Peptides are increasingly important resources for biological and therapeutic development, however, their intrinsic susceptibility to proteolytic degradation represents a big hurdle. As a natural agonist for GLP-1R, glucagon-like peptide 1 (GLP-1) is of significant clinical interest for the treatment of type-2 diabetes mellitus, but its in vivo instability and short half-life have largely prevented its therapeutic application. Here, we describe the rational design of a series of α/sulfono-γ-AA peptide hybrid analogues of GLP-1 as the GLP-1R agonists. Certain GLP-1 hybrid analogues exhibited enhanced stability (t _1/2 > 14 days) compared to t _1/2 (<1 day) of GLP-1 in the blood plasma and in vivo. These newly developed peptide hybrids may be viable alternative of semaglutide for type-2 diabetes treatment. Additionally, our findings suggest that sulfono-γ-AA residues could be adopted to substitute canonical amino acids residues to improve the pharmacological activity of peptide-based drugs.

The responsive patterns of phytochrome gene family members to photoperiod and abiotic stresses were comparatively analyzed and the favorable natural variation sites of these genes were identified. This would help understand the mechanism of phytochrome gene family in photoperiod-regulated growth and development and abiotic stress response. In addition, it may facilitate the molecular marker assisted selection of key traits in foxtail millet. In this study, we used RT-PCR to clone three phytochrome genes SiPHYA, SiPHYB and SiPHYC from ultra-late maturity millet landrace variety 'Maosu'. After primary bioinformatics analysis, we studied the photoperiod control mode and the characteristics of these genes in responding to five abiotic stresses including polyethylene glycol (PEG)-simulated drought, natural drought, abscisic acid (ABA), high temperature and NaCl by fluorescence quantitative PCR. Finally, we detected the mutation sites of the three genes among 160 foxtail millet materials and performed haplotype analysis to determine the genes' functional effect. We found that the cloned cDNA sequences of gene SiPHYA, SiPHYB and SiPHYC were 3 981, 3 953 and 3 764 bp respectively, which contained complete coding regions. Gene SiPHYB and SiPHYC showed closer evolutionary relationship. Photoperiod regulated all of the three genes, but showed more profound effects on diurnal expression pattern of SiPHYB, SiPHYC than that of SiPHYA. Under short-day, when near heading, the expression levels of SiPHYA and SiPHYB were significantly lower than that under long-day, indicating their roles in suppressing heading of foxtail millet under long-day. SiPHYB and SiPHYC were responsive to PEG-simulated drought, natural drought, ABA and high temperature stresses together. SiPHYA and SiPHYB responded differently to salt stress, whereas SiPHYC did not respond to salt stress. Re-sequencing of 160 foxtail millet materials revealed that SiPHYB was highly conservative. Two missense mutations of SiPHYA, such as single nucleotide polymorphism (SNP) 7 034 522^C→T and SNP7 036 657^G→C, led to delaying heading and increasing plant height. One missense mutation of SiPHYC, such as SNP5 414 823^G→T, led to shortening heading under short-day and delaying heading under long-day, as well as increasing plant height and panicle length regardless of photo-thermal conditions. Photoperiod showed different regulatory effects on SiPHYA, SiPHYB and SiPHYC. SiPHYB and SiPHYC jointly responded to various abiotic stresses except for the salt stress. Compared with the reference genotype, mutation genotypes of SiPHYA and SiPHYC delayed heading and increased plant height and panicle length.
Gene Expression Regulation, Plant ; Photoperiod ; Phytochrome/metabolism* ; Plant Proteins/metabolism* ; Setaria Plant/metabolism* ; Stress, Physiological/genetics*

OBJECTIVE： To construct prediction model for initial dose of levothyroxine （L-T4） in differentiated thyroid cancer （DTC） patients after surgery. METHODS： A total of 100 DTC patients underwent surgery were selected from thyroid and breast surgery department in Nanjing Drum Tower Hospital. General information of patients such as gender， age， height， body mass， body mass index （BMI） and regular follow-up information after discharge were collected. Related data of thyroid function and adjusted dose of L-T4 were recorded. Single factor variance analysis and t-test were used to analyze the predictors that had significant correlation with the initial dose of L-T4. The prediction model of L-T4 initial dose was established by linear regression analysis， and was verified by prospective experiments. RESULTS： The initial dose of L-T4 in DTC post-surgery patients were significantly correlated with age （P＝0.01，F＝3.993）， body weight （P＜0.001，F＝6.910） and BMI （P＜0.001，F＝7.698）. Linear regression analysis showed that prediction model of initial dose of L-T4 was L-T4（μg/kg）＝2.971-0.033×BMI-0.005×age. DTC post-operative patients were given L-T4 empirically， and only 16％ （16/100） of the patients met the criteriaat the first follow-up  of thyroicl function. In the validation test， L-T4 was given at the initial dose calculated by the prediction model， and 63.7％      （44/69） of the patients reached the standard at the first follow-up. CONCLUSIONS： The established prediction model of L-T4 initial dose after DTC surgery has a certain practicality.

