Fibroblast growth factor -21 (FGF-21) is a recently discovered metabolic regulation factor, regulating glucose and lipid metabolism and increasing insulin sensitivity. FGF-21 is expected to be a potential anti-diabetic drug. Expression of FGF-21 as inclusion bodies has advantages for high yield and purity, but the bioactivity of the protein is almost totally lost after denature and renature. That is why FGF-21 is currently expressed in soluble form. As a result, the yield is considerably low. In this study, we used SUMO vector to express SUMO-human FGF-21 (SUMO-hFGF-21) in form of inclusion body. We optimized the culture conditions to increase the yield of the bioactive human fibroblast growth factor-21. We applied the hollow fiber membrane filtration column to enrich the bacteria, wash, denature and renature inclusion bodies. After affinity and gel filtration chromatography, we examined the hypoglycemic activity of FGF-21 by the glucose uptake assay in HepG2 cells. We also detected the blood glucose concentration of type 2 diabetic db/db model mice after short or long-term treatment. The results show that the yield of ihFGF-21 was 4 times higher than that of shFGF-21. The yield was 20 mg/L for ihFGF-21 vs. 6 mg/L for shFGF-21. The purity of ihFGF-21 was above 95%, while shFGF-21 was 90%. Compared with the traditional method of extracting inclusion bodies, the production cycle was about three times shortened by application of hollow fiber membrane filtration column technology, but the bioactivity did not significantly differ. This method provides an efficient and cost-effective strategy to the pilot and industrial production of hFGF-21.
Animals ; Bacteria ; metabolism ; Diabetes Mellitus, Experimental ; drug therapy ; Disease Models, Animal ; Fibroblast Growth Factors ; biosynthesis ; Genetic Vectors ; Glucose ; metabolism ; Hep G2 Cells ; Humans ; Hypoglycemic Agents ; isolation & purification ; Inclusion Bodies ; metabolism ; Mice ; Recombinant Fusion Proteins ; biosynthesis ; Small Ubiquitin-Related Modifier Proteins ; biosynthesis

BACKGROUND:There is a great dispute on the choice of repair materials for ossicular chain damage.
OBJECTIVE:To explore the effects of new hydroxyapatite-bone morphogenetic protein ossicular prosthesis versus al ogeneic cartilage ossicular prosthesis in patients with ossicular chain damage.
METHODS:Sixty patients with chronic otitis media were equal y assigned into a control group and a test group. Two groups of patients underwent tympanoplasty and ossicular chain reconstruction with al ogeneic cartilage ossicular prosthesis or hydroxyapatite-bone morphogenetic protein ossicular prosthesis, respectively. Twelve months after surgery, fol ow-up results were compared between two groups.
RESULTS AND CONCLUSION:Air conduction value and air-bone gap value were both improved significantly in the two groups after surgery (P<0.05), and especial y, the postoperative air conduction value was better in the test group than the control group (P<0.05). Implant detachment was found in three cases of the control group, whereas did not occur in the test group. Improved hearing was found in 28 cases in the test group with an improvement rate of 93%and 25 cases in the control group with an improvement rate of 83%. There was a significant difference in the hearing improvement between the two groups (P<0.05). Overal , two kinds of prostheses can share similar effects on the ossicular chain repair.
Subject headings:Tympanoplasty;Otitis Media;Bone Morphogenetic Proteins;Tissue Engineering

