Rapid determination of microbial contamination of Bupleurum chinense DC. decoction slices by ATP bioluminescence technology combined with statistical analysis methods / 药学学报

This study focuses on the microbial quality control of the Chinese herbal decoction pieces. In view of the shortcomings of traditional culture methods such as slow detection speed and inability to detect unculturable microorganisms, a new method based on ATP bioluminescence technology combined with statistical analysis methods was established to rapidly predict and quantitatively detect the total aerobic microbial count (TAMC) and total yeast and mold count (TYMC) contaminated Bupleurum chinense DC. decoction pieces. Based on the optimized ATP bioluminesence detection system, accurate detection of pure bacterial solution of Escherichia coli, Bacillus subtilis and Staphylococcus aureus can be achieved, with detection limits of 47.86, 89.13 and 1 862.09 CFU·mL-1, respectively. The detection time was 6.5 h, and the detection cost was as low as 2 yuan/time. The upper and lower warning limits of TAMC were determined by the misjudgment rates of 10% and 20%, respectively. And the warning limit of TYMC was determined by the misjudgment rate of 20%. The proposed crossing method could quickly predict the amount of microbial contamination in Bupleurum chinense DC. decoction pieces. The constructed partial least squares regression (PLSR) model could accurately quantify the quantity of microbial contamination in Bupleurum chinense DC. decoction pieces. The optimal PLSR prediction model for TAMC had a correction coefficient (R2) of 0.826, a root mean square error of correction set (RMSEE) of 0.468 and a root mean square error of cross-validation set (RMSECV) of 0.465. The R2, RMSEE and RMSECV in the prediction model of TYMC were 0.778, 0.543 and 0.541, respectively. The aim of this study is to establish a kind of rapid detection method and prediction models for the microbial limit of traditional Chinese medicine and Chinese herbal decoction pieces, and to provide a more convenient and sensitive detection technology for the microbial quality process control of traditional Chinese medicine products.

Upregulation of N-Type Voltage-Gated Calcium Channels Induces Neuropathic Pain in Experimental Autoimmune Neuritis.

Objective: Guillain-Barré syndrome (GBS) is a common autoimmune disease of the peripheral nervous system, and there is still no effective treatment for GBS. This investigation intends to figure out the effect and mechanism of N-type voltage-gated calcium (Cav2.2) channels on neuropathic pain in GBS. Methods: An experimental autoimmune neuritis (EAN) model was established in Lewis rats induced by myelin P253-78 peptide and complete Freund's adjuvant. Luxol fast blue (LFB) staining was used for observing the degree of cell infiltration and demyelination in the sciatic nerve of rats, ELISA for detecting IL-6 and TNF-α expression in the serum, qRT-PCR, and Western blot for measuring the expression of iNOS, MCP-1, and Cav2.2 in the sciatic nerve, respectively. Results: EAN led to significant decreases in the mechanical withdrawal threshold, thermal withdrawal threshold, and mechanical hyperalgesia threshold and an increase in the withdrawal threshold to cold stimulation. The serum IL-6 and TNF-α expression was significantly increased, and the mRNA and protein expression of iNOS, MCP-1, and Cav2.2 in the sciatic nerve were significantly increased in the EAN rats. However, silencing Cav2.2 expression could significantly reverse the above EAN-caused results. Conclusion: Silencing Cav2.2 expression can significantly reduce the clinical score, pathological injury, and mechanical allodynia, reducing the release of inflammatory factors, thus improving neuropathic pain in EAN rats.

Amphiphilic Functionalized Acupuncture Needle as SERS Sensor for In Situ Multiphase Detection.

Surface enhanced Raman spectroscopy (SERS) is a powerful spectroscopic technique with unique vibrational fingerprints, making it an ideal candidate for in situ multiphase detection. However, it is a great challenge to determine how to guide the SERS sensor to target molecules of interest in multiphase heterogeneous samples with minimal disturbance. Here, we present a portable ultrasensitive and highly repeatable SERS sensor for in situ multiphase detection. The sensor is composed of commercial Ag acupuncture needle and PVP-Au nanoparticles (Au NPs). The PVP on the Au NPs can adsorb and induce the Au NPs into a highly uniform array on the surface of the Ag needle because of its adhesiveness and steric nature. The Au NPs-Ag Needle system (Au-AgN) holds a huge SERS effect, which is enabled by the multiple plasmonic couplings from particle-film and interparticle. The PVP, as the amphiphilic polymer, promotes the target molecules to adsorb on surface of the Au-AgN whether in the oil phase or in the water phase. In this work, the Au-AgN sensor was directly inserted into the multiphase system with the laser in situ detection, and SERS detection at different spots of the Au-AgN sensor provided Raman signal of targets molecule in the different phase. In situ multiphase detection can minimize the disturbance of sampling and provide more accurate information. The facile fabrication and amphiphilic functionalization make Au-AgN sensor as generalized SERS detection platform for on-site testing of aqueous samples, organic samples, even the multiphase heterogeneous samples.

