Effect of Bushen Chushi() decoction combined with platelet-rich plasma on regulating TGF-β1 and Smad-1 expression in serum of knee osteoarthritis patient / 中国骨伤

OBJECTIVE@#To investigate the effect of Bushen Chushi decoction combined with platelet-rich plasma(PRP) to treat knee osteoarthritis(KOA) in early and middle stage and its regulation on TGF-β1 and Smad-1 expression in serum.@*METHODS@#Total of 45 patients with KOA in early and middle stage from May 2020 to April 2022 were treated and divided into control group and observation group. In control group, there were 30 patients including 12 males and 18 females, aged from 43 to 69 years old with an average of(57.3±6.5) years old and disease duration ranged from 1.5 to 5.0 years with an average of(3.8±1.7) years, and there were 8 cases in gradeⅠ, 13 cases in gradeⅡ, and 9 cases in grade Ⅲ according to Kellgren-Lawrence Grade, PRP 5 ml was injected into knee joint on the first day of No1, 3 week together for 2 times. In the observation group, there were 15 cases including 7 males and 8 females, aged from 45 to 70 years old with an average of (56.7±6.2) years old and disease duration ranged from 1.8 to 5.7 years with an average of (4.0±1.8) years, there were 4 cases in gradeⅠ, 9 cases in gradeⅡand 4 cases in grade Ⅲ according to the Kellgren-Lawrence Grade, PRP 5 ml were injected into knee joints that the time and frequency were the same as those in the control group, and at the same time Bushen Chushi decoction orally were taken 1 dose per day with a total of 28 doses. All patients were treated for four weeks. Visual analogue scale(VAS) and Lequesne MG score before and after treatment were used to evaluate improvement of knee pain and joint function. The TGF-β1 and Smad-1 levels in serum were measured before and after treatment in two groups. The incidence of complications in two groups was observed.@*RESULTS@#All patients were followed up for 26 to 30 days with an average of (28.0±0.6) days. There was no significant difference in VAS and knee Lequesne MG scores between two groups before treatment(P>0.05). The scores of VAS and knee Lequesne MG on the first day after treatment in both groups were lower than those before treatment(P<0.05). The VAS and knee Lequesne MG scores in observation group were lower than those in control group(P<0.05) on the first day after treatment. The TGF-β1 level in serum after treatment were higher significantly than that before treatment in two groups(P<0.05). After treatment, TGF-β1 level in serum in observation group were lower than those in control group with statistically significant differences(P<0.05). The Smad-1 levels in serum after treatment in observation group were higher significantly than that in control group(P<0.05). The levels of Smad-1 were not statistically significant between before and after treatment(P>0.05). There was no significant difference in postopertaive complications between two groups (P>0.05).@*CONCLUSION@#The efficacy of Bushen Chushi decoction combined with PRP in treatment of early and middle KOA is better than that of PRP injection alone. The combined treatment could reduce TGF-β1 level and increase Smad-1 level in serum, which may be a mechanism to inhibit inflammation and alleviate cartilage degeneration to some extent.

Response to roxadustat in a patient undergoing long-term dialysis and allergic to erythropoiesis-stimulating agents: A case report.

BACKGROUND: Hypoxia-inducible factor prolyl hydroxylase inhibitor is a new class of drugs for treating renal anemia. It is a second-generation hypoxia-inducible factor prolyl hydroxylase-2 (PHD2) inhibitor. Roxadustat can effectively increase hemoglobin in patients with dialysis-dependent chronic kidney disease, with an adverse events profile comparable to that of epoetin alfa. We administered roxadustat to a maintenance hemodialysis patient who was allergic to erythropoiesis-stimulating agents (ESAs) and depended on blood transfusion for five years. After applying Roxadustat, the patient's anemia improved significantly. CASE SUMMARY: A 77-year-old Chinese man had type 2 diabetes for 16 years, underwent maintenance hemodialysis for five years, and had fatigue for five years. Laboratory tests showed severe anemia (hemoglobin concentration of 42 g/L). The patient was administered a subcutaneous injection of ESAs before dialysis. He suffered an allergic shock immediately and fainted. His blood pressure dropped to undetectable levels. He was not administered ESAs henceforth. The patient was prescribed iron supplements and received blood transfusions occasionally for five years. His hemoglobin concentration ranged from 42-68 g/L. After taking six weeks of oral roxadustat three times weekly (100 mg TIW), the patient's hemoglobin concentration increased significantly, and his symptoms decreased. We adjusted the doses of roxadustat, and the hemoglobin concentration was maintained between 97 and 126 g/L. CONCLUSION: Oral roxadustat is effective in treating anemia in maintenance hemodialysis patients who cannot be administered ESAs.

