Electroacupuncture and A-317491 depress the transmission of pain on primary afferent mediated by the P2X3 receptor in rats with chronic neuropathic pain states.

P2X is a family of ligand-gated ion channels that act through adenosine ATP. The P2X3 receptor plays a key role in the transmission of neuropathic pain at peripheral and spinal sites. Electroacupuncture (EA) has been used to treat neuropathic pain effectively. To determine the role of EA in neuropathic pain mediated through the P2X3 receptor in dorsal root ganglion neurons and the spinal cord, a chronic constriction injury (CCI) model was used. Sprague-Dawley rats were divided into four groups: sham CCI, CCI, CCI plus contralateral EA, and CCI plus ipsilateral EA. The mechanical withdrawal threshold (MWT) and thermal withdrawal latency (TWL) were recorded. Furthermore, the expression of the P2X3 receptor was evaluated through Western blotting and immunofluorescence. The effects of EA and A-317491 were investigated through the whole-cell patch-clamp method and intrathecal administration. Our results show that the MWT and TWL of EA groups were higher than those in the CCI group, whereas the expression of the P2X3 receptor was lower than that in the CCI group. However, no significant difference was detected between the two EA groups. EA depressed the currents created by ATP and the upregulation of the P2X3 receptor in CCI rats. Additionally, EA was more potent in reducing mechanical allodynia and thermal hyperalgesia when combined with A-317491 through intrathecal administration. These results show that both contralateral and ipsilateral EA might inhibit the primary afferent transmission of neuropathic pain induced through the P2X3 receptor. In addition, EA and A-317491 might have an additive effect in inhibiting the transmission of pain mediated by the P2X3 receptor.

[Effect of "Buqi Yixue" needling on neurological function and nerve conduction velocity in patients with diabetic peripheral neuropathy].

OBJECTIVE: To observe the efficacy of "Buqi Yixue "needling on clinical symptoms, neurological function and nerve conduction velocity in patients with diabetic peripheral neuropathy. METHODS: Eighty-six patients with diabetic peripheral neuropathy numbness and pain were equally randomized into control group and treatment group. The patients of the control group received basic treatment and oral administration of Cilostazol (50 mg/time, 2 times/d) and Epalrestat (50 mg/time, 3 times/d). The patients of the treatment group received acupuncture stimulation of Danzhong (CV17), Qihai (CV6), Pishu (BL20), Quchi (LI11), etc., for 30 min, once every day, on the basic treatment. The treatment was conducted for 8 successive weeks. The scores of Traditional Chinese Medicine (TCM) symptoms, Toronto clinical scoring system (TCSS) and nerve conduction velocity (NCV) were detected before and after the treatment. RESULTS: After the treatment, the scores of TCM symptoms and TCSS were considerably decreased (P<0.05), and the NCV was significantly increased (P<0.05) in both groups compared with those of their own pre-treatment. The the-rapeutic effect of the treatment group was significantly superior to that of the control group in lowering the scores of TCM symptoms and TCSS, and in up-regulating the NCV (P<0.01). CONCLUSION: "Buqi Yixue" needling is effective in improving clinical symptoms and increasing NCV in patients with diabetic peripheral neuropathy.

Oligonucleotide chip, real-time PCR and sequencing for genotyping of hepatitis B virus.

AIM: To compare the oligonucleotide chip, real-time PCR and sequencing for genotyping of hepatitis B virus in Chinese patients with chronic hepatitis B. METHODS: Mixture of samples with different genotypes and clinical serum samples from 126 chronic hepatitis B patients was tested for hepatitis B virus genotypes by oligonucleotide chip, real-time PCR and sequencing of PCR products, respectively. Clinical performances, time required and costs of the three assays were evaluated. RESULTS: Oligonucleotide chips and real-time PCR detected 1% and 0.1% genotypes, respectively, in mixed samples. Of the 126 clinical samples from patients with chronic hepatitis B, genotype B was detected in 41 (33%), 41 (33%) and 45 (36%) samples, and genotype C in 76 (60%), 76 (60%) and 81 (64%) samples, by oligonucleotide chip, real-time PCR and sequencing, respectively. Oligonucleotide chip and real-time PCR detected mixed genotypes B and C in 9 samples. Real-time PCR was the rapidest and cheapest among the three assays. CONCLUSION: Oligonucleotide chip and real-time PCR are able to detect mixed genotypes, while sequencing only detects the dominant genotype in clinical samples.

Detection of the rtA181V/T and rtN236T mutations associated with resistance to adefovir dipivoxil using a ligase detection reaction assay.

BACKGROUND: Adefovir dipivoxil (ADV) is effective for treatment of chronic hepatitis B virus (HBV) infection, but long-time ADV therapy leads to drug resistance because of HBV reverse transcriptase mutations. We developed a sensitive and specific method for detecting the rtA181V/T and rtN236T mutations associated with ADV resistance in chronic hepatitis B patients, based on a ligase detection reaction (LDR). METHODS: HBV templates were amplified by polymerase chain reaction (PCR), followed by LDR and electrophoresis on a sequencer. The assay was evaluated using 165 serum samples, and plasmid controls. RESULTS: In a mixture of wild-type and mutant plasmids, the assay could detect mutant plasmid at 1%. Complete concordance between the PCR-LDR assay and sequencing analysis was observed for 141 of 148 samples (95.3%) at codon 181, and 143 of 148 samples (96.6%) at codon 236. Discordant results were confirmed to be consistent with the PCR-LDR assay by subclone sequencing. Seventeen samples could not be detected by both of the methods due to low HBV DNA levels. CONCLUSIONS: The PCR-LDR assay can sensitively and specifically detect the rtA181V/T and rtN236T mutations, and may be used for monitoring ADV resistance in patients infected with HBV.

[Clinical significance of AFP-L3 variants determined by micro centrifugal column].

OBJECTIVE: To evaluate the clinical significance of AFP-L3 in patients with hepatocellular carcinoma. METHODS: Serum AFP-L3 variants were separated by micro centrifugal column, and detected by chemiluminescence. RESULTS: AFP and AFP-L3 levels were higher in patients with hepatocellular carcinoma than those in patients with chronic hepatitis (P<0.001); as a diagnostic target, the sensitivity and specificity of AFP-L3 were 72.3 percent and 97.2 percent, respectively. Eight patients with hepatitis have higher AFP-L3, but none of them were found with carcinoma by CT three months later. CONCLUSION: AFP-L3 is very useful in the diagnosis of patients with hepatocellular carcinoma.