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1.
Chinese Journal of Biotechnology ; (12): 1033-1045, 2018.
Article in Chinese | WPRIM | ID: wpr-687712

ABSTRACT

The high efficiency and stability of enzymes are the basis for industrial application. Hybrid enzyme suitable for industrial applications could be constructed by many molecular biology technologies including tandem fusion, domain insertion and post-translational protein conjugation. However, the low expression and activity of hybrid enzyme limit its application in industrial production, and multifunctional design of a specific protein domain has been becoming a new trend. With the advent of high-throughput sequencing, biologists are starting to wrestling with massive data sets. Besides, the concept of protein sectors and co-evolution provides novel insight into the relationship of protein structure and function. The residues-covariation of a protein sector displays preference, which imparts functional diversity to different enzymes in the same family. The covariation-residues in specific protein sectors can be located based on the analysis of massive data, and then these functional residues can be assembled in a new enzyme variant using the biotechnology of synthetic biology, thus completing the redesign of natural enzymes. This indicates a new stage of designing hybrid enzyme, as well as the new trend of protein design in the era of biological big data.

2.
Article in Chinese | WPRIM | ID: wpr-706805

ABSTRACT

Objective:To investigate the effect of miRNA-34b/c-5p on the expression of neurokinin 1 receptor-truncated(NK1R-Tr)in breast cancer and the effect of miRNA-34b/c-5p and NK1R-Tr on the migration and invasion abilities of breast cancer cells.Methods:Real-time fluorescence quantitative PCR(RT-PCR)was used to detect the expression of miRNA-34b/c-5p and NK1R-Tr in 50 breast can-cer specimens that were collected at Tianjin Medical University Cancer Institute&Hospital from February to May 2013.Western blot analysis was performed to detect the expression of miRNA-34b/c-5p and NK1R-Tr in breast cancer cell lines MDA-MB-231 and MCF-7. Scratch and Transwell assays were carried out to explore the effects of miRNA-34b/c-5p and NK1R-Tr on the migration and invasion abilities of MDA-MB-231 cells.Results:The expression of miRNA-34b/c-5p and NK1R-Tr in breast cancer tissues and cells were signifi-cantly negatively correlated.The relative expression of NK1R-Tr in breast cancer patients with lymph node metastasis was 5.75,and the relative expression of NK1R-Tr in patients with non-lymph node metastasis was 4.29.The relative expression was significantly dif-ferent between the two groups(P=0.026).Overexpression of miRNA-34b/c-5p and knockdown of NK1R-Tr could significantly inhibit the migration and invasion abilities of MDA-MB-231 cells(all P<0.001).Conclusions:The expression of miRNA-34b/c-5p and NK1R-Tr was significantly negatively correlated.MiRNA-34b/c-5p and NK1R-Tr might be potential therapeutic targets for inhibiting the invasion of breast cancer cells.