The effects of electro-acupuncture on learning, memory and stem cell differentiation in the hippocampus of rats modeling chronic cerebral ischemia / 中华物理医学与康复杂志

Objective:To observe the effect of electro-acupuncture (EA) on learning, memory and the differentiation of neural stem cells in the hippocampus in the presence of chronic cerebral ischemia.Methods:A total of 120 male Sprague-Dawley rats had chronic cerebral ischemia induced by bilateral ligation of the common carotid arteries. The model was successfully established in 104 of them, and they were randomly divided into a model group and an EA group, each of 52. The EA group was given 20 minutes of EA at acpoints Baihui and Dahui every day for 7 days, followed by an interval of 2 days. The current output was 1mA and the frequency was 15Hz. The model group was not given any intervention. One, 2, 4 and 6 weeks after the modelling, 6 rats from each group were injected with BrdU, and any proliferation and differentiation of neural stem cells was observed. The rats′ learning and memory were also evaluated using the Morris water maze, and neurogenesis of in the dentate gyrus was observed using BrdU+ NeuN and BrdU+ GFAP double-labelled immunofluorescence.Results:The learning and memory of the EA group were significantly better than those of the model group 2, 4 and 6 weeks after the modelling. After two weeks BrdU+ NeuN and BrdU+ GFAP-positive cells were found in the granule cell layer of the hippocampus, and compared with the model group, there were siginificantly more such neurons in the EA group. The gliocyte levels were not significantly different.Conclusion:Electro-acupuncture can improve learning and memory in the face of chronic cerebral ischemia by promoting the differentiation of neural stem cells.

Comprehensive analysis of Polygoni Multiflori Radix of different geographical origins using ultra-high-performance liquid chromatography fingerprints and multivariate chemometric methods.

Polygoni Multiflori Radix (PMR) is increasingly being used not just as a traditional herbal medicine but also as a popular functional food. In this study, multivariate chemometric methods and mass spectrometry were combined to analyze the ultra-high-performance liquid chromatograph (UPLC) fingerprints of PMR from six different geographical origins. A chemometric strategy based on multivariate curve resolution-alternating least squares (MCR-ALS) and three classification methods is proposed to analyze the UPLC fingerprints obtained. Common chromatographic problems, including the background contribution, baseline contribution, and peak overlap, were handled by the established MCR-ALS model. A total of 22 components were resolved. Moreover, relative species concentrations were obtained from the MCR-ALS model, which was used for multivariate classification analysis. Principal component analysis (PCA) and Ward's method have been applied to classify 72 PMR samples from six different geographical regions. The PCA score plot showed that the PMR samples fell into four clusters, which related to the geographical location and climate of the source areas. The results were then corroborated by Ward's method. In addition, according to the variance-weighted distance between cluster centers obtained from Ward's method, five components were identified as the most significant variables (chemical markers) for cluster discrimination. A counter-propagation artificial neural network has been applied to confirm and predict the effects of chemical markers on different samples. Finally, the five chemical markers were identified by UPLC-quadrupole time-of-flight mass spectrometer. Components 3, 12, 16, 18, and 19 were identified as 2,3,5,4'-tetrahydroxy-stilbene-2-O-ß-d-glucoside, emodin-8-O-ß-d-glucopyranoside, emodin-8-O-(6'-O-acetyl)-ß-d-glucopyranoside, emodin, and physcion, respectively. In conclusion, the proposed method can be applied for the comprehensive analysis of natural samples.

Therapeutic observation of cranial suture acupuncture in treating depression / 针灸推拿医学（英文版）

Objective:To observe the clinical efficacy and safety of cranial suture acupuncture plus paroxetine in treating depression,and to discuss the action mechanism of this acupuncture method.Methods:One hundred depression patients were allocated to an observation group and a control group according to the random number table,with 50 cases in each group.The control group was intervened by oral administration of paroxetine tablets,20 mg each time,once a day for successive 6 weeks;the observation group was additionally given cranial suture acupuncture,once a day for 6 weeks.They were scored by Hamilton depression scale-17 (HAMD-17) before the treatment and respectively after 1-week,2-week,4-week and 6-week treatment.The clinical efficacy and safety were also observed.Results:After 6-week treatment,the total effective rate was 94.0％ in the observation group versus 78.0％ in the control group,and the between-group difference was statistically significant (P＜0.01).The HAMD-17 scores respectively after 1-week,2-week,4-week and 6-week treatment were significantly lower than the score before the treatment in the observation group (all P＜0.05);the HAMD-17 scores respectively after 2-week,4-week and 6-week treatment were significantly different from the score before the treatment in the control group (all P＜0.05).There were significant differences in the HAMD-17 score between the two groups respectively after 4-week and 6-week treatment (both P＜0.05).Conclusion:Cranial suture acupuncture plus paroxetine can ease the symptoms of depression,with faster onset and more significant therapeutic efficacy compared with paroxetine alone.

