Effect of different therapies of Chinese medicine on the expressions of c-Fos and c-Jun proteins in hippocampus of rats with post-stroke depression

BACKGROUND ： c-fos and c-jun, the important immediate early genes （IEG）, are regarded as the markers for the location and function of neuronal activity, as well as the third signal messengers, they couple the stress stimulation and the gene expression in neuron, and hippocampus is involved in the process of signal transmission after stress stimulation induced depression. OBJECTIVE： To observe the therapeutic effects of Bushen Yiqi （tonifying kidney to benefit qi）, Huoxue Huayu （promoting blood circulation to dissipate blood stasis） and Ditan Kaiqiao （eliminating phlegm for resuscitation） on the expressions of c-Fos and c-Jun proteins in hippocampus and spontaneous behaviors of rats with post-stroke depression （PSD）, and compare the results with those of fluoxetine, which is known to have definite effect on depression. DESIGN： A randomized controlled tna SETTING ： Zhejiang College of Traditional Chinese Medicine MATERIALS ： The trial was completed in Zhejiang College of Traditional Chinese Medicine from January to July in 2003. Fifty-six healthy adult Wistar male rats of clean grade, weighing （250±50） g, were randomly divided into 7 groups with 8 rats in each group： control group, model group, forced swimming group, Bushen Yiqi group; Huoxue Huayu, Ditan Kaiqiao group and fluoxetine group. The Bushen Yiqi Tang contained Renshen, Huangqi, Heshouwu, Gouqi, Shudi, etc., crude drugs 1 800 g/L. The Huoxue Huayu Tang contained Danshen, Chuanxiong, Chishao, Yujin, etc., crude drugs 3 600 g/L. The Ditan Kaiqiao Tang contained Banxia, Danxing, Changpu, Yuanzhi, etc., crude drug 1 000 g/b METHODS： ① Except the control group and forced swimming group, rats in the other groups were made into PSD models by deligating the bilateral common carotid artedes permanently. ② Rats in the control group, model group and forced swimming group were intragastncally perfused by saline （3 mL for each time）; those in the Bushen Yiqi group, Huoxue Huayu, Ditan Kaiqiao group and fluoxetine group were intragastncally perfused with Bushen Yiqi Tang （18 g/kg）, Huoxue Huayu Tang （9 g/kg）, Ditan Kaiqiao Tang （9 g/kg） and fluoxetine （2.5 mg/kg） respectively, once a day. ③ At 55 days after model establishment, rats in the forced swimming group were managed according to the Porsolt＇s method. They were placed in water for 15 minutes, and then taken out and dned, no moving-time within 5 minutes was recorded at drying and 24 hours after drying. ④ Measurement of spontaneous behaviors： Except the forced swimming group, the spontaneous behaviors and activities （including horizontal and vertical movements） of rats were observed with the Open-Field method at 28, 42 and 56 days after administration in the other groups. ⑤ The expressions of c-Fos and coJun proteins in hippocampus were determined with the immunohistochemical method, the relative sectional area ratio and average objective gray value of c-Fos and c-Jun positive cells in hip- pocampus were measured with the computerized image analytical system. MAIN OUTCOME MEASURES： The spontaneous behaviors of rats, the relative sectional area ratio and average objective gray value of c-Fos and c-Jun positive cells in hippocampus were observed. RESULTS： Of the 56 rats, 1 died in the forced swimming group, and finally 55 rats were involved in the analysis of results. ① Results of spontaneous activities： At 28 days, the times of crossing movements were obviously fewer in the model group and fluoxetine group [（69.00±37.01）, （98.11 ±36.68） times/3 minutes] than in the control group [（128.44±16.85） times/3 minutes, P 〈 0.01, 0.05], but those in the Bushen Yiqi group, Huoxue Quyu group and Ditan Kaiqiao group had no obvious differences as compared with those in the control group （P 〉 0.05）. At 42 and 56 days, the times of crossing movements were obviously more in the Bushen Yiqi group, Huoxue Quyu group and Ditan Kaiqiao group [（106.44±31.24）, （117.20±23.95）, （134.80±28.18）, （136.36±40.95） times/3 minutes; （117.33±35.91）, （129.60 ±23.78）, （131.90 ±26.81）, （136.09±28.34） times/3 minutes] than in the model group [（64.00±17.51）, （72.86±20.68） times/3 minutes, P 〈 0.01]. The times of rearing movements had no obvious differences among the groups for the three times （P 〉 0.05）. ② The no moving-time within 5 minutes 24 hours after drying was obviously longer than that at drying in the forced swimming group. ③ The average objective gray values of c-Fos positive cells were not obviously different in the Bushen Yiqi group and Ditan Kaiqiao group from the control group （P 〉 0.05）, but lower in the model group than in the control group （69.84±9.82, 75.78±5.89, P 〈 0.01）, and higher in the forced swimming group than in the control group （85.97±10.99, P 〈 0.01）; all higher in the fluoxetine group, Bushen Yiqi group, Huoxue Quyu group and Ditan Kaiqiao group than in the model group （81.27±10.73, 74.04±8.34, 83.29±9.89, 70.14±4.92, P 〈 0.05-0.01）. The average objective gray values of c-Jun positive cells were obviously lower in the Bushen Yiqi group than in the control group （68.11 ±6.89, 79.58±5.86, P 〈 0.01）, but all higher in the other groups than in the control group （84.68±7.15, 81.34 ±8.36, 97.51±10.55, 85.68±9.25, 86.19±10.98, P 〈 0.05-0.01）; Those were obviously higher in the fluoxetine group, Huoxue Quyu group and Ditan Kaiqiao group than in the model group （P 〈 0.05-0.01 ）, lower in the Bushen Yiqi group than in the model group （P 〈 0.05）, all obviously lower in the Bushen Yiqi group, Huoxue Quyu group and Ditan Kaiqiao group than in the fluoxetine group （P 〈 0.01）. The relative sectional area ratios of c-Fos and c-Jun positive cells had no obvious differences among the groups （P 〉 0.05）. CONCLUSION ： The methods of Bushen Yiqi, Huoxue Quyu and Ditan Kaiqiao can effectively treat PSD in rats, and the results were equivalent with those of fluoxetine, the actions of the above-mentioned drugs may correlated with their regulation to c-Fos and c-Jun expressions in hippocampus. PSD animal models can be successfully established by both permanent deligation of bilateral common carotid arteries and forced swimming, and the models induced by the former has similar basic cerebrovascular lesions as human stroke in clinic.