3.
Article in Chinese | WPRIM | ID: wpr-319949

ABSTRACT

<p><b>OBJECTIVE</b>To explore the effects of principal-subordinate acupoints combination on improving myocardial ischemia, and the gene regulatory pathways for the protection of myocardial ischemia.</p><p><b>METHODS</b>According to the random number table method, 70 SPF Wistar male rats were divided into a normal group, a model group, a LY294002 group, an insulin-like growth factors-1(IGF-1) group, a Neiguan group, an acupoint combination group and an acupoint combination + LY294002 group, 10 rats in each one. Rats in the normal group were injected with 0.9% NaCl solution, while rats in the remaining groups were treated with abdominal subcutaneous injection of isoroterenol hydrochloride to establish the rat model of myocardial ischemia. Rats in the LY294002 group and IGF-1 group were treated with injection of LY294002 solution and IGF-1 solution for 14 days. Rats in the Neiguan group were treated with electroacupuncture (EA) at "Neiguan" (PC 6) by using Han-200 EA apparatus for 10 min per treatment. Rats in the acupoint combination group were treated with EA at "Neiguan" (PC 6), "Zusanli" (ST 36) and "Guanyuan" (CV 4) by using Han-200 EA apparatus for 10 min per treatment. Rats in the acupoint combination + LY294002 group were treated with LY294002 solution for 14 days, and EA at "Neiguan" (PC 6), "Zusanli" (ST 36) and "Guanyuan" (CV 4) was given before model establishment, once a day for 21 days. EA pretreatment was given before model establishment in all acupuncture groups. The heart rate (HR) and ST segment voltage were detected before and after treatment; the myocardial pathological morphology was observed by HE staining; the expressions of P13K mRNA and Akt mRNA were tested.</p><p><b>RESULTS</b>After modeling, HR and ST segment voltage in all intervention groups were higher than those in the normal group (all P < 0.01); after the intervention, the HR and the ST segment voltage in the acupoint combination group, IGF-1 group and IGF-1 group were improved (P < 0.01, P < 0.05), which was more significant in the acupoint combination group and Neiguan group (all P < 0.01). As for the myocardial pathological morphology, obvious myocardial ischemia was observed in the model group, and that in the LY294002 group was the most serious, and that in the acupoint combination+ LY294002 group was moderate. After intervention, the myocardial pathological damage in the IGF-1 group, Neiguan group and acupoint combination group was significant improved, which was more significant in the IGF-1 group and acupoint combination group. As for the expression of PI3K mRNA and Akt mRNA, compared with normal group, the expression of PI3K mRNA was increased in the remaining groups after modeling (P < 0.01, P < 0.05), which was more significant in the IGF-1 group and acupoint combination group (all P < 0. 01). The expression of Akt mRNA in the LY294002 group and acupoint combination + LY294002 group was reduced (P < 0. 01, P < 0.05), while that in the remaining groups was increased (P < 0.01, P < 0.05), which was more significant in the IGF-1 group and acupoint combination group (all P < 0.01).</p><p><b>CONCLUSION</b>The principal-subordinate acupoints combination could improve heart rate and ST segment voltage in rats with chronic myocardial ischemia, reduce myocardial pathological damage, which is superior to single selection of "Neiguan" (PC 6). The PI3K/Akt signaling pathway may be involved in the regulation mechanism of principal-subordinate acupoints combination for the protection of chronic myocardial ischemia.</p>


Subject(s)
Acupuncture Points , Acupuncture Therapy , Animals , Chronic Disease , Therapeutics , Electroacupuncture , Electrocardiography , Heart Rate , Humans , Insulin-Like Growth Factor I , Metabolism , Male , Myocardial Ischemia , Pathology , Therapeutics , Myocardium , Pathology , Phosphatidylinositol 3-Kinases , Genetics , Metabolism , Proto-Oncogene Proteins c-akt , Genetics , Metabolism , Rats , Rats, Wistar
4.
Article in Chinese | WPRIM | ID: wpr-269755