Gastrodin Reduces the Severity of Status Epilepticus in the Rat Pilocarpine Model of Temporal Lobe Epilepsy by Inhibiting Nav1.6 Sodium Currents.

Temporal lobe epilepsy (TLE) is one of the most refractory types of adult epilepsy, and treatment options remain unsatisfactory. Gastrodin (GAS), a phenolic glucoside used in Chinese herbal medicine and derived from Gastrodia elata Blume, has been shown to have remarkable anticonvulsant effects on various models of epilepsy in vivo. However, the mechanisms of GAS as an anticonvulsant drug remain to be established. By utilizing a combination of behavioral surveys, immunofluorescence and electrophysiological recordings, the present study characterized the anticonvulsant effect of GAS in a pilocarpine-induced status epilepticus (SE) rat model of TLE and explored the underlying cellular mechanisms. We found that GAS pretreatment effectively reduced the severity of SE in the acute phase of TLE. Moreover, GAS protected medial entorhinal cortex (mEC) layer III neurons from neuronal death and terminated the SE-induced bursting discharge of mEC layer II neurons from SE-experienced rats. Furthermore, the current study revealed that GAS prevented the pilocarpine-induced enhancement of Nav1.6 currents (persistent (INaP) and resurgent (INaR) currents), which were reported to play a critical role in the generation of bursting spikes. Consistent with this result, GAS treatment reversed the expression of Nav1.6 protein in SE-experienced EC neurons. These results suggest that the inhibition of Nav1.6 sodium currents may be the underlying mechanism of GAS's anticonvulsant properties.

Experience of Professor LIU Zhe for Treating Essential Tremor with Acupuncture / 浙江中医药大学学报

[Objective]To study the clinical experience of Professor LIU Zhe treatment of essential tremor with acupuncture.[Methods]The paper states Professor LIU's understanding of treating essential tremor from the pathogenesis, acupuncture points and acupuncture manipulations, then summarizes the principle, regularity, and the features of acupuncture manipulations for Professor LIU treating essential tremor of different types of syndrome, so as to elaborate the clinical experience of Professor LIU. [Result] Professor LIU Zhe considers the age-dependent deficiency of qi and blood and insufficient liver and kidney syndrome to be a common one of essential tremor, as an experienced doctor, he distinguishes different types of syndrome from sign, tongue, pulse and so on. He places great emphasis on nourishing the bone marrow and tonifying liver and kidney. According to the symptoms select acupuncture points with the effects of nourishing the bone marrow and calming the liver to stop the wind, tonifying liver and kidney and supplementing qi and nourishing blood. Professor Liu sets a high value on selecting points of head and makes use of the point of Fengchi with unique acupuncture manipulations to relax cerebrovascular. In clinically, the curative effects are remarkable. [Conclusion]The unique method for treating essential tremor of Professor LIU gives some inspirations for clinic. It is worth of a deep study and spreading.

Anticonvulsant effect of Rhynchophylline involved in the inhibition of persistent sodium current and NMDA receptor current in the pilocarpine rat model of temporal lobe epilepsy.

Rhynchophylline (RIN) is a significant active component isolated from the Chinese herbal medicine Uncaria rhynchophylla. Several studies have demonstrated that RIN has a significant anticonvulsant effect in many types of epilepsy models in vivo. However, the mechanisms of the anticonvulsant effect remain elusive. Using combined methods of behavioral testing, immunofluorescence and electrophysiological recordings, we characterized the anticonvulsant effect of RIN in a pilocarpine-induced status epilepticus (SE) rat model of temporal lobe epilepsy (TLE) and investigated the underlying cellular mechanisms. In one set of experiments, rats received RIN treatment prior to pilocarpine injection. In a second set of experiments, rats received RIN treatment following the onset of stage 3 seizures. Pretreatment and posttreatment with RIN effectively reduced the seizure severity in the acute phase of TLE. Furthermore, RIN protected medial entorhinal cortex (mEC) layer III neurons from neuronal death and terminated spontaneous epileptiform discharge of mEC layer II neurons in SE-experienced rats. Whole-cell voltage-clamp recordings indicated that RIN inhibited neuronal hyperexcitability via inhibition of the persistent sodium current (INaP) and NMDA receptor current. Immunofluorescence experiments also demonstrated that RIN rectified the pilocarpine-induced upregulation of Nav1.6 and NR2B protein expression. In conclusion, our results identified RIN as an anticonvulsant agent that inhibited ictal discharge via INap and NMDA receptor current inhibition.

