employed,weighted(300 ± 20) g. The experimental animals were provided by Hebei Experimental Animal Center. After bred routinely for 1 week in Experimental Room, the rats were randomized into 6 groups,namely normal control(10rats), pathological model group(10 rats), infusing group(10 rats) in which kidney tonification recipe was applied based on 0. 8 g/100 g body mass,acupoint paste on bladder meridian group (10 rats), acupoint paste on kidney meridian group(10 rats) and paste on non-meridian group(10 rats). METHODS: Osteoporosis model was prepared with injection of dexamethasonc. Antagonism osteoporosis acupoint paste(AOAP)on acupoints and non-neridian places was administrated to compare with oral recipe for kidney tonification in the treatment of osteoporosis. It took the changes in serum estrogen, bone density, calcitonin/parathyroid hormone(CT/PTH), thyrotropic hormone(TSH), freeT3andfreeT4asthemeasuringindexes. Immunocyto-chemistry stain and morphological measurement of TSH cell,follicular adenoma of thyroid gland(C cell) and thyroid globulin(Tg) were applied in pituitary gland and thyroid gland to observe the therapeutic results.MAIN OUTCOME MEASURES: ① Adjustment of the recipe for kidney tonification on thyroid function. ② Adjustment of the recipe for kidney tonification on bone density.RESULTS: AOAP improved the levels of estrogen, CT/PTH, TSH, free T3and free T4, increased bone density. The counts of TSH cell, C cell and Tg positive reaction substance were increased and they were colored deeply.CONCLUSION: AOPA, by "multiple relevant convergence reaction", adjusts and affects systematically the endocrinal function of bone metabolism,improves the levels of estrogen and CT/PTH, reverses the declining tendency of pituitary-thyroid axis function and improves thyroid function so as to weaken osteocyte in the bony absorption, enhance osteoblast in osteogenesis and increase bone density.

OBJECTIVESTo explore the effect of lead acetate on the apoptosis of rat brain neural cells and the relationship between the apoptosis and the bcl-2 as well as bax gene expression.

METHODSLead acetate was given to SD rats by intraperitoneal injection for 5 days at the dosage of 25, 50 and l00 mg/kg body weight respectively. The rates of apoptosis and the expression of bcl-2 (Bcl-2) and bax (Bax) in neural cells from cerebral cortex, hippocampus and carebellum were measured respectively by flow cytometry (FCM).

RESULTSThe rates of apoptosis in neural cells from cerebral cortex, hippocampus and cerebellum in every treatment group were significantly higher than that of control (P < 0.01), and there was a significant dose-response relationship (r = 0.998, 0.989 and 0.997 respectively). The expression of bcl-2 was significantly decreased, whereas bax was significantly increased, in neural cells from cerebral cortex, hippocampus and cerebellum in every lead acetate treatment group (FI) compared with the control group, and there was a significant dose-response relationship (r = -0.886, -0.787 and -0.832 respectively for bcl-2, r = 0.971, 0.988 and 0.991 respectively for bax). The value of Bcl-2/Bax in every treatment group decreased significantly compared with control, and there was a nice dose-response relationship (r = -0.863, -0.829 and -0.999, respectively). Correlation analysis showed that rates of apoptosis were inversely correlated with the expression of bcl-2 (r = -0.750, -0.509, and -0.667, respectively), whereas positively correlated with the expression of bax (r = 0.748, 0.56l, and 0.668, respectively). And there were inverse correlations between the rates of apoptosis and Bcl-2/Bax expression.

CONCLUSIONLead may induce apoptosis in rat brain neural cells through the down regulation of bcl-2 and the up regulation of bax gene expression.

