[Electroacupuncture relieved neuropathic pain by suppressing L-Arg/NO/cGMP pathway in the lumbar spinal cord in rats with spared nerve injury].

OBJECTIVE: To observe the effect of electroacupuncture (EA) on changes of expression of L-Arg transporter 2 (CAT-2) mRNA and nitric oxide synthase (iNOS) mRNA and protein and contents of NO and cGMP of L4-L6 segments of spinal cord in rats with spared nerve injury (SNI), so as to reveal its mechanism underlying reducing neuropathic pain. METHODS: A total of 120 male SD rats were randomly divided into sham operation, model, EA and NOS inhibitor (N omega-Nitro-L-arginine methyl ester hydrochloride, L-NAME) groups, with 30 rats in each group. The neuropathic pain model was established by ligating and cutting the tibial nerve and the common peroneal nerve. EA (2 Hz, 1-3 mA) was applied to "Weizhong" (BL40) and "Huantiao" (GB30)on the damaged hindlimb for 30 min, once daily from day 11 to 17 after SNI. Rats of the L-NAME group received i.p. of L-NAME (60 mg·kg-1·d-1) for 7 consecutive days. The mechanical pain threshold (PT) was determined before and 10 and 16 d after SNI, respectively. The expression le-vels of CAT-2 mRNA and iNOS mRNA, and iNOS protein in the L4-L6 segments of the spinal cord were detected by using reverse transcription - polymerase chain reaction (RT-PCR) and Western blot, respectively, and the contents of NO and cGMP of L4-L6 assayed using nitrate/nitrite reductase method and radioimmunoassay, respectively. RESULTS: After modeling, the PT was significantly decreased on day 10 and 16 after SNI in comparison with the sham operation group and their own baseline data of pre-operation in each group (P<0.01), and remarkably increased in the EA and L-NAME groups relevant to the model group on day 16 (P<0.01, P<0.05). Compared with the sham operation group, the expression levels of CAT-2 mRNA and iNOS mRNA and protein, as well as the contents of NO2－/NO3－and cGMP were signi-ficantly up-regulated in the model group (P<0.05, P<0.01). Following EA intervention, the levels of CAT-2 mRNA and iNOS mRNA and iNOS protein, and NO2－/NO3－and cGMP contents were all reversed in both EA and L-NAME groups (P<0.05, P<0.01). The effect of EA was significantly superior to that of L-NAME in raising the PT on day 16 after SNI (P<0.05), but obviously inferior to that of L-NAME in down-regulating the expression of CAT-2 mRNA and iNOS mRNA and protein (P<0.05). No significant differences were found between the EA and L-NAME groups in down-regulating NO2－/NO3－ andcGMP contents (P>0.05). CONCLUSION: EA intervention can effectively relieve neuropathic pain in SNI rats, which may be closely related to its function in suppressing L-Arg/NO/cGMP pathway in the lumbar spinal cord.

A New Compound Heterozygous Mutation Of BSCL2 In A Chinese Zhuang Ethnic Family With Congenital Generalized Lipodystrophy.

PURPOSE: This study aims to report the clinical features of an infant with CGL in a Chinese Zhuang ethnic family, whose family members were discovered to carry new pathogenic mutations in the BSCL2. PATIENTS AND METHODS: In this study, we report clinical and molecular investigations of CGL disease in a family of 4 members (parents and two sons). We used whole exome sequencing (WES) in the family to examine the genetic cause of the disease. RESULTS: The proband presented with skin pigmentation, hypertriglyceridemia and diabetes. WES identified a previously unreported compound heterozygous mutation in the BSCL2 (c.545_546insCCG heterozygous mutation and exon 3 heterozygous deletion) in the proband. His mother is a heterozygous carrier of the c.545_546insCCG mutation and his father and brother are carriers of the exon 3 heterozygous deletion. CONCLUSION: Compound heterozygous mutation of the BSCL2 (new c.545_546insCCG heterozygous mutation and new exon 3 heterozygous deletion) was detected in the proband with characteristic clinical manifestations of CGL2.

[Effect of Electroacupuncture Intervention on CaMK II-CREB Pathway in Spinal Cord in Rats with Spared Nerve Injury].