[Clinical research on fire filiform needle combined with mild moxibustion for postherpetic neuralgia].

OBJECTIVE: To compare efficacy differences between fire filiform needle combined with mild moxibustion and gabapentin combined with sham acupuncture for postherpetic neuralgia (PHN). METHODS: One hundred cases of PHN were randomly divided into a needle group and a medicine group, 50 cases in each one. In the needle group, pricking method of fire filiform needle was given at the Ashi points, and then mild moxibustion was applied for 15 min. In the medicine group, the oral administration of gabapentin capsule and sham acupuncture at non-acupoints in the distal end of lesions were applied. The treatment was required for 21 days in both groups. The visual analogue score (VAS) was recorded before treatment and on the 1st day, 2nd day, 3rd day, 6th day, 9th day and 12th day of treatment. The most severity of pain within last 24 h, preset severity of pain, immediate analgesia effect and starting time of pain relief were observed, also the efficacy was assessed and improvement of symptoms was observed in the follow-up visit. RESULTS: The total effective rate was 94.0% (47/50) in the fire filiform needle group, which was superior to 86.0% (43/50) in the medicine group (P < 0.05). Compared with medicine group, the VAS of the most severity of pain within last 24 h was obviously reduced after the 2nd treatment in the fire filiform needle group while that of present severity of pain was relieved after the 1st treatment (both P < 0.05). The immediate analgesia effect in the fire filiform needle group was obviously superior to that in the medicine group in the first three times of treatment (all P < 0.05). The average time of pain relief was (3.91 +/- 0.82) days in the fire filiform needle group, which was significantly earlier to (6.53 +/- 1.13) days in the medicine group (P < 0.05). 26 cases were cured in the fire filiform needle group in the follow-up visit, which was superior to 2 cases in the medicine group (P < 0.05). The improvement of VAS, pain range and sleep quality in the needle group were also superior to those in the medicine group (all P < 0.05). The direct medical cost in the fire filiform needle group was (232.32 +/- 48.108) yuan, which was significantly lower than (466.00 +/- 41.09) yuan in the medicine group (P < 0.05). There was only one case of adverse effect in the medicine group during the treatment. CONCLUSION: The fire filiform needle combined with mild moxibustion could obviously relieve the pain in PHN patients, which has superior immediate analgesia effect and pain relieving time compared with gabapentin, which also has less adverse effects and cheap cost.

Hybridaphniphyllines A and B, Daphniphyllum alkaloid and iridoid hybrids suggestive of Diels-Alder cycloaddition in Daphniphyllum longeracemosum.

Two Daphniphyllum alkaloid and iridoid hybrids with the hitherto unknown decacyclic fused skeletons, hybridaphniphyllines A and B, along with one diamino Daphniphyllum alkaloid, daphnicyclidin I, were isolated from dried stems and leaves of Daphniphyllum longeracemosum. Their structures were elucidated on the basis of extensive spectroscopic analysis, and the absolute configuration of daphnicyclidin I was deduced by the CD exciton chirality method. A biogenetic pathway for 1 and 2 involving natural Diels-Alder cycloaddition is proposed.

[Effects of Chinese herbal medicine Guanxinkang on expression of PPARγ-LXRα-ABCA1 pathway in ApoE-knockout mice with atherosclerosis].

OBJECTIVE: To observe the effects of Guanxinkang (GXK) decoction, a compound traditional Chinese herbal medicine, on expressions of peroxisome proliferator-activated receptor γ (PPARγ), liver X receptor α (LXRα) and ATP-binding cassette transporter A1 (ABCA1) in apolipoprotein E (ApoE)-knockout mice with atherosclerosis. METHODS: Fourteen 6-week-old C57BL/6 J mice were used as normal control group. Seventy 6-week-old ApoE-knockout mice receiving a high-cholesterol diet to induce atherosclerosis were randomly divided into untreated group, simvastatin group and low-dose (concentration of crude drugs at 0.864 g/mL), medium-dose (crude drugs at 1.728 g/mL) and high-dose (crude drugs at 3.456 g/mL) GXK groups. After treated with the drugs for eight weeks continuously, the livers and aortas of mice were separated. The expressions of PPARγ, LXRα and ABCA1 were measured by real-time quantitative polymerase chain reaction and Western blotting respectively. RESULTS: Compared with the normal control group, mRNAs and proteins of PPARγ, LXRα and ABCA1 over-expressed in the untreated group (P<0.05). After the treatment, GXK decoction and simvastatin decreased the expressions of PPARγ, LXRα and ABCA1 (P<0.05). High-dose GXK decoction had more marked effects than low- and medium-dose GXK and simvastatin. CONCLUSION: The PPARγ-LXRα-ABCA1 pathway is involved in lipid regulation and inflammation activities. Over-expression of the genes has complicated effects on atherosclerosis in ApoE-knockout mice with high-cholesterol diet. GXK decoction has anti-inflammatory and anti-matrix metalloproteinase activities by regulating PPARγ, LXRα and ABCA1 interactions in the ApoE-knockout mice.