Animals ; Apoptosis ; Brain ; cytology ; drug effects ; metabolism ; Female ; Male ; Organometallic Compounds ; administration & dosage ; pharmacology ; Proto-Oncogene Proteins ; biosynthesis ; Proto-Oncogene Proteins c-bcl-2 ; biosynthesis ; Rats ; Rats, Sprague-Dawley ; bcl-2-Associated X Protein

Objective To evaluate the skeletal stability of joint orthognathic and orthodontic treatment for cleft patients compared with non-cleft patients.Methods Fifteen cleft patient diagnosed with dental facial deformities underwent joint orthognathic and orthodontic treatment.Cephalometric analysis was carried out at T0 (before treatment),T1 (2 weeks after surgery),T2 (6 months after surgery) and T3 (24 months after surgery).The comparison of maxillary anterior-posterior and superiorinferior movement was performed with non-cleft orthognathic group (n =15) at the same follow-up time point.Results The maxilla was move forward for (3.8 ± 1.5) mm and downward for (2.4± 0.8) mm of cleft group.The relapse distance was (1.2±-0.7) mm (T1-T2:31.6％) in AP direction and (0.9±0.6) mm (T1-T2:37.5％) in vertical direction.At the time of 24 months after operation,the relapse distance was (1.0±0.5) mm (T1-T3:26.0％) in AP direction and (0.8±0.8) mm (T1-T3:33.3％) vertically.In non-cleft group,the maxilla was move (4.3±1.2) mm anteriorly and (2.2± 1.9) mm vertically.The relapse distance was (0.9±1.2) mm (T1-T2:20.9％) anteriorly and (0.8± 0.9) mm (T1-T2:36.6％) vertically at 6 months post-operatively.At 24 months after surgery the relapse distance was (1.1±0.6) mm (T1-T3:25.6％) anteriorly and (0.9±0.5) mm (T1-T3:40.9％) vertically.There were no statistical significant in both 6 months and 24 months follow-up between cleft and non-cleft group (P＞0.05).Conclusions There is no statistic difference of post-surgical relapse rate between cleft and non-cleft orthognathic and orthodontic treatments,although the relapse distances are greater than that in cleft group.

ObjectiveTo explore the protective effects of Dahuang Tangluo pills on early diabetic kidkdey disease （DKD） in db/db mice. MethodEight db/m mice were selected as the control group. Forty male db/db mice were selected and blood samples were collected via tail vein to measure fasting blood glucose （FBG）. Mice with FBG ≥ 16.7 mmol·L^-1， increased urine output， and persistent albuminuria were considered successful in model establishment. After successful modeling， they were randomly divided into a model group， a dapagliflozin group （1.5 mg·kg^-1·d^-1）， and high， medium， and low dose groups of Dahuang Tangluo pills （3.6， 1.8， 0.9 g·kg^-1·d^-1， respectively）， with eight mice in each group. All medication groups were administered orally， while the control and model groups were given an equal amount of distilled water by gavage daily. After continuous administration for 10 weeks， the survival status of the mice was observed， and their body weight， FBG， and kidney function-related indicators were measured. Inflammatory indicators in renal tissues were determined by enzyme-linked immunosorbent assay （ELISA）. Hematoxylin-eosin （HE） staining， Masson staining， and electron microscopy were used to observe the pathological changes in renal tissues in each group. Immunofluorescence was employed to examine the expression of advanced glycation end products （AGEs） and receptors for advanced glycation end products （RAGE） proteins. Real-time quantitative polymerase chain reaction （Real-time PCR） and Western blot were utilized to detect the gene and protein expression levels of AGEs， RAGE， inhibitor of nuclear factor-κB （NF-κB） kinase （IKK）， and NF-κB in the renal tissues of mice in each group. ResultCompared with control group， the model group showed a significant increase in body weight， FBG， serum creatinine （SCr）， urinary microalbumin/urine creatinine ratio （ACR）， total cholesterol （TC）， and triglycerides （TG） （P<0.05）. The levels of intercellular adhesion molecule-1 （ICAM-1）， interleukin-6 （IL-6）， and tumor necrosis factor-alpha （TNF-α） in renal tissues were significantly elevated （P<0.05）. Renal histopathological staining and electron microscopy revealed loose arrangement， gaps， structural disarray， mesangial proliferation， and significant fibrosis in renal tissues. Real-time PCR results showed a significant increase in the expression of RAGE， IKK， and NF-κB genes in renal tissues （P<0.05）. Immunofluorescence results demonstrated a significant increase in the expression of AGEs and RAGE proteins in renal tissues （P<0.05）. Western blot results showed a significant increase in the expression of AGEs， RAGE， IKK， and NF-κB proteins in renal tissues （P<0.05）. After drug intervention， compared with model group， the dapagliflozin group and the high-dose Dahuang Tangluo pills group showed significant reductions in body weight， FBG， SCr， and ACR （P<0.05）， and a significant decrease in TC in mouse serum （P<0.05）， while the high-dose Dahuang Tangluo pills group showed a significant decrease in TG in mouse serum （P<0.05）. All treatment groups showed a significant reduction in ICAM-1， IL-6， and TNF-α in renal tissues （P<0.05）. Renal histopathological staining and electron microscopy showed improved kidney injury， decreased collagen fiber deposition， and reduced mesangial proliferation in all treatment groups. Real-time PCR results showed a significant decrease in the expression of RAGE， IKK， and NF-κB genes in the dapagliflozin group and the high- and medium-dose Dahuang Tangluo pills groups （P<0.05）. Immunofluorescence results demonstrated a significant decrease in the expression of AGEs and RAGE proteins in the dapagliflozin group and the high- and medium-dose Dahuang Tangluo pills groups （P<0.05）. Western blot results showed a significant decrease in the expression of AGEs， RAGE， IKK， and NF-κB proteins in the dapagliflozin group and the high- and medium-dose Dahuang Tangluo pills groups （P<0.05）. ConclusionDahuang Tangluo pills can improve the pathological structure of the kidneys and reduce renal inflammation in DKD mice， possibly through inhibiting the AGEs/RAGE/IKK/NF-κB pathway.