CLINICAL OBSERVATION ON TREATMENT OF STUBBORN HICCUP BY ACUPUNCTURE WITH DIFFERENT NEEDLE RETAINING TIME

Purpose:Observe the effect of different duration of needle-retention on acupuncture treatment of stubborn hiccup. Methods: 85 cases of stubborn hiccup patients are randomly divided into control group (n=40) and treatment group(n=45) according to their visiting time. For patients of control group, acupuncture needles (in both scalp points and body acupoints) are retained for 30 min, while for patients of treatment group, the needles inserted in the body acupoints are retained for 1 hr, and that inserted in scalp-point is retained for more than 6 hrs. The treatment is conducted once daily, continuously for 5 times. Results: The cure rate of treatment group is 88.9% (that for deficiency syndrome is 85.3%), and that of control group is 65% (that for deficiency syndrome is 53.3%).There is a significant difference between two groups in the cure rate (P<0.05). The cure rates for excess syndrome of two groups both are 100%.Conclusion: Longer retention of acupuncture needle has a better therapeutic effect for deficiency-syndrome type hiccup.

Efficacy assessment for different acupuncture therapies in the treatment of frozen shoulder

Objective To explore better therapies for the treatment of frozen shoulder. Methods One hundred and seventy-four cases were divided into a filiform needle group （56 cases）, an electroacupuncture group （57 cases） and a warming needle group （61 cases） according to the randomized, controlled and single-blind study principles. Jiānqián （肩前 Extra）, Jiānyú （肩髃 LI 15）, Jiānliáo （肩髎 TE 14）, Nàoshū （臑俞 SI 10）, Wàiguān （外关 TE 5）, Hégǔ （合谷 LI 4） were used in all three groups but treated with filiform needle, electroacupuncture and warming needle technique respectively. The needles were retained for 30 min. It was given once every other day and 5 times constituted as one course. Pain indices and activity degree of shoulders were measured and recorded before and after treatment every time. Results The total effective rate was 93.0% （53/57） in the electroacupuncture group and 95.1% （58/61） in the warming needle group, both superior to that of 78.6% （44/56） in the filiform needle group （both P〈0.01）, but there was no significant difference between electroacupuncture group and warming needle group （P〉0.05）. After one course of treatment, the decline indices of shoulder pain of electroacupuncture group （4.28±0.22） and warming needle group （3.74±0.17） were both significantly greater than that of filiform needle group （2.78±0.18）（both P〈0.01）. And the decline indices of electroacupuncture group was also greater than that of warming needle group （P〈0.05）. The improvements of shoulder activity degree of warming needle group （76.92±5.53） and electroacupuncture group （60.37±3.80） were both greater than that of filiform needle group （42.50±3.67） （both P〈0.01）. And shoulder activity degree of warming needle group was also greater than that of electroacupuncture group （P〈0.01）. After one course of treatment, improvement of shoulder activity degree and decline indices of shoulder pain in these three groups were better than that after the first time treatment （all P〈0.01）. Conclusion All these three acupuncture therapies can achieve good therapeutic effects for frozen shoulder. The therapeutic effects of electroacupuncture and warming needle groups are superior to that of filiform needle group. All these three therapies could significantly reduce patients’ pain and improve their shoulder activity degree. The analgesic effect of electroacupuncture is the best, and the shoulder activity degree improved by warming needle is the best. The improvement of clinical therapeutic effect mainly depends on the therapy and the treatment times when the same acupoints are selected and the condition of illness are similar.

