Molecular mechanism of Yaotu Granules in the prevention and treatment of lumbar disc degeneration based on RNA-seq technology / 中国组织工程研究

BACKGROUND:The Herbal Compound formula Yaotu Granules for lumbar disc degeneration has achieved satisfactory efficacy, but the underlying mechanism remains unclear. OBJECTIVE: To explore the possible molecular mechanisms ofYaotu Granules for lumbar disc degeneration based on transcriptome sequencing (RNA-seq). METHODS: Ten New Zealand white rabbits were randomly divided into control and experimental groups (n=5 per group), followed by intragastric injection of 20 mL of normal saline and 20 mL of water decoction of Yaotu Granules, respectively, twice daily for consecutive 1 week. The different drug serums were prepared using serum samples obtained from the common carotid artery at 2 hours after the last injection. Human nucleus pulposus cells were cultured in DMEM containing 10% different drug serums to observe the cellular morphology in each group. The cell viability was detected by trypan blue staining. The passage 3 human nucleus pulposus cells were pretreated for 48 hours, the mRNA level was detected using RNA-seq, and the differentially expressed mRNA was screened by RNA-seq. Subsequently, the differential gene expression, Gene Ontology (GO) enrichment analysis and Pathway enrichment analysis were performed. RESULTS AND CONCLUSION: In the experimental group, there were round or fusiform cells with abundant cytoplasm and clear nuclei that showed smooth surface and complete nuclear membrane; elongated rough endoplasmic reticulum arranged regularly, and less mitochondrion, scattered lysosomes and filaments were visible in the cytoplasm. In the control group, spindle and polygonal cells were found, and the large nuclei with 2 or 3 nucleoli and slightly rough surface were observed; there were mildly dilated endoplasmic reticulum, and few mitochondria with incomplete membrane. The adherent rate, generation time and cell viability of passage 1, 2 and 3 cells in the experimental group were significantly better than those in the control group (P < 0.05). Sequencing results found that there were 464 differentially expressed genes including 143 upregulated, and 321 downregulated genes. GO analysis revealed that the differentiated genes were mainly concentrated on cell regulation and metabolism. Pathway analysis found that mainly differentiated genes focused on the metabolism related pathways. These findings suggest that the differentially expressed gene profile of Yaotu Granules for lumbar disc degeneration is obtained by RNA-seq technology.Yaotu Granules mainly upregulate extracellular matrix metabolism-related genes and downregulate polysaccharide synthesis related genes in the prevention and treatment of lumbar disc degeneration, which provides basis for further research.

Preparation of intervertebral disc degeneration models by percutaneous acupuncture rotary-cut under different anesthesia methods / 中国组织工程研究

BACKGROUND:Different anesthesia methods significantly impact mean arterial pressure, systolic and diastolic blood pressure, arterial blood pH and blood viscosity, but what kind of anesthesia method is more suitable for preparation of animal model does not reach an agreement. OBJECTIVE: To compare the effectiveness of local anesthesia and general anesthesia for preparation of rabbit intervertebral disc degeneration model in procession of percutaneous acupuncture rotary cut. METHODS:A total of 48 New Zealand white rabbits were randomly divided into the local anesthesia group and the general anesthesia group. The local anesthesia group was anesthetized with 0.5% lidocaine. The general anesthesia group was intraperitoneally injected with 3% sodium pentobarbital (30 mg/kg). In the two groups, models of intervertebral disc degeneration were established by minimally invasive percutaneous acupuncture rotary cut at L4/5, L5/6 levels. Modeling time was observed in both groups. Disc degeneration was evaluated by the gross observation, MRI detection, and histopathological examination at 4, 8, 12, and 16 weeks after modeling. RESULTS AND CONCLUSION: (1) General observation: the nucleus pulposus gradually with darker color and elasticity reduced in two groups after modeling. (2) MRI T2-weighted image results showed no significant changes in disc signal intensity at early stage, but the signal strength showed a decreasing trend with time. (3) Disc degeneration was evaluated according to Pfirrmann grading. Disc degeneration was gradually increased with time in two groups (P 0.05). (4) Masson staining showed that different degrees of annulus irregular arrangement appear in the two groups 8 and 12 weeks after modeling, but the structure was still intact. 16 weeks later, annulus disorganized, or even breakage in the two groups; no significant difference was detected between the two groups. (5) Safranin O staining showed that nucleus pulposus cells were not significantly reduced in the two groups at 4 weeks, but were significantly reduced at 16 weeks. (6) The average modeling time of local anesthesia group [(15.24±2.67) minutes] was significantly shorter than that in the general anesthesia group [(25.64±6.85) minutes] (P < 0.05). These results indicate that intervertebral disc degeneration model can be successfully established by minimally invasive percutaneous acupuncture rotary cut by using local anesthesia and general anesthesia; however, local anesthesia has a shorter operating time and simple anesthetic procedure. The efficacy was identical between the two methods.