[Experience of Shi's Bian stone comprehensive therapy for treating periarthritis of shoulder].

The experiences of Doctor SHI An-li, who created the SHI's Bian stone comprehensive therapy, for treating periarthritis of shoulder are introduced. There are three important points: (1) emphasizing selection of points and manipulation; (2) using the Bian stone comprehensive therapy flexibly; (3) practicing the elongated needle on the obstinate periarthritis of shoulder, and the manipulation process are explained in detail.

[Effects of Chinese herbal medicine Guanxinkang on lipid metabolism and serum C-reactive protein, amyloid A protein and fibrinogen in apolipoprotein E-knockout mice with atherosclerosis].

OBJECTIVE: To observe the effects of Guanxinkang (GXK) decoction, a compound traditional Chinese herbal medicine, on serum lipids and apolipoprotein A I (ApoA I), apolipoprotein B (ApoB), apolipoprotein E (ApoE), C-reactive protein (CRP), serum amyloid A protein (SAA) and fibrinogen (Fbg) concentrations of ApoE-knockout mice with atherosclerosis, and to explore the mechanism of GXK decoction in anti-atherosclerosis. METHODS: Seventy 6-week-old ApoE-knockout mice receiving a high-cholesterol diet were used to induce atherosclerosis and were randomly divided into 5 groups: untreated group, simvastatin group and low- (drug concentration is 0.864 g/mL), medium- (1.728 g/mL), and high-dose (3.456 g/mL) GXK groups. Another fourteen 6-week-old C57BL/6J mice were used as the normal control. Two 12-week-old mice were randomly selected from the normal control and the ApoE-knockout mice respectively to observe vulnerable plaque in the mouse's aortic by hematoxylin-eosin staining. Blood was collected from venous plexus of eye socket after gavage of corresponding drugs once daily for 8 weeks continuously, and then the serum was separated. Triglyceride (TAG) and total cholesterol (TC) were measured by enzyme-coupled assay; low-density lipoprotein cholesterol (LDL-C) and high-density lipoprotein cholesterol (HDL-C) were measured by selective precipitation method. Serum levels of ApoA I and ApoB were determined by turbidimetry. Double-antibody sandwich enzyme-linked immunosorbent assay was used to detect ApoE, CRP, SAA and Fbg concentrations in serum. RESULTS: Compared with the normal control group, the levels of serum TC, TAG, LDL-C, ApoB, CRP, SAA and Fbg in the untreated group were increased (P<0.05), and the serum concentrations of HDL-C, ApoA I and ApoE in the untreated group were decreased (P<0.05). After treatment, GXK decoction and simvastatin improved the dyslipidemia by increasing the concentrations of ApoA I and HDL-C and decreasing the concentrations of TC, TAG, LDL-C, ApoB, CRP, SAA and Fbg (P<0.05). The high-dose GXK decoction had the most marked effects on SAA and Fbg and the serum lipids compared with the low-dose and medium-dose GXK and simvastatin. CONCLUSION: GXK decoction may not only provide an active effect on hyperlipidemia, but also down-regulate the levels of serum CRP, SAA and Fbg. GXK decoction exerts an anti-atherosclerosis effect in ApoE-knockout mice.

[Literature quality evaluation criteria for Clinical Practice Guideline of Evidence-based Acupuncture and Moxibustion].

In Clinical Practice Guideline of Evidence-based Acupuncture and Moxibustion, following principles and methods of evidence-based medicine, in combination with characteristics of acupuncture and moxibustion science, primary literature quality assessment criteria and corresponding scale were stipulated and were repeatedly seeked advice from experts and proved, finally, forming the assessment criteria: (1) Evaluation criteria of literature quality for RCT; (2) Evaluation criteria of literature quality for non-randomly controlled trials; (3) Evaluation criteria of literature quality for cases-study trials.