ABSTRACT

<p><b>OBJECTIVE</b>To compare the clinical effects between ginger-partition moxibustion combined with glucocorticoid and simple oral glucocorticoid for thyreoitis at subacute stage.</p><p><b>METHODS</b>Eighty-one patients were randomly divided into an observation group (41 cases) and a control group (40 cases). In the observation group, ginger-partition moxibustion and hormone were applied. Moxa cones were used at local ashi points, Zusanli (ST 36), Guanyuan (CV 4) and Qihai (CV 6), six cones every point, once every other day and three times a week. Besides, 24 mg methylprednisolone tablets were adopted orally every day, and in two weeks the dose was 16 mg/d, in four weeks 8 mg/d, in six weeks 4 mg/d; all the patients were observed for 8 week. In the control group, simple methylprednisolone was prescribed orally, and the dose, the usage and treatment time were the same as those in the observation group. Erythrocyte sedimentation rate (ESR), triiodothyronine (T3), thyroxin (T4) and ultra-sensitive thyroid stimulating hormone (TSH) before and after treatment were observed in the two groups, as well as fever, the pain of thyroid gland, the regression time of swelling and adverse reaction.</p><p><b>RESULTS</b>The time of thyroid gland pain relieved of the observation group was earlier than that of the control group [(3.07 ± 0.78) days vs (3.62 ± 0.92) days, P < 0.05]. After treatment, T3, T4 and ESR were declined apparently (all P < 0.01), and TSH was obviously increased in the two groups (both P < 0.01). After 2-week treatment, ESR in the observation group was lower than that in the control group (P < 0.05). After 4-week treatment, T3, T4, TSH and ESR in the observation group were better than those in the control group (all P < 0.05). After 8 weeks, all indices in the observation group were superior to those in the control group, without statistical significance between the two groups (all P > 0.05). The effects of the observation group in 2 weeks, 4 weeks and 8 weeks were better than those in the control group (all P < 0.05). The cured course was shorter in the observation group than that in the control group (P < 0.05). The adverse reaction rate in the observation group was lower than that in the control group [4.9% (2/41) vs 22.5% (9/40), P < 0.05]. Three months later after treatment, the cured patients were followed. There was no recrudescence in the observation group and three patients caught the disease again in the control group, but there was no statistical significance between the two groups (P > 0.05).</p><p><b>CONCLUSION</b>Ginger-partition moxibustion combined with glucocorticoid achieves better effect than simple oral glucocorticoid for thyreoitis at subacute stage, and the adverse reaction is less, which presents the clinical advantages of the integration of Chinese and western medicine.</p>


Subject(s)
Adult , Female , Ginger , Chemistry , Glucocorticoids , Humans , Male , Middle Aged , Moxibustion , Thyroid Gland , Metabolism , Thyroid Hormones , Metabolism , Thyroiditis , Drug Therapy , Pathology , Therapeutics , Thyrotropin , Metabolism , Young Adult
5.
Article in Chinese | WPRIM | ID: wpr-490306

ABSTRACT

Objective To observe the clinical efficacy of bloodletting cupping at Dazhui (GV14) plus scraping in treating hordeolum. Method Forty-four patients with hordeolum were randomized into a control group and a treatment group, 22 cases in each group. The control group was intervened by bloodletting cupping at Dazhui, while the treatment group was by bloodletting cupping at Dazhui plus scraping. The size of the infection on eyelid was observed before and after treatment, and the recovery rate was calculated. Result The total effective rate was 81.8% in the control group versus 95.5% in the treatment group, and the total effective rate in the treatment group was higher than that in the control group (P<0.05); the sizes of the eyelid infection were significantly reduced in both groups after treatment (P<0.05), and the reduction in the treatment group was more significant than that in the control group (P<0.05). Conclusion Bloodletting cupping at Dazhui plus scraping can effectively improve the symptoms, reduce infection size, and enhance the clinical efficacy in treating hordeolum.

6.
Article in Chinese | WPRIM | ID: wpr-490197

ABSTRACT

Objective To observe the clinical efficacy of thermal electroacupuncture therapy in treating lumbodorsal myofascitis. Method Sixty-two patients with lumbodorsal myofascitis were randomized into a treatment group of 30 cases and a control group of 32 cases. The treatment group was intervened by thermal electroacupuncture therapy, while the control group was by orally taking Meloxicam tablets. The Visual Analogue Scale (VAS) was observed before and after treatment, and the clinical efficacies were compared.Result The total effective rate was 100.0% in the treatment group versus 78.1% in the control group, and the difference was statistically significant (P<0.05). The VAS scores were significantly changed after treatment in both groups (P<0.05). After treatment, the VAS score in the treatment group was significantly different from that in the control group (P<0.05).Conclusion Thermal electroacupuncture therapy is an effective approach in treating lumbodorsal myofascitis.