Background and purpose:The previous research has found that the prostate stromal cells derived from different prostate zones have distinct effect on prostate epithelial cells. We also revealed that LMO2 protein was highly expressed in PZ stromal cells (PZSCs) and prostate cancer associated fibroblasts (CAFs) compared with TZ stromal cells. This study investigated the effect of LMO2 protein in prostate stromal cells on proliferation and invasion of prostate cancer PC-3 cells and its mechanisms. Methods:Lentivirus overexpression vectors were used to establish LMO2-overexpressed prostate WPMY-1 stromal cell line. shRNA plasmids were used to suppress LMO2 in CAFs. LMO2 mRNA and protein level of both WPMY-1 and CAFs were evaluated by real-time fluorescent quantitative polymerase chain reaction (RTFQ-PCR) and Western blot. Then, PC-3 cells were co-cultured with different prostate stromal cells and the in vitro proliferation and invasion of PC-3 were measured by CCK-8 and matrigel invasion assays respectively. Results:When co-cultured with LMO2-overexpressed prostate stromal cells, both proliferation and in-vasion of PC-3 were improved. However, when co-cultured with CAFs which have inhibited expression of LMO2, the proliferation and invasion of PC-3 were reduced. The protein array proifling found that both interleukin-11 (IL-11) and ifbroblast growth factor-9 (FGF-9) were enhanced extensively in the supernatant collected from LMO2-overexpressed WPMY-1 cells. Conclusion:The expression of LMO2 in prostate stromal cells could be responsible for development of prostate cancer. Paracrine of cytokines, such as IL-11 and FGF-9, from LMO2-overexpressed stromal cells had effects on the proliferation and invasion of prostate cancer cells.

Objective The study was to find the application effect of high quality nursing service in perioperative treatment of patients with coronary heart disease.Methods Eighty patients were randomly divided into the comfort nursing group and the routine nursing group with 40 patients in each group.The comfort nursing group was given high quality nursing service,and the routine nursing group received the conventional care.The effect in two groups was compared after rehabilitation.The statistical software of SPSS 17.0 was used to analyze the data.Results The degree of anxiety was lower,the number of patients with complication was less,the degree of satisfaction was higher and the time in hospital was less in the comfort nursing group.Conclusions The comfortable nursing care should be used in peri-operative treatment of coronary heart disease,which can make patients keep the best mental state to accept and cooperate with the surgical treatment.It also can improve the care quality and patients' satisfaction degree and shorten the hospital stay.

AIM:To explore the changes of plasma levels of soluble vascular endothelial growth factor receptor 2 ( sVEGFR2) and superoxide dismutase ( SOD) in hypertensive patients and hypertensive diabetic patients.METHODS:In this cross-sectional study, 88 cases were enrolled, which were divided into hypertensive group (n=31), hypertensive diabetic group ( n=31 ) and control group ( n=26 ) .Blood pressure was obtained from each participant with mercury sphygmomanometer.The levels of sVEGFR2 and SOD were measured by ELISA.Meanwhile, the levels of plasma glucose, glycosylated hemoglobin A1c ( GHbA1c) and lipid profile were detected.RESULTS:The levels of total cholesterol ( TC) and body mass index (BMI) were significantly higher in hypertensive group than those in control group (P<0.05).The levels of TC, low-density lipoprotein cholesterol ( LDL-C) , triglyceride ( TG) , BMI, waist circumference were significantly higher in hypertensive diabetic group than those in control group (P<0.05).The plasma levels of sVEGFR2 and SOD in both hypertensive diabetic group and hypertensive group were significantly decreased compared with control group ( P<0.05), while the mean plasma levels of sVEGFR2 and SOD in hypertensive diabetic group were significantly decreased compared to the hypertensive group ( P<0.05 ) .A significantly positive correlation between sVEGFR2 and SOD in the whole study population (P<0.05) was observed.CONCLUSION: The plasma level of sVEGFR2 is decreased in both hypertensive and hypertensive diabetic patients, and more significantly decreased in hypertensive diabetic patients.De-creased SOD level may be associated with to the reduction of sVEGFR2.

Objective To evaluate the clinical efficacy of a stroke unit combined with community health services for treating stroke survivors. Methods A total of 120 stroke patients were randomly divided into a " stroke unit combined with community medicine" group ( combined group) , a stroke unit group and a general treatment group. Patients in the former 2 groups were treated in a hospital stroke unit during their hospitali-zation. The general treatment group was given conventional medical treatment. After discharge, the combined group continued to receive regular rehabilitation therapy and guidance in the form of community medical services, while the stroke unit group received follow-up only. Assessment was by means of Fugl-Meyer scores, the Barthel index and self-rating on a depression scale ( SDS). The patients were assessed at admission, on discharge and 3 months after discharge. Results There were no significant differences in average limb motor function, ability in the activities of daily living ( ADL) or depressive mood among the 3 groups on admission, but at discharge, limb motor function and ADL ability in the combined group and stroke unit groups were significantly superior to those in the general therapy group. Limb motor function and ADL ability in the combined and stroke unit groups had improved further 3 months after discharge, with more significant improvements in the combined group. No significant change in depression was observed in any group at discharge, but average depression scores in the combined and stroke unit groups improved significantly in the 3 months after discharge, and there was a statistically significant difference between the combined group and the general group. Conclusion Supplementing the work of a stroke unit with community health services significantly improves stroke patients' recovery of limb motor function and ADL ability.