[Trial for replacing fire needle by filiform needle].

OBJECTIVE: To promote a new kind of alternative needle of filiform-fire needle with fire needle. METHODS: The common three-headed fire needle, thick fire needle, less-thick fire needle, fine fire needle, single-filiform needle and three-filiform-needle were used by 20 medical staffs, and the time of heating needles was recorded. The advantages of using filiform needles compared with common fire needles were evaluated. RESULTS: The time of heating single and three-filiform needles was shorter than the other four common fire needles (all P < 0.01). CONCLUSION: Filiform needle is better than the common fire needle in terms of production process,materials, and heating time, etc. Therefore, filiform needles can replace common fire needles for pricking.

[Clinical observation on strong stimulation of acupoints around eyes for treatment of adolescent myopia].

OBJECTIVE: To observe the therapeutic effect and influential factors of strong stimulation of acupoints around eyes by reducing method of twisting the needle for treatment of adolescent myopia, and to compare the different curative effects of both weak and strong stimulus intensities and wearing glasses in daytime. METHODS: According to the principle of knowing and permission, the patients were divided into a strong stimulation group (60 cases, 120 eyes), a weak stimulation group (60 cases, 120 eyes) and a wearing glasses group (60 cases, 120 eyes). Both the strong stimulation group and the weak stimulation group were treated by reducing method of twisting the needle, with Cuanzhu (BL 2), Yuyao (EX-HN 4), Sizhukong (TE 23), Sibai (ST 2) and Taiyang (EX-HN 5) selected. Each acupoint was twisted 360 and 90 times per min for the strong stimulation group, and 90 degrees and 60 times per min for the weak stimulation group; the wearing glasses group were not treated with acupuncture, but with wearing glasses in daytime. RESULTS: After treatment with the reducing method of twisting the needle, the vision improved (P < 0.01), with more obviously increased in the strong stimulation group than the weak stimulation group (P < 0.01), and there was no significant change of vision in the wearing glasses group. The age and the myopic degree of the patient before treatment were closely related with the change of vision after treatment (P < 0.01). CONCLUSION: The reducing method of twisting the needle has a definite clinical therapeutic effect on adolescent myopia; strong stimulation has a better effect than the weak one; the age and the myopic degree of the patient possibly influence the therapeutic effect.

[Evaluation of literature quality of acupuncture for treatment of herpes zoster and approach to the laws of treatment].

OBJECTIVE: To assess the quality of literature of clinical studies on acupuncture in treatment of herpes zoster. METHODS: The literatures between 1994-2006 were searched by means of electronic retrieval. Type and methodology, general condition, diagnosis of diseases and enrolled and excluded criteria, assessment of sample content, treatment condition, criteria for assessment of therapeutic effects, following-up, etc. in clinical studies are evaluated according to principles and methods of clinical epidemiology and evidence-based medicine. RESULTS: Of the 399 literatures enrolled, only 8 were authentic randomized controlled trials (RCTs), 20 quasi-randomized controlled trials, 66 non-randomized concurrent controlled trials and 277 narrative studies, 70 had clear diagnostic criteria, 16 mentioned enrolled or excluded criteria, 287 had clear criteria for therapeutic effects, 107 reported follow-up, 2 had the description of health economical index, 9 reported adverse reaction. CONCLUSION: At present, correct randomization, concealment, blinding and placebo-control, and the RCTs with generally accepted criteria for assessment of diagnosis and therapeutic effects, safety evaluation and rational design of follow-up are needed. It is indicated by preliminary study of the literatures that blood-letting puncture and cupping at Ashi points are main methods for treatment of herpes zoster.

[Analysis on the current situation of establishing control group in clinical studies of acupuncture and moxibustion for treatment of periarthritis of shoulder].

OBJECTIVE: To probe into the current situation and problems of establishing control group in clinical studies of acupuncture and moxibustion for treatment of periarthritis of shoulder at home. METHODS: To retrieve the literatures of establishing control groups in clinical studies of acupuncture and moxibustion for treatment of periarthritis of shoulder in magazines at home, and based on key problems in establishment of control group, make a Excel form to take out relative contents in the literatures and classify, summarize and statistically analyze. RESULTS: In establishing control group in clinical studies of acupuncture and moxibustion for treatment of periarthritis of shoulder, a lot of problems exist, most studies do not have the basis of establishing control group, and also do not establish control group for the purpose of the study, making the whole test have error in the stage of program design, and leading to unreliable for the results of clinical control studies. CONCLUSION: In clinical researches of acupuncture and moxibustion, a standard for guiding establishment of control group is eagerly needed.