One-pot ß-cyclodextrin-assisted extraction of active ingredients from Xue-Zhi-Ning basing its encapsulated ability.

Xue-Zhi-Ning (XZN) is a traditional Chinese medicine formula, containing active ingredients with poor solubility in water, which has been demonstrated to be helpful for patients with hyperlipidemia. One-pot ß-cyclodextrin (ß-CD)-assisted extraction of active ingredients from XZN has been carried out to develop an efficient and eco-friendly extraction process. Five active compounds--rubrofusarin gentiobioside, 2,3,5,4'-tetrahydroxy-stilbene-2-O-ß-D-glucoside, emodin, nuciferine and quercetin--were identified by UPLC/DAD/MS and used as indexes to evaluate the process optimized by an orthogonal test. The results showed that addition of ß-CD significantly enhanced the extraction ratios of all five components. The enhancement of extraction ratios was positively correlated with the apparent formation constants between ß-CD and the compounds. The study also showed that the stabilities and dissolution rates of the active ingredients were improved in the presence of ß-CD. This one-pot ß-cyclodextrin-assisted extraction has the potential to be applied in pharmaceutical preparations directly.

Polyanthumin, a novel cyclobutane chalcone trimmer from Memecylon polyanthum.

A novel unusual trimmer chalcone, polyanthumin (1), together with five known compounds myricetin 3-O-(3″-O-galloyl)-α-l-rhamnopyranoside (2), sulfuretin (3), fustin (4), gallic acid (5), and ethyl gallate (6), was isolated from the dry stems of Memecylon polyanthum H.L. Li. Among them, compound 1 is a new chalcone trimmer with a novel cyclobutane skeleton in nature. Compounds 3 and 4 are flavonoids carrying a single 7-OH in A ring, which provided the first example of these class flavonoids from the family Melastomataceae. In addition, the antitumor activities for 2-4 were reported for the first time in this study. The antitumor effects of the isolated compounds 1-6 in vitro were assayed by the SRB method using human cancer K562 cells, with the inhibition rates ranging from 39.4% to 54.5% at 100 µg/ml. The IC50 values of compounds 1 and 3 for the inhibition of K562 cell proliferation were determined to be 45.4 and 30.5 µg/ml, respectively. To the best of our knowledge, compound 1 was the second sample as chalcone trimer. In addition, the antitumor activities for 2-4 were reported for the first time in this study.

The effects of compound danshen dripping pills and human umbilical cord blood mononuclear cell transplant after acute myocardial infarction.

OBJECTIVES: The low frequency of survival of stem cells implanted in the myocardium after acute myocardial infarction may be caused by inflammation and oxidative stress in the myocardial microenvironment. We evaluated the effects of a traditional Chinese medicine, Compound Danshen Dripping Pills, on the cardiac microenvironment and cardiac function when used alone or in combination with human umbilical cord blood mononuclear cell transplant after acute myocardial infarction. MATERIALS AND METHODS: After surgically induced acute myocardial infarction, rabbits were treated with Compound Danshen Dripping Pills alone or in combination with human umbilical cord blood mononuclear cell transplant. Evaluation included histology, measurement of left ventricular ejection fraction and fractional shortening, leukocyte count, count of green fluorescent protein positive cells, superoxide dismutase activity, and malondialdehyde content. RESULTS: Combination treatment with Compound Danshen Dripping Pills and human umbilical cord blood mononuclear cell transplant significantly increased the survival of implanted cells, inhibited cardiac cell apoptosis, decreased oxidative stress, decreased the inflammatory response, and improved cardiac function. Rabbits treated with either Compound Danshen Dripping Pills or human umbilical cord blood mononuclear cells alone had improvement in these effects compared with untreated control rabbits. CONCLUSIONS: Combination therapy with Compound Danshen Dripping Pills and human umbilical cord blood mononuclear cells may improve cardiac function and morphology after acute myocardial infarction.

A UPLC-MS/MS method for in vivo and in vitro pharmacokinetic studies of psoralenoside, isopsoralenoside, psoralen and isopsoralen from Psoralea corylifolia extract.