Objective To clone and identify the heat shock factors(HSFs)of Schistosoma japonicum and analyze its molec?ular structure and alternative splicing pattern. Methods The New Zealand rabbits were infected with the cercariae of Schistoso?ma japonicum and were killed and dissected 42 days post?infection,and the adult worms of S. japonicum and the livers of the rabbits were harvested. Then,the total RNA was extracted by using Trizol reagent. The Sj?hsf open reading frame(ORF)and the alternative splicing fragments were amplified by RT?PCR from the female,male and egg samples,then cloned and verified by enzyme digestion and sequencing. DNAMAN 8.0,InterPro,Mega 6 combined with the Internet databases were utilized to clarify the gene structure,functional domains,alternative splicing pattern,and the homology and phylogenetic tree of HSFs. Re?sults Sj?hsf ORF and the alternative splicing fragments were amplified from the female,male and egg samples of S. japonicum by RT?PCR. After cloning,the positive recombinant plasmids pBSjHSFf?F,pBSjHSFf?M,pBSjHSFf?E containing Sj?hsf ORF, pBSjHSFs?F,pBSjHSFs?M,pBSjHSFs?E with Sj?hsf alternative splicing fragments were identified by enzyme digestion and se?quencing. Three alternative splicing Sj?hsf isoforms were observed through sequence analysis:Sj?hsf?isoform1(2 050 bp),Sj?hsf ?isoform2(2 086 bp)and Sj?hsf?isoform3(2 111 bp);the GenBank accession numbers were KU954546,KX119143 and KX119144,respectively. All the three isoforms located in the same Contig SJC_S000780 of S. japonicum genome and all ex?pressed at female,male and egg stages,but Sj?hsf?isoform1 with a high?level expression. Sj?HSF?isoform1(671 aa)and Sj?HSF?isoform2(683 aa)had DBD(DNA binding domain),HR?A/B and HR?C domains,while Sj?HSF?isoform3(282 aa)stopped in advance without HR?C domain. Phylogenetic tree analysis of HSFs illustrated that Sj?HSFs belonged to HSF1 family,with a close phylogenetic relationship to Sm?HSFs. Conclusions There are three alternative splicing isoforms of Sj?HSF existing in the female,male and egg stages of S. japonicum,but Sj?HSF?isoform1 expresses in a high?level. This study lays the foundation for further study on molecular mechanisms of Sj?HSFs in regulating the heat shock response system.

OBJECTIVETo delineate the clinical and genetic characteristics of patients with Allan-Herndon-Dudley syndrome (AHDS).

METHODSGenetic testing was carried out by next generation sequencing on 117 patients featuring intellectual disability and developmental delay. Clinical information including clinical manifestation, brain magnetic resonance imaging(MRI）, thyroid hormone levels, and electrocardiogram was collected for those with SLC16A2 mutations.

RESULTSFive male patients with SLC16A2 gene mutations were identified, including 2 affected brothers and 3 sporadic cases. The ages of the patients ranged from 8 months to 8 years. All patients presented with severe intellectual disability and developmental delay including poor head control, inability to sit independently, no speech, and poor response to external stimuli. All patients presented with hypotonia, dystonia, and positive pyramidal signs. Three patients had sinus tachycardia. All patients had abnormal thyroid hormone levels with elevated free triiodothyronine (FT), decreased free tetraiodothyronine(FT), and normal thyroid stimulating hormone (TSH). Brain MRI on 3 patients showed delayed myelination. Among the 3 sporadic patients, 2 carried de novo mutations including c.61G to T(p.E21X) and c.695_699delATGGT(p.N232SfsX7), respectively, 1 carried a c.42delC(p.W15GfsX69)mutation, which was inherited from his heterozygous mother. A nonsense mutation (c.916C to T, p.Q306X) was discovered in the two brothers, for which their mother was heterozygous.