Effect of Bushen Huoxue Tongluo Capsule on miR-27a in the synovium and chondrocytes in Sprague-Dawley rats with osteoarthritis / 中国组织工程研究

BACKGROUND: Bushen Huoxue Tongluo Capsule, a well-known traditional prescription, exhibits remarkable treatment outcomes in knee osteoarthritis, but the underlying mechanism is not fully understood. OBJECTIVE: To explore the regulatory effect of Bushen Huoxue Tongluo Capsule on miR-27a of synovial cells in Sprague-Dawley rats with knee osteoarthritis. METHODS: The model of knee osteoarthritis was established in 120 Sprague-Dawley rats using Hulth method, and then these rat models were randomized into four groups, followed by given75 mg/kg normal saline (blank control group), 34 (low-dose group), 75 (medium-dose group) and 140 (high-dose group) mg/kg Bushen Huoxue Tongluo Capsule aqueous solvent via gavage according to body mass, respectively, twice daily, for consecutive 2 courses (15 days as a course) with 2 days in between. The synovium and cartilages were removed after each course, and the expression of miR-27a in the knee joint synovium and chondrocytes was determined by real-time quantitative PCR. RESULTS AND CONCLUSION: Gross observation found that the degeneration of synovium and cartilage in the Bushen Huoxue Tongluo Capsule groups was milder than that in the blank control group. Real-time PCR revealed that there was an increasing trend in the expression of miRNA-27a in the synovium and cartilage in each group, especially in the high-dose group, and there was a significant difference between groups (P < 0.05). Our findings show that Bushen Huoxue Tongluo Capsule can suppress the reduced expression of miRNA-27a in the synovium and chondrocytes in rats with knee osteoarthritis, further alleviating knee degeneration and protecting the knee from damage.

Analysis on drug resistance of 196 strains of Mycobacterium tuberculosis in patients with HIV/AIDS complicating tubercolosis / 重庆医学

Objective To study the drug resistance situation of Mycobacterium tuberculosis strains isolated from the patients with HIV/AIDS complicating tuberculosis (TB).Methods One hundred and ninety-six inpatients with AIDS complicating TB in our hospital from January 2012 to December 2015 were collected.The drug resistance situation of Mycobacterium tuberculosis strains cultured from sputum/tissue fluid/fiber liquid and pathological tissue submitted samples in the patients with AIDS complicating active TB was analyzed.The BACTEC MGIT960 systems was adopted to perform the bacterial identification.The drug sensitivity test was conducted by using 960 culture testing system.Results One hundred and ninety-six strains of Mycobacterium tuberculosis were cultured,the total drug resistance rate of Mycobacterium tuberculosis was 26.02％.These cases were divided into the ＞100/μL and ≤100/μL groups according to different CD4+ T cells count.The drug resistance situation of Mycobacterium tuberculosis to 4 kinds of first line anti-TB drugs were compared between these two groups.The comparison results found that the resistance rate of Mycobacterium tuberculosis strains to the first line anti-TB drugs had no statistical difference between the two groups (P＞0.05).The resistance rates of first line anti-TB drugs from high to low were isoniazid,rifampicin,streptomycin and ethambutol.The drug resistance rate of Mycobacterium tuberculosis strains to 5 types of drug resistant TB had no statistical difference(P＞0.05).Conclusion The drug resistance rate in the initial patients with HIV complicating TB is consistent with the average initial drugresistance level of TB patients in our country.The TB drug resistance rate in the patients with HIV complicating TB has no correlation with CD4+ T cells count.

