Ectodermal Dysplasia 1, Anhidrotic ; diagnosis ; genetics ; Ectodysplasins ; genetics ; Heterozygote ; Humans ; Infant, Newborn ; Male ; Mutation, Missense

Objective: To investigate the dynamic expressions of tenascin-C (TN-C) at different phases of atherosclerosis in ApoE-/-mice. Methods: Our research included in 2 groups: ApoE-/- group, n=50 SPF male mice with ApoE-/- and Control group, n=50 wild male C57BL/6 mice. Atherosclerosis model was established by high fat diet in both groups. The mice were sacrificed at 4, 8, 16, 24 and 32 weeks, blood lipids were examined, pathologic changes of plaque were observed by microscope for quantitative analysis and TN-C expressions in atherosclerosis plaque were measured by immunohistochemistry. Results: Compared with Control group, ApoE-/- group had elevated blood levels of total cholesterol (TC) and low density lipoprotein cholesterol (LDL-C), both P<0.05. In ApoE-/- group, plaque area and the ratio of plaque area/lumen area were increasing upon prolonged modeling time, all P<0.05; TN-C expressions were increasing by progress of atherosclerosis, the highest TN-C expression was found at 32 weeks of modeling (0.49±0.07) which was higher than it was at 8 weeks (0.04±0.02), 16 weeks (0.12±0.03) and 24 weeks (0.21±0.04), all P<0.05. Conclusion: TN-C expression was increasing with plaque progress which might be related to the development of atherosclerosis and plaque instability.

Objective To investigate the cognitive, intelligent and cranial MRI status of 4 patients with myotonic dystrophy type 1 (DM1). Methods A pedigree of 4 patients,admitted to our hospital from September 2011 to December 2011 and diagnosed as having DM1, were chosen in our study; Chinese edition tests of Wechsler adult intelligence scale and Wechsler memory scale were used to test the intelligence of these patients; Montreal cognitive assessment (MoCA) and Neurobehavioral cognitive status examination (NCSE) were applied to evaluate the patients' cognitive function.All of the 4 patients received a cranial MRI scan. Results Except for 1 patient,the other 3 patients had low intelligent quotient (IQ) and memory quotient (MQ),and their verbal IQs were all lower than performance IQs.Three patients showed impaired visual space/executive skills,calculation,memory,and conceptual thinking abilities; 1 patient showed impaired sentence repetition and verbal fluency on MoCA,though his final scores were within normal range. Three patients had brain atrophy to some extent on MRI scan,prominently in the frontal lobe and temporal lobe; white matter lesions and thickened skull were founded in all 4 patients. Conclusion Multiple cognition disorders and brain involvements can be found in DM1 patients and deserve further investigation.

OBJECTIVETo prepare cytochrome (CYP)2E1-silenced hepatocytes by lentivirus-mediated RNA interference technology and to investigate the hepatotoxicity of trichloroethylene (TCE) in CYP2E1-silenced hepatocytes.

METHODSShort hairpin RNA fragments were designed and synthesized and were then ligated into the lentiviral vector; single colonies were screened; the plasmid was extracted after PCR and sequence identification and then transferred into L02 hepatocytes; the CYP2E1-silenced hepatocytes were selected; real-time quantitative PCR and Western blot were used to evaluate the interference effects. The obtained CYP2E1-silenced hepatocytes, as well as normal L02 hepatocytes, were treated with TCE (0, 0.25, 0.50, 1.00, 2.00, and 4.00 mmol/L). The cell viability and half maximal inhibitory concentration (IC50) of TCE were measured; the apoptotic rate of cells was measured by flow cytometry; the mRNA expression levels of apoptosis genes and oncogenes were measured by real-time quantitative PCR.

RESULTSThe IC50s of TCE for L02 hepatocytes and CYP2E1-silenced hepatocytes were 15.1 mmol/L and 23.6 mmol/L, respectively. The apoptotic rate increased as the dose of TCE rose in the two types of cells; the CYP2E1-silenced hepatocytes hada significantly lower apoptotic rate than L02 hepatocytes when they were exposed to 2.0 and 4.0 mmol/L TCE (P < 0.05 or P < 0.01). The mRNA expression level of bcl-2 (anti-apoptosis gene) in CYP2E1-silenced hepatocytes was 15% ∼ 60% higher than that in L02 hepatocytes (P < 0.01), while the mRNA expression levels of caspase-3 and caspase-9 (apoptosis genes) in CYP2E1-silenced hepatocytes were 30% ∼ 60% lower than those in L02 hepatocytes (P < 0.01). The mRNA expression level of p53 (cancer suppressor gene) in CYP2E1-silenced hepatocytes was 81 - 278% higher than that in L02 hepatocytes (P < 0.01), while the mRNA expression levels of c-fos and k-ras (oncogenes) in CYP2E1-silenced hepatocytes were 20-68% lower than those in L02 hepatocytes (P < 0.01).