7.
Article in Chinese | WPRIM | ID: wpr-352617

ABSTRACT

<p><b>OBJECTIVE</b>To observe the effects of electroacupuncture (EA) at different acupoints on apoptosis-related serum and expression of microRNA (miRNA) in rats with myocardial ischemia, so as to explore its mechanism of action.</p><p><b>METHODS</b>A total of 48 male Wistar rats were randomly divided into a normal group, a model group, a Neiguan group and a acupoint compatibility group, 12 rats in each group. Isoprenaline hydrochloride (ISO) with a daily dose of 2 mg/kg was subcutaneously injected for 14 days to establish the myocardial ischemia model in the model group, Neiguan group and acupoint compatibility group. Rats in the normal group were subcutaneously injected with an equal volume of normal saline. After modeling, rats in the Neiguan group were treated with EA at "Neiguan" (PC 6), while rats in the acupoint compatibility group were treated with EA at "Guanyuan" (CV 4), "Zusanli" (ST 36) and "Neiguan" (PC 6). Rats in the normal group and model group were treated with immobilization, once day for 21 days. The contents of creatine kinase-MB (CK-MB), vascular cell adhesion molecule-1 (VCAM-1) and endothelin-1 (ET-1) in serum were detected by enzyme-linked immunosorbent assay (ELISA); apoptosis index (AI) of myocardial cells was detected by TUNEL method; the expressions of miRNA-1, miRNA-133, miRNA-208 and miRNA-499 were detected by real-time PCR method.</p><p><b>RESULTS</b>Compared with the normal group, the serum CK-MB, VCAM-1 and ET-1 were significantly increased in the model group, Neiguan group and acupoint compatibility group (all P < 0.01), and the apoptosis index was significantly increased (all P < 0.01). The CK-MB, VCAM-1 and ET-1 in the Neiguan group and acupoint compatibility group were significantly lower than those in the model group (all P < 0.01); the AI was reduced, which was more significant in the acupoint compatibility group (P < 0.05). Compared with the normal group, the expression of miRNA-133 was reduced (P < 0.01) and those of miRNA-208, miRNA-1 and miRNA-499 were significantly increased in the model group (all P < 0.01). Compared with the model group, the expression of miRNA-133 was increased (both P < 0.01) and that of miRNA-208, miRNA-1 and miRNA-499 were significantly reduced (all P < 0.01) in the Neiguan group and acupoint compatibility group. Compared with the Neiguan group, the expression of miRNA-133 was increased (P < 0.01) and those of miRNA-208, miRNA-1 and miRNA-499 were significantly reduced in the acupoint compatibility group (P < 0.01, P < 0.05).</p><p><b>CONCLUSION</b>EA at acupoints, especially acupoint compatibility group, could effectively prevent and treat myocardial ischemia, and the protective effect is possibly correlated to the double regulation on increasing the expression of miRNA-133 and inhibiting the expression of miRNA-1, miRNA-208, miRNA-499.</p>


Subject(s)
Acupuncture Points , Animals , Apoptosis , Disease Models, Animal , Electroacupuncture , Endothelin-1 , Genetics , Metabolism , Humans , Male , MicroRNAs , Genetics , Metabolism , Myocardial Ischemia , Genetics , Metabolism , Therapeutics , Myocytes, Cardiac , Metabolism , Rats , Rats, Wistar
8.
Article in Chinese | WPRIM | ID: wpr-458110

ABSTRACT

Objective: To observe the therapeutic effect of acupuncture plus psychological intervention for postpartum depression. Methods: By random number table, 85 patients with postpartum depression were divided into a treatment group and a control group. Forty-three cases in the treatment group were treated by acupuncture plus psychological intervention, once every day, five sessions per week, and rest at weekend. Forty-two cases in the control group were treated by oral administration of Fluoxetine Hydrochloride, 20 mg, once per day. The two groups were treated continuously for six weeks. The change of the score in Hamilton depression scale (HAMD) was observed and the therapeutic effect was summarized. Results:The total effective rate was 90.7%in the treatment group and 90.5% in the control group. The difference in the total effective rate between two groups was not statistically significant (P>0.05). In the intra-group comparisons of HAMD scores two, four and six weeks after treatment in both groups with those before treatment, the differences were statistically significant (all P0.05). Regarding the adverse events, 5 cases had nausea, 3 cases had dizziness, and 6 cases had poor appetite in the control group; no obvious adverse events happened in the treatment group. Conclusion:Acupuncture plus psychological intervention for postpartum depression is as same as oral administration of Fluoxetine Hydrochloride in therapeutic effects, but it does not have adverse reaction.