CONCLUSIONAHDS is characterized by severe psychomotor developmental delay as well as congenital hypotonia, dystonia and positive pyramidal signs. Affected males may present with distinctive thyroid hormone abnormalities including increased FT and low FT accompanied by normal TSH. Delayed meylination of white matter is common. It is an X-linked mental retardation caused by SLC16A2 gene mutations.

Previous studies proposed that the synergistic effect of fibroblast growth factor-21 (FGF-21) and insulin may be due to the improvement of insulin sensitivity by FGF-21. However, there is no experimental evidence to support this. This study was designed to elucidate the mechanism of synergistic effect of FGF-21 and insulin in the regulation of glucose metabolism. The synergistic effect of FGF-21 and insulin on regulating glucose metabolism was demonstrated by investigating the glucose absorption rate by insulin resistance HepG2 cell model and the blood glucose chances in type 2 diabetic db/db mice after treatments with different concentrations of FGF-21 or/and insulin; The synergistic metabolism was revealed through detecting GLUT1 and GLUT4 transcription levels in the liver by real-time PCR method. The experimental results showed that FGF-21 and insulin have a synergistic effect on the regulation of glucose metabolism. The results of real-time PCR showed that the effective dose of FGF-21 could up-regulate the transcription level of GLUT1 in a dose-dependent manner, but had no effect on the transcription level of GLUT4. Insulin (4 u) alone could up-regulate the transcription level of GLUT4, yet had no effect on that of GLUT1. Ineffective dose 0.1 mg kg(-1) FGF-21 alone could not change the transcription level of GLUT1 or GLUT4. However, when the ineffective dose 0.1 mg x kg(-1) FGF-21 was used in combination with insulin (4 u) significantly increased the transcription levels of both GLUT1 and GLUT4, the transcription level of GLUT1 was similar to that treated with 5 time concentration of FGF-21 alone; the transcription level of GLUT4 is higher than that treated with insulin (4 u) alone. In summary, in the presence of FGF-21, insulin increases the sensitivity of FGF-21 through enhancing GLUT1 transcription. Vice versa, FGF-21 increases the sensitivity of insulin by stimulating GLUT4 transcription in the presence of insulin. FGF-21 and insulin exert a synergistic effect on glucose metabolism through mutual sensitization.

Objective: To study the mutational characteristics of KCNT1 and its clinical features in children with early-onset epileptic encephalopathy. Methods: Retrospective analysis of clinical data of 175 children with early onset epilepsy from the Department of Pediatrics at Peking University First Hospital from January 2012 to December 2017. Gene-based analysis was performed on children with targeted capture second-generation sequencing and the source of mutations was verified by PCR-Sanger. The clinical features of children with KCNT1 mutation were summarized. Results: In 175 infants with early-onset epileptic encephalopathy, 6 children were found to have KCNT1 mutations, all of which were new mutations with an overall mutation rate of 3.4% (6/175). All the mutations were missense mutations. The age of onset was from 2 days to 32 days. Five children were diagnosed with epilepsy of infancy with migrating focal seizure, one case was diagnosed with epilepsy, focal seizures, focal seizures with generalization. A total of 6 children were treated with multi-antiepileptic drugs. The disease in 4 patients were partially controlled, while in 2 patients, the disease was not significantly alleviated. One patient died of "severe pneumonia" at one year and 4 months of age. Then, four cases were treated with quinidine. The seizure frequency had no change in 3 cases, the frequency decreased and then relapsed in 1 case. The case once ketogenic diet and failed. Ketogenic diet treatment was applied to 5 cases, no significant effect was achieved. All the 6 patients had severe developmental delay. They could not sit alone, follow the light and objects and had no language. Conclusions The mutation of KCNT1 gene is mainly de novo. The onset of the disease was early, and mostly occurs in neonate and early infancy. The main seizure type was epilepsy of infancy with migrating focal seizure. Patients usually had severe psychomotor developmental delay. Antiepileptic drugs are ineffective. The efficacy of quinidine was not significant. Though, it still need studies on a large sample.