Knee osteoarthritis treated with acupuncture based on syndrome differentiation:a randomized controlled trial

Objective To compare the efficacy differences between acupuncture and western medicine in treating osteoarthritis of knee joints.Methods One hundred and twenty cases were randomly divided into an acupuncture group and a western medication group with 60 cases in each group.In the acupuncture group,acupuncture was applied mainly at Nèixīyǎn（内膝眼 EX-LE 4）,Dúbí（犊鼻 ST 35）,Yīnlíngquán（阴陵泉 SP 9） and Yánglíngquán（阳陵泉 GB 34） plus other acupoints selected according to syndrome differentiation.In the medication group,Glucosamine Hydrochloride capsules were orally given 3 times daily with 2 capsules each time.Western Ontario and McMaster Universities Osteoarthritis Index（WOMAC）,Medical Outcomes Study 36-Item Short Form Health Survey（SF-36,Chinese version） were used to assess therapeutic effects before treatment,after 4 weeks treatment and 9 weeks after treatment,respectively.Results After 4 weeks of treatment,the total effective rates were 86.7%（52/60） and 88.3%（53/60） in the acupuncture group and the western medication group,respectively,without significant difference（P〈0.05）.In 9 weeks after treatment,the total effective rates were 83.3%（50/60） and 61.7%（37/60） in the acupuncture group and the western medication group,respectively,with significant difference between the two groups（P〈0.05）.According to WOMAC and SF-36,the scores were all improved in both groups（P〈0.05,P〈0.01）,with significant difference in follow-up visit of 9 weeks after treatment between the two groups（P〈0.05,P〈0.01）,but not after 4 weeks of treatments（both P〈0.05）.Conclusion Short-term of therapeutic effects of acupuncture and oral administration of Glucosamine Hydrochloride on osteoarthritis of knee joints are similar,but long-term efficacy in the acupuncture group is better than that in the western medication group.

Is Manipulative Therapy Clinically Necessary for Relief of Neck Pain？A Systematic Review and Meta-Analysis

Objective：To summarize and critically assess the efficacy of Eastern and Western manipulative therapies for the treatment of neck pain in adults. Methods：A search of Pub Med/MEDLINE, the Cochrane Central Register of Controlled Trials, Clinical Trials.gov, EMBASE, etc. from their inception date to January 2014 with Chinese, Japanese, and Korean databases. Two reviewers independently selected randomized controlled trials（RCTs） with negative control or blank control, extracted data and assessed methodological quality. Meta-analysis and levels of evidence were performed by Revman5.1 and Grades of Recommendations Assessment, Development and Evaluation（GRADE） approach. Results：Nineteen clinical trials with adequate randomization were included in this review, 11 of them had a low risk of bias. The primary outcome for shortterm pain had no significant differences, however, the secondary outcome, only the Numerical Pain Rating Scale（NPRS） score of intermediate-term [n=916, pooled mean differences（MD） =–0.29, P=0.02], the Neck Disability Index（NDI） score of short-term（n=1,145, pooled MD=–2.10, P〈0.01）, and intermediate-term（n=987, pooled MD=–1.45, P=0.01） were significantly reduced with moderate quality evidence. However, it supported the minimally clinically important difference（MCID） of the Visual Analogue Scale and NPRS pain score to be 13 mm, while NDI was 3.5 points. The meta-analysis only suggested a trend in favor of manipulative therapy rather than clinical significance. Conclusions：The results do not support the existing evidences for the clinical value of Eastern or Western manipulative therapy for neck pain of short-term follow-up according to MCIDs. The limitations of our review related to blinding, allocation concealment and small sample size.