CONCLUSIONCYP2E1-silenced cells can be successfully prepared by lentivirus-mediated RNA interference technology. Silencing CYP2E1 gene can reduce the hepatotoxicity of TCE and inhibit the expression of some apoptosis genes and oncogenes, suggesting that CYP2E1 gene plays an important role in TCE metabolism and is related to the hepatotoxicity of TCE.

Apoptosis ; drug effects ; genetics ; Cell Line ; Cell Survival ; drug effects ; genetics ; Cytochrome P-450 CYP2E1 ; genetics ; metabolism ; Genetic Vectors ; Hepatocytes ; drug effects ; metabolism ; Humans ; Lentivirus ; genetics ; RNA Interference ; Trichloroethylene ; toxicity

To construct the secretive prokaryotic shuttle expression plasmid pBCG-SP-HSP65, the signal peptide sequence of antigen 85B amplified from Bacillus Calmette-guérin (BCG) genome by PCR and the whole HSP65 DNA sequence of human M. tuberculosis obtained from the plasmid pCMV-MTHSP65 by PCR were cloned into the plasmid pBCG-2100 under the control of the promoter of Heat Shock Protein 70 (HSP70) from human M. tuberculosis. Recombinants were electroporated into Mycobacterial smegmatis and induced by heating. Results of the induced expression were detected by SDS-PAGE and the biological activity of the expressed protein was tested by Western-blot analysis. Results showed pBCG-SP-MTHSP65 was constructed successfully and confirmed by restriction endonuclease analysis, PCR detection and DNA sequencing analysis. After it was electroporated into Mycobacterial smegmatis and induced by heating, the percentage of expressed 65kD protein in Mycobacterial smegmatis detected by SDS-PAGE was 20% in total bacterial protein. But the percentage of expressed 65kD protein in recombibinant Mycobacterial smegmatis was up to 34.46% in total bacterial protein and 68.56% in the total protein of cell lysate supernants, Which demonstrated the recombinant HSP65 gene could express in recombinant with high efficiency and the expressed proteins were mainly soluble. Western-blot showed that the secretive proteins could specially combine with antibody against human M. tuberculosis HSP65. Orally, pBCG-SP-HSP65 was successfully constructed; HSP65 gene could express in Mycobacterial smegmatis with high efficiency via it. And the expressed proteins possess the biological activity. So it provids experimental evidence for the application of the recombinant Mycobacterial smegmatis and the development of the vaccine against tuberculosis.
Bacterial Proteins ; biosynthesis ; genetics ; Chaperonin 60 ; Chaperonins ; biosynthesis ; genetics ; Escherichia coli ; genetics ; metabolism ; Humans ; Mycobacterium smegmatis ; genetics ; metabolism ; Plasmids ; genetics ; Recombinant Fusion Proteins ; biosynthesis ; genetics

OBJECTIVETo investigate the structural changes inintestinal flora and metabolic changes in type 2 diabetic patients with obesity(BMI≥40 kg/m)by sequencing the 16S rRNA genes.

METHODSStool samples were collected from 4 diabetic patients before and after gastric bypass surgery for extraction of the total DNA. The diversity of the intestinal flora in the samples was investigated by 16S rRNA sequencing. After surgery, the changes in glucose and lipid metabolism were evaluated in the patients, and the changesin body mass index (BMI) and waist to hip ratio were assessed at 3 month intervals.

RESULTSAfter gastric bypass, the patient's BMI, waist to hip ratio, glucose metabolism and lipid metabolism gradually recovered the normal levels. The proportion of Bacteroidetesis increased and the proportions of Firmicutes and Proteobacteria decreased in the intestinal bacteria after the surgery.

CONCLUSIONGastric bypass surgery can effectively alleviate the condition of obese patients with type 2 diabetes and improve the composition of the intestinal flora.