Alzheimer's disease （AD） is a deleterious neurodegenerative disorder， which has become a significant public health concern and economic burden. The pathogenesis of AD is complex and involves several hypotheses such as amyloid β-protein （Aβ） deposition， Tau protein hyperphosphorylation， oxidative stress， and inflammation. There is an urgent need for a holism-based comprehensive intervention with multi-pathway， multi-level， and multi-target characteristics， which demonstrates the unique advantage of traditional Chinese medicine （TCM）. Therefore， it is of great significance to conduct and promote research on TCM treatment of AD. Danggui Shaoyaosan （DSS） from the Synopsis of Golden Chamber （《金匮要略》） by ZHANG Zhongjing （150 AD-219 AD） was originally designed for reliving gynecological ailments. It is a classic TCM formula that modulates liver and spleen and dispels blood stasis and water retention. Since the late 1980s when Japanese researchers reported its therapeutic effect on AD， it has been widely used in the clinic with clear effects. The elucidation of the mechanism of this formula helps exert its effects. Hereby， this paper reviewed relative research progress and made an analysis in terms of attenuating aberrant accumulation of Aβ and hyperphosphorylated Tau protein， anti-inflammatory and antioxidant activities， mediating neurotransmitters， ameliorating lipid metabolism， modulating gut microbiota， reduced neuron apoptosis， decreasing intracellular Ca²⁺ overloading， and increasing the expression of estradiol. This paper is expected to provide references for understanding the scientific connotation of DDS in the treatment of AD and lay a solid foundation for further investigation.