Zero-P anterior cervical fixation system for multilevel cervical myelopathy / 中国组织工程研究

BACKGROUND: Zero-P anterior cervical fixation system is an important means for treatment of cervical myelopathy in recent years. Zero-P fixation system can achieve effective decompression of spinal cord and nerve, and restore cervical curvature, with good stability.OBJECTIVE: To investigate the biocompatibility of Zero-P for multilevel cervical myelopathy.METHODS: Totally 62 patients with multilevel cervical disease who were treated by anterior cervical discectomy and fusion with Zero-P were selected, including double segments in 47 cases, three segments in 13 cases and four segments in 2 cases. Clinical efficacy was evaluated by pain visual analogue scale score, Japanese Orthopedic Association score, neck disability index score at postoperative 1, 6, 12, 24 months. Simultaneously, incidence of postoperative complications, cervical intervertebral space height and Cobb angle changes were observed.RESULTS AND CONCLUSION: (1) 54 patients were followed 2 years after treatment by clinic or telephone. Mean operative time was (102.00±32.41) minutes; average blood loss was (62.45±18.36) mL. (2) Two patients affected mild throat discomfort at one day after operation, and the symptoms went to lift six days after. The remaining patients did not experience any other complications. (3) Visual analogue scale score and neck disability index score at 1, 6, 12 and 24 months after operation were all less than preoperatively (P < 0.05); Japanese Orthopedic Association scores were higher than preoperatively (P < 0.05); the Cobb angle was improved significantly and greater than before treatment (P < 0.05).Postoperative cervical intervertebral space height was significantly higher than preoperatively (P < 0.05). Japanese Orthopedic Association results showed that among 54 patients, excellent was found in 34 cases, good in 14 cases,average in 6 cases and poor in 0 cases; the excellent and good rate was 89%. (4) Zero-P has a good clinical efficacy for multilevel cervical myelopathy, relieves symptoms significantly and restores cervical curvature and height, and has a good biocompatibility.

Effects of Yaotu Granules on nuclear factor-kappaB signaling pathway in human nucleus pulposus cells / 中国组织工程研究

BACKGROUND:Nuclear factor-κB (NF-κB) is highly expressed in the degenerated intervertebral disc. The Herbal Compound formula Yaotu Granules for lumbar disc herniation has achieved good clinical results;however, the underlying mechanisms remain poorly understood. OBJECTIVE:To explore mechanisms underlying the effects of the drug serum of Yaotu Granules on NF-κB signaling pathway. METHODS:Eight New Zealand white rabbits were randomly divided into normal saline group, low-, middle-, and high-dose drug groups. Normal saline, 10, 20, and 40 mL of water decoction of Yaotu Granules were administered intragastrical y in the normal saline, low-, middle-, and high-dose drug groups for 7 days, respectively. The drug serum of Yaotu Granules was prepared using serum samples obtained from the common carotid artery at 2 hours after the last administration. Human nucleus pulposus cel s were maintained in DMEM containing 10%(v/v) drug serum for 24 hours. RESULTS AND CONCLUSION:The expression levels of interleukin (IL)-8, tumor necrosis factor (TNF)-α, p-P65, P50, IκB-α(the NF-κB inhibitor), and IκB kinaseβ(IKK-β) mRNA in human nucleus pulposus cel s in three drug serum groups were significantly decreased compared with the normal saline group;in addition, a similar change in the expression levels of p-P65, P50, IκB-α, and IKK-βprotein between groups were detected. Noticeably, the expression levels of IL-8, TNF-α, p-P65, P50, IκB-α, and IKK-βmRNA, as wel as the expression levels of p-P65, P50, IκB-α, and IKK-βprotein were gradual y decreased with the increased drug dose. Al these results suggest that the drug serum of Yaotu Granules can protect nucleus pulposus cel s and delay their degeneration through blocking NF-κB signaling pathway.