OBJECTIVE: To observe the effect of electroacupuncture (EA) stimulation of "Weizhong" (BL 40)-"Huantiao" (GB 30) on expression of phosphorylated calcium/calmodulin dependent protein kinase II (p-CaMK II) and cAMP response element binding protein (p-CREB) in the spinal cord in rats with spared nerve injury (SNI), so as to explore its mechanism underlying easing neuropathic pain. METHODS: Sixty SD rats were randomly divided into five groups: control (sham-operation) , model, EA, AP-5 (a NMDA receptor antagonist) and L-NAME (a non-selective nitric oxide synthase, NOS inhibitor) (n = 12 in each group). The neuropathic pain model was established by sectioning the right tibal nerve and common peroneal nerve. EA intervention (2 Hz, 1 mA, increasing 1 mA/10 min) was applied to "Weizhong" (BL 40) and "Huantiao" (GB 30) on the injured side for 30 min, once a day for 7 days. Rats of the AP-5 and L-NAME groups were treated by intragastric administration of AP-5 (0.7 mg · kg(-1) · d(-1)) and L-NAME (60 mg · kg(-1) · d(-1)) respectively from the 11 th day after operation, once daily for 7 days. The mechanical pain thresholds were measured before the SNI procedure (baseline) and at the 10th and 16th day after the procedure. The expression of p-CaMK II protein and p-CREB protein and gene of the spinal cord (L4-L6 segments) was determined by Western blot and fluorescence quantitative-polymerase chain reaction (PCR), separately. RESULTS: In comparison to the control group, the mechanical pain threshold was significantly decreased in the model group (P < 0.01). After EA intervention, the mechanical pain thresholds of the EA, AP-5 and L-NAME groups were obviously increased (P < 0.01, P < 0.05) on day 16 post SNI procedure. The expression levels of p-CaMK II and p-CREB proteins and CREB mRNA in the spinal cord were significantly higher in the model group than in the control group (P < 0.05). Compared with the model group, the expression levels of spinal p-CaMK II and p-CREB proteins and CREB mRNA were obviously down-regulated in the EA group (P < 0.05), but not in the AP-5 group and the L-NAME group (P > 0.055. CONCLUSION: EA intervention of BL 40-GB 30 may alleviate pain in neuropathic pain rats, which may be related to its effects in down-regulating spinal CaMK II-CREB pathway function.

[Effect of electroacupuncture intervention on N-methyl-D-aspartic acid receptor expression in spinal cord in rats with chronic constrictive injury of the sciatic nerve].

UNLABELLED: OBJECTIVE; To observe the changes of N-methyl-D-aspartic acid (NMDA) receptor expression of spinal cord after electroacupuncture (EA) intervention in rats with chronic constrictive injury (CCI) of the sciatic nerve so as to reveal the mechanism underlying improvement of neuropathic pain. METHODS: Sixty male SD rats were randomly divided into sham operation (sham) group, CCI model group and EA group (n = 20). CCI model was established by ligature of the right sciatic nerve with a piece of surgical chromic suture. For rats of the sham group, the right sciatic nerve was just isolated without ligature. The rats of the EA group were given with EA stimulation of "Weizhong" (BL 40) and "Huantiao" (GB 30) on the injured side at a frequency of 2 Hz, electric current of 1-3 mA for 30 min (increasing 1 mA every 10 min). The treatment was conducted once a day from the 11th day to the 20th day after modeling. NMDA receptor 2 B subunit (NR 2 B) protein and mRNA expression levels in the spinal cord were determined by immunohistochemistry, Western blot and reverse transcription (RT)-polymerase chain reaction (PCR), respectively, and spinal NMDA receptor subunit 1 (NR 1) protein and mRNA expression levels were measured by Western blot and RT-PCR, respectively. RESULTS: In comparison with the sham group, NR 1 protein and mRNA expression levels of the model group in the spinal cord were considerably upregulated after CCI (P < 0.01, P < 0.05). In comparison with the model group NR 1 protein and mRNA expression levels of the EA group in the spinal cord were evidently down-regulated (P < 0.05). No significant changes of NR 2 B protein and mRNA expression after CCI and EA stimulation were found after immunohistochemistry, Western blot and RT-PCR measurements (P > 0.05). CONCLUSION: EA intervention is effective in alleviating neuropathic pain in CCI rats, which may be closely related to its effects in lowering functional activity of NR 1 protein and mRNA in the spinal cord.

[Influence of acupuncture on the motor ability in training-induced fatigue mice].

OBJECTIVE: To observe the influence of acupuncture on the motor ability of training-induced fatigue mice so as to explore its mechanism underlying acupuncture-induced improvement of physical training fatigue. METHODS: Sixty Kunming mice were divided into normal control, training model and acupuncture groups, with 20 mice in each. The training-fatigue model was established by forcing the mouse to swim in a water tank for 30 min/d in the 1st week, 60 min/d in the 2nd week, 90 min/d in the 3rd week and 120 min/d in the 4th and 5th weeks. "Guanyuan" (CV 4), and bilateral "Zusanli" (ST 36) and "Shenshu" (BL 23) were punctured with filiform needles, and with the needle twirled for about 30 s and retained for 5 min. Thetreatment was conducted once daily for 2 weeks. The duration of exhaustion training was recorded. Serum lactic acid (LA), lactate dehydrogenase (LDH) and creatine kinase (CK) contents were detected by LA oxidase method, immune-suppressive assay and enzyme-linked immunoassay, respectively. RESULTS: Compared with the training-fatigue (model) group, the duration of the exhausted swimming of the acupuncture group was obviously longer (P < 0.05). In comparison with the control group, serum LA, LDH and CK contents were increased considerably in the model and acupuncture groups (P < 0.05, P < 0.01), while compared with the model group, serum LDH level of acupuncture group was increased significantly (P < 0.05), and serum CK level of acupuncture group decreased obviously (P < 0.05). No significant difference was found between the model and acupuncture groups in serum LA content (P > 0.05). CONCLUSION: Acupuncture can effectively improve the motor ability of the training-fatigue mice, which may be closely relevant to its effects in upregulating LDH activity and reducing serum CK.