ObjectiveTo investigate the mechanism of Danggui Shaoyaosan （DSS） in the improvement of the cognitive ability of SAMP8 mice with Alzheimer's disease （AD） via regulating the ubiquitin-proteasome pathway （UPP）. MethodFifteen SAMR1 mice were used as a normal group， and 60 SAMP8 mice were randomly divided into a model group and DSS high， medium， and low-dose groups （57.6， 28.8， and 14.4 g·kg^-1·d^-1）， with 15 mice in each group. Intragastric administration was conducted for eight continuous weeks. Place navigation and spatial capacity were evaluated by Morris water maze. Pathological structure changes in neurons in the hippocampal CA1 area was detected by hematoxylin-eosin （HE） staining. The protein expression levels of hippocampal β-amyloid protein（Aβ） and phosphorylation（p）-Tau were determined by immunohistochemical staining （IHC） and enzyme-linked immunosorbent assay （ELISA）， and the mRNA and protein expression levels of hippocampal ubiquitin （Ub）， ubiquitin ligase E3 （E3）， 26S proteasome， ubiquitin carboxyl terminal hydrolase-1 （UCHL1）， and UCHL3 were determined by real-time fluorescent quantitative polymerase chain reaction （Real-time PCR） and Western blot， respectively. ResultAs compared with the normal group， the escape latency was prolonged in the model group （P<0.05） with the reduced number of crossing platform quadrants and time ratio in the platform quadrant （P<0.05）. The model group decreased neurons and condensed cell bodies in the CA1 area， and increased β-amyloid precursor protein （β-APP） and p-Tau positive cells （P<0.05）. In the model group， the protein expression levels of Aβ and p-Tau were increased （P<0.05）， the mRNA and protein expression levels of Ub were increased （P<0.05）， and the mRNA and protein expression levels of E3， 26S proteasome， UCHL1， and UCHL3 were decreased （P<0.05）. As compared with the model group， the escape latency was shortened in the DSS high and medium-dose groups （P<0.05） with an increased number of crossing platform quadrants and residence time ratio （P<0.05）. The pathological changes in CA1 of each DSS group were significantly improved， and the number of β-APP positive staining cells decreased （P<0.05）. The number of p-Tau positive staining cells decreased in the DDS medium and low-dose groups （P<0.05）. The protein expression levels of Aβ and p-Tau in each DDS group decreased （P<0.05）， and the mRNA expression level of Ub in each group decreased （P <0.05）. The mRNA expression levels of 26S， E3， and UCHL3 in the DDS high and medium-dose groups increased （P<0.05）， and the mRNA expression level of UCHL1 in the DDS medium-dose group increased （P<0.05）. The protein expression level of Ub in each DDS group decreased， and the protein expression levels of 26S， E3， UCHL1+3 in the DDS high and medium-dose groups increased （P<0.05）. ConclusionDSS can improve the cognitive ability of SAMP8 mice， and its mechanism may be related to the reduction of the abnormal deposition of Aβ and p-Tau via decreasing the expression of Ub and increasing that of E3， 26S， UCHL1， and UCHL3 in the UPP.

ObjectiveTo investigate the protective effect of Danggui Shaoyaosan （DSS）-contained serum on β-amyloid （Aβ）_1-40-injured rat adrenal pheochromocytoma PC12 cells and its mechanism in regulating ubiquitin-proteasome pathway （UPP）. MethodAβ_1-40 was used to intervene PC12 cells to prepare the cell models of Alzheimer's disease （AD）， and the experiment was divided into the blank， model， and DSS-contained serum high， medium， and low-dose groups （10%， 5%， and 2.5%）. Cell viability and apoptosis were detected using cell counting kit-8 （CCK-8） method and flow cytometry， respectively. The content of Aβ and p-Tau protein was determined by enzyme-linked immunosorbent assay （ELISA）. The ubiquitin （Ub）， ubiquitin ligase E3 （E3）， 26S proteasome， ubiquitin carboxyl terminal hydrolase1 （UCHL1）， and UCHL3 protein expressions of UPP were displayed using immunofluorescence cytochemistry （ICC）， and the mRNA and protein expression levels of Ub， E3-parkin， 26S， UCHL1， and UCHL3 were determined by real-time fluorescent quantitative polymerase chain reaction （Real-time PCR） and Western blot， respectively. ResultThe data of the CCK8 experiment verified that 5 μmol·L^-1 and 48 hours were the optimal conditions for modeling Aβ_1-40-injured PC12 cells. As compared with the blank group， the cell viability rate in the model group decreased （P<0.05） with an increased apoptosis rate （P<0.05）， the content of Aβ and p-Tau contents was elevated （P<0.05）， the mRNA and protein expression levels of Ub increased， and the mRNA and protein expression levels of 26S， E3， and UCHL1+3 decreased （P<0.05）. As compared with the model group， the cell viability rate in the DSS-contained medium-dose group increased （P<0.05）， whereas the apoptosis rate in each DSS-contained group decreased （P<0.05）. The content of Aβ in each DDS-contained group decreased （P<0.05）， and the content of p-Tau in the DDS-contained high and medium-dose groups decreased （P<0.05）. The mRNA expression level of Ub decreased， and that of 26S increased in each DDS-contained group （P<0.05）. The mRNA expression level of UCHL1 in the DDS-contained medium-dose group increased （P<0.05）， and the mRNA expression levels of E3 and UCHL 3 in the DDS-contained high and medium-dose groups increased （P<0.05）. The protein expression level of Ub in each DDS-contained group decreased， and the protein expression levels of 26S， E3， and UCHL1+3 in the DDS high and medium-dose groups increased. The DSS-contained serum medium-dose group exerted the optimal effect. ConclusionDSS-contained serum can increase cell viability rate， reduce cell apoptosis rate， eliminate Aβ and p-Tau protein deposits， and exert protective effects on Aβ_1-40-injured PC12 cells. Its mechanism may involve UPP via decreasing the expression of Ub and increasing that of 26S， E3， UCHL1， and UCHL3.