AIDS patients with opportunistic infections in Sichuan province：clinical analysis of 1 465 cases / 重庆医学

Objective To study clinical characters and prognosis of AIDS patients with opportunistic infections in Sichuan prov-ince .Methods We performed an retrospective analysis of 1 465 AIDS patients with opportunistic infections who were admitted into the transmitted disease hospital of Chengdu in recent 10 years .Results The overall mortality during hospitalization was 15 .49% . The leading cause of death was respiratory failure due to pneumonia (n=150 ,74 .62% ) or meningitis(n=47 ,23 .86% ) .Descending rank order of common opportunistic infections were respiratory tract ,the mouth swallows ,central nervous system ,gastrointestinal tract skin ,blood system .Descending rank order of common sites of infections were respiratory tract ,oropharynx ,central nervous system ,gastrointestinal tract skin ,reproductive tract .The multiple infections are common :more than 50% of patients suffering from two or more infections(50 .77% ) .Complexity of infection sites :42 .18% dual infection sites and 17 .20% three or more infec-tion sites .Conclusion The opportunistic infections disease spectrum of AIDS in Sichuan area has its own characteristics :multiple infections and multiple infections sites are both common ,death risks are high ,conditions of patients are severe generally ,and respir-atory failure is the main cause of death .

Clinical study of Ruiyun procedure for hemorrhoids combined with Xiaozhiling injections in treatment of hemorrhoids complicated with human immunodeficiency virus infection / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To explore the efficacy of Ruiyun procedure for hemorrhoids (RPH) combined with Xiaozhiling injection in the treatment of hemorrhoids complicated with human immunodeficiency virus (HIV) infection and its influence on cellular immune function.</p><p><b>METHODS</b>Clinical data of 76 hemorrhoid patients, including 36 positive HIV and 40 negative HIV, undergoing RPH combined with Xiaozhiling injections in our center from January 2010 to December 2012 were retrospectively analyzed. Clinical efficacy and cellular immune function preoperative day 1, postoperative day 7, 30 were compared between positive and negative groups.</p><p><b>RESULTS</b>Recurrence rates of positive group and negative group postoperative 6 months were 22.2% (8/36) and 22.5% (9/40), postoperative 1 year were 30.6% (11/36) and 30.0% (12/40) without significant differences (all P>0.05). Morbidity of postoperative complication was also not significantly different between two groups (P>0.05). According to HIV classification, peripheral lymph cell ratio, CD4 count, CD4/CD8, white blood cell count and neutrophil ratio were not significantly different between preoperative day 1 and postoperative day 7 in both groups (all P>0.05). Decreasing velocity and amplitude of CD4 in both groups from high to low was HIV III, HIV II, HIV I, HIV-, while after 30 days the increase of CD4 from high to low was HIV-, HIV I, HIV II, HIV III, which were significantly different as compared to postoperative day 7 (all P<0.05).</p><p><b>CONCLUSIONS</b>RPH combined with Xiaozhiling injection in the treatment of hemorrhoids complicated with HIV infection is effective and safe. Postoperative inhibited cellular immune function can recover quickly.</p>

Immune responses to recombinant hepatitis B virus vaccine in human immunodeficiency virus-1-infected patients with different CD4+ T-lymphocyte / 中华传染病杂志