9.
Chinese Journal of Biotechnology ; (12): 422-433, 2013.
Article in Chinese | WPRIM | ID: wpr-233233

ABSTRACT

Molecular engineering of cellulases can improve enzymatic activity and efficiency. Recently, the Carbohydrate-Active enZYmes Database (CAZy), including glycoside hydrolase (GH) families, has been established with the development of Omics and structural measurement technologies. Molecular engineering based on GH families can obviously decrease the probing space of target sequences and structures, and increase the odds of experimental success. Besides, the study of cellulase active-site architecture paves the way toward the explanation of catalytic mechanism. This review focuses on the main GH families and the latest progresses in molecular engineering of catalytic domain. Based on the combination of analysis of a large amount of data in the same GH family and their conservative active-site architecture information, rational design will be an important direction for molecular engineering and promote the rapid development of the conversion of biomass.


Subject(s)
Catalytic Domain , Genetics , Cellulase , Chemistry , Genetics , Directed Molecular Evolution , Methods , Evolution, Molecular , Glycoside Hydrolases , Chemistry , Genetics , Protein Engineering , Methods
10.
Chinese Journal of Orthopaedics ; (12): 1122-1127, 2011.
Article in Chinese | WPRIM | ID: wpr-422614

ABSTRACT

ObjectiveTo evaluate the clinical outcomes of anterior lumbar discectomy and interbody fusion with cage under laparoscopic assistant.MethodsFrom January 2006 to June 2009,37 cases with degenerative low back pain were entered the study,including 22 males and 15 females with an average age of 43.7 years(range,16-55).The responsible discs were determined according to the three dimensional computed tomography of artery and vein angiography of anterior lumbosacral spine and discography,including L5S1 in 21 cases,L4-5 in 11,L3-4 in 2,L2-3 in 2,and L1-2 in 1.All cases underwent anterior lumbar discectomy and interbody fusion with cage under laparoscopic.ResultsThe operation time was 100 min in average (range,60-140),the blood loss was 120 ml in average(range,50-300).There was no case with severe complications of retrograde ejaculation and injury of great vessels or nerves.Delayed intestinal obstruction was discovered in two intraperitoneal route patients.The average follow-up time was 18.7 months(range,6-35).According to the back pain grading criteria of Chinese Medical Association Orthopedics Society of Spine Group,the results were excellent in 23 patients,good in 11,and fair in 3.The interbody fusion was obtained in 3 months later in 23 cases and 6 months later in 12 cases.Cage subsidence occurred in 2 cases in 6months after operation,in which the height loss of intervertebral space was 1.3 mm and 1.9 mm,but no obvious symptoms of discomfort.No fixation displacement or loosening occurred.ConclusionThe anterior discectomy and interbody fusion by internal fixation with laparoscopic technique is feasible with low complications rate,less trauma and shorter bedtime.Postoperative ileus by abdominal approach is relatively common.The surgeons experience and the anatomy of artery and vein of anterior lumbosacral spine should be considered before the choice of surgical approach.

11.
Chinese Journal of Biotechnology ; (12): 864-869, 2010.
Article in Chinese | WPRIM | ID: wpr-292197

ABSTRACT

Biofuels and bio-based chemicals from lignocellulosic biomass are sustainable, making them alternatives to petroleum-derived fuels and chemicals to address the challenges of the shortage of crude oil supply and climate change resulted from the overconsumption of petroleum-based products, particularly in China. However, high cost in liberating sugars from lignocellulosic biomass is still the bottleneck of the commercialization of biofuels and bio-based chemicals. In this article, the major components of cellulases and their synergistic role in the hydrolysis of pre-treated biomass is reviewed, followed by how to evaluate the enzymatic hydrolysis. With the elucidation of the underlying mechanism of the conformations of the enzyme molecules and their effectiveness in attacking cellulose substrate, more efficient enzymes are expected to be developed. Using the high production strain Penicillium decumbens, the on-site production of cellulases for cellulose ethanol production is discussed.