ETHNOPHARMACOLOGICAL RELEVANCE: The dried fruit of Psoralea corylifolia L. has been used to prevent and treat vitiligo, osteoporosis, arthralgia and asthma in Traditional Chinese Medicine for some 1600 years. Psoralen (P), isopsoralen (IP), psoralenoside (PO) and isopsoralenoside (IPO) are the major coumarins and coumarin-related benzofuran glycosides in Psoraleae Fructus, which have been reported to show estrogen-like activity, osteoblastic proliferation accelerating activity, antitumor effects and antibacterial activity. The first aim of this study is to develop a rapid, sensitive and selective ultra performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) approach for simultaneous determination of PO, IPO, P and IP in rat plasma and samples collected from in vitro incubation experiments. The second aim is to investigate the pharmacokinetic properties of PO, IPO, P and IP after oral administration of Psoralea corylifolia extract (PCE) to rats. The third aim is to confirm the biotransformation of PO to P or IPO to IP under gastrointestinal conditions. MATERIALS AND METHODS: A UPLC-MS/MS method with a C18 column and a mobile phase of methanol-0.1% aqueous formic acid was validated according to the criteria in FDA guidelines about bioanalytical method, which was developed to investigate the pharmacokinetic behavior of PO, IPO, P and IP from PCE and the metabolic pathways of PO to P or IPO to IP. RESULTS: The criteria for establishment of a new UPLC-MS/MS method including selectivity, linearity, accuracy, precision, extraction recovery, matrix effect and stability were validated. This method was successfully applied to the quantitative determination of PO, IPO, P and IP in biological samples collected from both in vitro incubations and in vivo rat experiments. After oral administration of PCE to rat, pharmacokinetic parameters of these four compounds indicated that in vivo biotransformation may occur between PO and P or IPO and IP. Purified benzofuran glycosides fraction (PBGF), containing only PO and IPO, was orally administered to rats to further confirm the biotransformation of PO to P or IPO to IP under gastrointestinal conditions. An in vitro incubation study elucidated that PO and IPO were metabolized to P and IP by intestinal microflora through de-glucosylation. CONCLUSIONS: This paper developed a rapid, sensitive and selective UPLC-MS/MS method for simultaneous determination of PO, IPO, P and IP from PCE in biological samples, and investigated on their comprehensive in vivo and in vitro pharmacokinetic studies. These obtained results showed that the metabolism by intestinal bacteria plays an important role in pharmacological effects of orally administered PCE.

[Simultaneous determination of fumonisins B1 and B2 in traditional Chinese medicines by high-performance liquid chromatography-tandem mass spectrometry].

A rapid and sensitive analytical method was developed for the simultaneous determination of fumonisins B1 and B2 in traditional Chinese medicines by HPLC-MS/MS. The detection limits for fumonisins B1 and B2 were 0.25 ng x mL(-1) corresponding to 2 microg x kg(-1) in samples. Recoveries from different samples spiked with fumonisins B1 and B2 at levels ranging from 0.2 to 3 mg x kg(-1) were 84.0%-96.1% and 86.3%-99.3%, respectively. Among the total of 34 samples purchased from local markets, ten samples of which were visibly moldy samples due to inappropriate storage, and 24 were normal samples. The results showed that 6 of the visibly moldy samples and 5 of the normal samples were contaminated with total fumonisins at levels ranging 82.4-2349 microg x kg(-1) and 102-729 microg x kg(-1), respectively.

Kinetics and mechanism of 2,3,5,4'-tetrahydroxystilbene-2-O-ß-d-glycoside (THSG) degradation in aqueous solutions.

The hydrolytic kinetics and degradation mechanism of 2,3,5,4'-tetrahydroxystilbene-2-O-ß-d-glycoside (THSG) extracted from Radix Polygoni Multiflori (a commonly used official Chinese herbal Heshouwu), were investigated using reversed-phase high-performance liquid chromatography (HPLC) and liquid chromatography-mass spectrometry (LC-MS). The influences of pH (1.5-9.9), temperature (25-60°C) and irradiation on the hydrolysis of THSG were studied in aqueous solutions. The results showed that the degradation of THSG was pH-, temperature- and irradiation-dependent and all followed first-order kinetics. The effect of temperature on the rate of THSG degradation was characterized using the Arrhenius equation. Maximum stability of THSG was found at pH 1.5 (t(0.5)=47.57 d). THSG was unstable in alkaline and irradiation conditions. The active energy (E(a)) of THSG degradation in aqueous solution at pH 6.8 (most frequently adopted extract solvent) under lucifugal and irradiation conditions was 47.7kJmol(-1) and 25.3kJmol(-1), respectively. Three hydrolytic products of THSG were identified by LC-MS. Cis-trans isomerism took place under irradiation, and hydrolysis took place in acid-base conditions. Moreover, further oxidation on aglycon occurred after hydrolytic cleavage of phenolic glycoside in acidic conditions. The possible hydrolytic pathways of THSG are proposed.