Objective To compare the difference of immune responses to hepatitis B virus (HBV) vaccine in human immunodeficiency virus (HIV)-1-infected patients with different CD4+ T-lymphocyte counts.Methods HIV-1-infected patients who visited clinic at the Public Health Clinical Center of Chengdu were enrolled and divided into two groups according to CD4+ T-lymphocytes counts.CD4+ T-lymphocytes ＜200/μL,which were and ≥200/μL.All patients were injected intramuscularly with 20 μg of HBV vaccine at month 0,1 and 6,respectively.Vaccination responses were measured at1 and 7 months after first dose.The serum anti-hepatitis B sarface antigen titers of ≥ 10 mIU/mL were considered positive.The serum anti-HBs positive rates was compared by Chi-square test and anti-HBs titers was compared by Mann-Whitney test.Results Ninety-five HIV-1-infected patients were finally enrolled into the analysis,with 55 patients in group of CD4+ T-lymphocytes ＜200/μL and 40 patients in group of CD4+ T-lymphocytes ≥200/μL.The serum anti-HBs positivity rates at 1 and 7 months after first dose in group of CD4+ T-lymphocytes ＜200/μL were 40.0％ (22/55) and 50.9％ (28/55),respectively,which were 47.5％ (19/40) and 75.0％ (30/40) in group of CD4+ Tlymphocytes ≥200/μL.There was no significant difference of serum anti-HBs positivity rates between two groups after 1 month (x2 =5.652,P=0.017).But a significant higher positivity rate was found in group of CD4+ T-lymphocytes ≥200/μL than in group ofCD4+ Tlymphocytes＜200/μL after 7 months (x2=0.531,P=0.466).The median titers of anti-HBs at 7 months were 10.44 mIU/mL in grouP of CD4+ T-lymphocytes ＜ 200/μL and 222.73 mIU/ml in group of CD4+ T-lymphocytes ≥200/μL.There was significant difference of anti-HBs titers between two groups (Z=-3.600,P＜0.01).No patients experienced side effects to HBV vaccine.Logistic regression model analysis indicated that only CD4+ Tlymphocytes was related to anti-HBs positivity after HBV vaccination.Conclusions HIV-1-infected patients with CD4+ T-lymphocytes ≥200/μL have higher serum anti-HBs positivity rates and higher anti HBs titers than patients with CD4+ T-lymphocytes ＜200/μL.However,patients with CD4+ T lymphocytes ＜200/μL also have relatively low immune response.Thus,HBV vaccination should be conducted in all these patients at high risk.

The diagnostic value of distortion products otoacoustic emissions in pseudo-anacousia / 中国基层医药

0.05).The DPOAE incidence in sensorineural hearing loss group was 18%,compared with normal group and pseudo-anacousia group,the DPOAE incidence were significantly lower(P

Application of Gukang Gold Far-Infrared Magnetic Therapeutic Paste to Treatment of Lateral Epicondylitis / 医疗卫生装备

Objective To apply Gukang gold far-infrared magnetic therapeutic paste to the treatment of lateral epicondylitis.Methods 64 cases of lateral epicondylitis patients were randomly divided into treatment group and control group,each with 32 ones.The subjects from the treatment group were treated with Gukang gold far-infrared magnetic therapeutic paste,and the remained with ibuprofen sustained-release capsules.The results after one and two courses were analyzed statistically.Results After two-course treatment,the patient from the treatment group had an higher improvement rate than the others from the control group,with a P value less than 0.05.The treatment course of the treatment group was shorter than that of the control group,with a P value less than 0.05.Conclusion When compared with ibuprofen sustainedrelease capsule,Gukang gold far-infrared magnetic therapeutic paste is more effective for the treatment of lateral epicondylitis.

Effect of kebimin decoction on level of Th1、Th2-typed cytokines in patients with allergic rhinitis / 中国免疫学杂志

Objective:To observe the effects of sera Th1/Th2-typed cytokines in patients allergic rhinitis (AR) who were treated with kebimin decoction(KD),and to study the therapeutic mechanism of KD in AR.Methods:60 patients of AR were randomly divided into two groups (treating group,TG and control group,CG). Patients in TG were treated with KD,while those in CG were treated with Xinfang Rhinitis Capsule before and after treatment,the levels of serum Th1-typed cytokines IFN-?、IL-2、IL-12 and Th2-typed cytokines IL-4、IL-5、IL-10 were determined by enzyme-linked immune sorbent assay (ELISA); and compared with 30 healthy controls.Results:The levels of serum Th2-typed cytokines IL-4、IL-5、IL-10 were higher and the levels of Th1-typed cytokines IFN-?、IL-2、IL-12 were lower in AR than that in healthy control ( P 05).Conclusion:Kebimin decoction is highly effective in treating allergic rhinitis by regulating the levels of Th1、Th2-typed cytokines,correcting the imbalance Th1/Th2-typed cytokines network.