ObjectiveTo analyze the effects of Danggui Shaoyaosan （DSS） on the gut microbiota of the Alzheimer's disease （AD） model in SAMP8 mice based on 16S rDNA sequencing. MethodTwenty-four SAMP8 mice aged seven months were randomly divided into low-， medium-， and high-dose DSS groups （14.4， 28.8， 57.6 g·kg^-1·d^-1） and a model group according to a random number table， with six rats in each group. Six SAMR1 mice of the same age were assigned to the normal group. After intragastric administration for eight consecutive weeks， 16S rDNA sequencing was performed to detect the gut microbiota of feces in mice. Morris water maze was employed to assess the directional navigation and space exploration ability of mice. Nissl staining was performed to observe the pathological changes of neurons in the hippocampal CA1 area. Enzyme-linked immunosorbent assay （ELISA） was adopted to measure the protein content of hippocampal amyloid β-protein （Aβ） and hyperphosphorylated Tau （p-Tau）. ResultCompared with the normal group， the model group presented a declining α diversity （P<0.05）， markedly altered β diversity， prolonged escape latency （P<0.05）， reduced number of platform crossings and cumulative duration in the targeted quadrant （P<0.05）， decreased neurons and Nissl bodies in the CA1 hippocampal area， and up-regulated Aβ and p-Tau expression （P<0.05）. However， DSS intervention enhanced the α diversity， and medium- and high-dose DSS， especially the medium-dose DSS， could result in α diversity similar to the control group. Moreover， at the phylum level， the abundance of Firmicutes increased （P<0.05）， while the abundance of Bacteroidetes and Proteobacteria decreased （P<0.05）. At the genus level， the abundance of Lactobacillus and other genera increased （P<0.05）， while the abundance of Bacteroides， Helicobacterium， Rikenella， Parabacteroides， Sutterella， and Mucilaginibacter decreased （P<0.05）. The DSS groups also showed shortened escape latency （P<0.05）， increased number of platform crossings and cumulative duration in the targeted quadrant （P<0.05）， increased Nissl bodies （P<0.05）， and reduced Aβ and p-Tau content （P<0.05）. Pearson correlation analysis showed that the abundance of Mucilaginibacter， Bacteroides， and Sutterella was negatively correlated with the cognitive ability of SAMP8 mice， while the abundance of Lactobacillus and Butyricimonas was positively correlated with the cognitive ability of SAMP8 mice. ConclusionDSS can improve the cognitive ability of SAMP8 mice， and its mechanism may be related to the regulation of gut microbiota diversity and community composition.