Subject(s)
Biofuels , Biomass , Biotechnology , Methods , Biotransformation , Cellulase , Hydrolysis , Industrial Microbiology , Methods , Lignin , Chemistry , Metabolism
12.
Chinese Journal of Biotechnology ; (12): 1838-1843, 2009.
Article in Chinese | WPRIM | ID: wpr-336298

ABSTRACT

There is a great diversity for cellulolytic microbes in nature and the strategies they use to digest cellulose. In addition to the cultured cellulolytic microbes, there are still a great number of microbes being not readily culturable in natural environments, which may represent great potential for identifying novel cellulases and their encoding genes. The rise of metagenomics and metaproteomics provides essential technologic tools to dig up these resources and significant progress has been made so far. This review gives an insight into some relative results that have arisen from the meta-genomic or proteomic analysis of definitive uncultured microbe communities. Their potential role in elucidating the process and mechanisms of cellulose degradation in natural environment from the point of "community system microbiology" is also discussed.


Subject(s)
Bacteria , Genetics , Cellulases , Genetics , Metabolism , Cellulose , Metabolism , Cloning, Molecular , Environmental Microbiology , Genome, Bacterial , Metagenomics
13.
Article in Chinese | WPRIM | ID: wpr-406013

ABSTRACT

BACKGROUND:Techniques of antedor cervical discectomy and interbody fusion have achieved good clinical outcomes since Robinson and Smith first introduced it in 1955.Currently,microendoscopic approaches have resulted in good clinical effectiveness in treating cervical spondylosis.Cage has also been used widely in the clinic.OBJECTIVE:To find a minimally invasive method that uses microendoscopic technique and Cages to treat cervical spondylosis by anterior approach.DESIGN,TIME AND SETTING:A retrospective case analysis was performed at the Department of Spinal Surgery,First Affiliated Hospital of University of South China between January 2003 and July 2008.PARTICIPANTS:A total of 25 patients with cervical spondylosis,15 males and 10 females,averaging 39 years of age (range,33-58 years old),who received treatments at the Department of Spinal Surgery,First Affiliated Hospital of University of South China were included in this study.Each of them had unilateral limb pain,numbness or weakness in varying degrees.MRI image showed one-level cervical disc degeneration in 20 patients and two levels in 5 patients.METHODS:Twenty-five patients underwent anterior cervical microendoscopic discectomy and interbody fusion using carbon fiber reinforced polymer (CFRP) Cage.Surgical techniques,neurological outcomes and complications were retrospectively analyzed.The postoperative treatments were just like the conventional approaches.MAIN OUTCOME MEASURES:Improvements in neurological symptoms were assessed using the Japanese Orthopedic Association (JOA) grading system.Postoperative imaging data including lordosis and fusion condition were examined.RESULTS:All patients received treatments successfully.The mean surgical time was 140 minutes (range,80 to 225 minutes).The hospital stay averaged 7 days (range,6 to 10 days).Patients' neurological statuses improved to varying degrees according to the JOA grading system:7.5 pre-operative (range,4 to 9 points) to 12 post-operative (range,8 to 16 points),with the mean improvement rate of 45.6%.All cases were followed up and the mean follow-up period was 18 months (range,6 to 27 months).Postoperative images showed solid bone fusion without implant migration.No vascular injury and infection occurred.Graft collapse was found in 2 patients,and transient hoarse voice in 1 patient.No other complications were found.CONCLUSION:Anterior microendoscopic cervical discectomy and interbody fusion with CFRP Cage in treatment of cervical spondylosis have the advantages of minimal invasion,sufficient decompression,and satisfactory bone fusion,resulting in a significant relief of patients' preoperative clinical symptoms.

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