Lung cancer， a malignancy with high incidence rate and mortality rate， is a major threat to human life and health. At present， the common methods for the treatment of lung cancer include surgical resection， radiotherapy， chemotherapy， targeted therapy， and immunotherapy， but these methods generally have the problems of severe toxic/side effect and high treatment cost. Traditional Chinese medicine（TCM） has a history of more than 2 000 years of application in China and has its unique advantages in the treatment of tumors. Modern pharmacological experiments have found that TCM can inhibit tumor growth， prolong patients' survival， and improve clinical symptoms and patients' quality of life by inducing tumor cell apoptosis， inhibiting tumor angiogenesis， and reducing tumor cell drug resistance. Apoptosis is a process of spontaneous programmed cell death， which is closely related to the occurrence and development of the tumor. Studies have shown that many Chinese medicines can inhibit the development of lung cancer by inducing apoptosis. This study searched， analyzed， and summarized the available papers on the mechanism of TCM in the treatment of lung cancer by inducing apoptosis. It is found that Chinese medicine induces lung cancer cell apoptosis mainly by regulating apoptosis-related factors and apoptosis-related signaling pathways ［inhibitor of apoptosis proteins （IAPs）， B cell lymphoma-2 （Bcl-2）， p53 protein， the second mitochondria-derived activator of caspase （SMAC）/direct IAP-binding protein with low isoelectric point （DIABLO）， extrinsic apoptotic pathway， endogenous mitochondrial pathway， Janus kinase （JAK）/signal transducer and activator of transcription （STAT） signaling pathway， mitogen-activated protein kinase （MAPK） signaling pathway， and phosphoinositide 3-kinase （PI3K）/protein kinase B （Akt） signaling pathway. In addition， the Wnt/β-catenin/survivin signaling pathway and the Notch signaling pathway also play an important role in inducing apoptosis.

OBJECTIVE: To explore clinical effects of ultrasound-guided minimal traverse-cross technique repair for acute closed Achilles tendon ruptures. METHODS: From January 2015 to March 2017, 20 patients with acute closed Achilles tendon rupture were treated by minimal traverse-cross technique repair with ultrasound guided. Among them, including 13 males and 7 females, aged from 28 to 49 years old with an average of(31.3 ±4.5) years old. All patients were single side injury. Fifteen patients on the left side and 5 patients were on the right side. The time from injury to operation ranged from 1 to 5 days with an average of (2.5±0.7) days. Operative time, postoperative complications were observed, and AOFAS score before and after operation at 12 months were compared. RESULTS: All patients were followed up for 12 to 27 months with an average of(15.2±4.9) months. Operative time ranged from 33 to 65 min with an average of(43.7±5.6) min. Incision of one patient were continued oozing and improved after changing dressings, other patients were healed at stage I. No sural nerve irritation symptoms and palindromic rapture of heel tendon occurred. AOFAS score was improved from 65.2±7.4 before operation to 97.7±4.7 after operation at 12 months (t=22.5, <0.01); 18 patients got excellent results and 2 good. CONCLUSIONS Ultrasound-guided minimal traverse-cross technique repair for acute closed Achilles tendon ruptures, which promise minimal incision, protect sural nerve, ensure quality of tendon anastomosis and fixation, and is a ideal method for repairing acute closed Achilles tendon ruptures.
Achilles Tendon ; Adult ; Female ; Humans ; Male ; Middle Aged ; Rupture ; Sural Nerve ; Sutures ; Tendon Injuries ; Treatment Outcome

Fermented Chinese medicine has long been used. Amid the advance for preservation of experience, the connotation of fermented Chinese medicine has been enriched and improved. However, fermented Chinese medicine prescriptions generally contain a lot of medicinals. The fermentation process is complicated and the conventional fermentation conditions fail to be strictly controlled. In addition, the judgment of the fermentation end point is highly subjective. As a result, quality of fermented Chinese medicine is of great difference among regions and unstable. At the moment, the quality standards of fermented Chinese medicine are generally outdated and different among regions, with simple quality control methods and lacking objective safe fermentation-specific evaluation indictors. It is difficult to comprehensively evaluate and control the quality of fermented medicine. These problems have aroused concern in the industry and also affected the clinical application of fermented Chinese medicine. This article summarized and analyzed the application, quality standards, and the modernization of fermentation technology and quality control methods of fermented Chinese medicine and proposed suggestions for improving the quality standards of the medicine, with a view to improving the overall quality of it.
Medicine, Chinese Traditional ; Reference Standards ; Quality Control ; Fermentation