Objective To establish a porcine model of liver failure after different percent hepatectomy.Methods The porcine models of liver failure 75%,85%,95% hepatectomy were developed and the living conditions and survival time were recorded.The blood samples of pre-surgery,post-hepatectomy d1,d3,d5 and post-hepatectomy 1 week,2 weeks,and 3 weeks were collected for hepatic function analysis.Histological examination of liver tissues was performed using HE staining.Liver injury histology was interpreted and scored in the terminal samples.Results The average survival time of pigs with post-hepatectomy liver failure after 75%,85%,95% hepatectomy was 19.0±5.6 days,17.3±5.5 days,1.3±1.5 days,respectively.Their pathological scores were 5.67±0.52,8.17±0.82 and 8.50±0.71,respectively.With the increase of percent hepatic resection,the incidence of hepatic failure was increasing.ALT,AST,ALP,LDH and TBA were dramatically increased in the pigs after 85% hepatectomy.Conclusions The pig model of acute liver failure by 85% hepatectomy is successfully established,which can cause typical acute liver failure in Bama miniature pigs.

Objective To study the synergetic effect and possible mechanism of transplanting mesenchymal stem cells (MSCs) in combination with interleukin-1 receptor antagonist (IL-1Ra) on acute liver failure (ALF).Methods MSCs transplantation combined with IL-1Ra was used for a swine model of ALF induced by 85％ total hepatectomy.The living conditions,blood samples and survival time were recorded or collected for analysis of hepatic function.Liver injury histology was analyzed.Hepatic cell regeneration and apoptosis were studied by immunohistochemistry staining of Ki67 and TUNELassays respectively.The expression levels of AKT and NF-κB were analyzed by Western blotting.Results The difference on the survival time between the model group and combined therapy group was statistically significant (P ＜ 0.05).Combined therapy displayed improvement not only in the serum biochemical conditions but also in the serum inflammatory cytokines.Furthermore,the observed hepatic histopathological score was significantly less compared to model group.In addition,the combined therapy group significantly inhibited the liver cell apoptosis and increased hepatic cell regeneration.Finally,a significant increase in AKT expression and decrease of NF-κB expression (P ＜ 0.05) were observed,which was consistent with their important roles in liver regeneration.Conclusion The combined therapy displayed a synergistic effect on liver regeneration,by promoting restoration and reconstruction of ALF,through regulation of inflammation and apoptosis signaling network.

Objective To explore the related risk factors of central lymph node metastasis ( CLNM) of papillary thyroid microcarcinoma ( PTMC) and the application value of central lymph node dissection ( CLND) . Methods Totally 251 patients who were confirmed as PTMC by pathological biopsy were selected as the research object, and they were divided into the metastasis group (n=42) and the non-metastasis group (n=209) according to the status of lymph node metastasis. The general datum of the two groups were retrospectively analyzed, and the indexes with significant differences after univariate analysis were analyzed by non-conditional Logistic regression analysis. Treated the patients with lymph node metastasis through CLND, and the occurrence rate and recurrence rate of related complications were recorded during the 1 to 3 years of follow-up. Results The comparison of the location of the tumor, complicated adenoma, Hashimoto’ s thyroiditis, nodular goiter, and hyperthyroidism between the two groups was not statistically significant (P>0. 05). In the metastasis group, the rate of patients aged less than 45 years old and the percentage of male patients were both significantly higher than the non-metastasis group, and the difference was of statistical significance (P<0. 05). There were 25 cases (59. 5%) whose tumor diameter were 5~10 mm, 12 cases (28. 6%) with multiple lesions, and 22 cases (52. 4%) breaking through the envelope in the metastasis group, which were significantly higher than 70 cases (33. 5%), 23 cases (11. 0%) and 67 cases (32. 1%) in the non-metastasis group, and the differences were statistically significant (P<0. 05). In the metastasis group after operation, there was 1 cases (2. 3%) of hoarseness, 6 cases (14. 3%) of local numbness, 1 case (2. 3%) of nerve invasion, and 2 cases (4. 8%) of vascular invasion, and there was no choking cough report. During the period of 1 to 3 years of follow-up, there were only 2 cases of recurrence and the recurrence rate was 4. 8%. Conclusion Central lymph node metastasis mostly occurs in the male patients with PTMC who are less than 45 years old. And the main characteristics are diameter of tumor over 5 mm, multifocal lesions, with envelope penetration symptom and so on. In clinic, patients who meet the above-mentioned condition should be taken as the primary concern and given related prevention measures to reduce the occurrence risk of central lymph node metastasis. To treat patients with papillary thyroid microcarcinoma through central lymph node dissection can receive good curative effect and effectively improve the prog-nosis quality, and it is worthy of clinical promotion.

Objective To explore the most effective route of mesenchymal stem cell transplantation (MSCs) in D-galactosamine (D-gal) induced porcine model with acute liver failure (ALF) and the potential mechanism.Methods BA-MA mini-pigs with D-gal-induced ALF were transplanted with porcine mesenchymal stem cells (MSCs) through four routes:intraportal injection (InP),peripheral intravenous injection (PV),hepatic intra-arterial injection (AH) and intrahepatic injection (IH).The survival time was recorded.The blood samples before and after MSC transplantation were collected for detecting liver function.Liver histology was interpreted and scored.Hepatic apoptosis and regeneration were detected by Terminal Deoxynucleotidyl Transferase dUTP Nick End Labeling (TUNEL) assay and Ki-67 assay.The protein expression of cleaved caspase-3,survivin,AKT and ERK were analyzed by Western blot.Results The average survival time in each group was (10.7 ±1.6) days (InP),(6.0 ±0.9) days (AH),(4.7 ±1.4) days (PV),(4.3 ± 0.8) days (IH) when compared with D-gal group [(3.8 ± 0.8) days].The histopathological scores revealed a significantly decrease in InP group (3.17 ± 1.04,P ＜0.05) and AH group (8.17 ± 0.76,P ＜ 0.05) when compared with that in D-gal group (11.50 ± 1.32).The apoptosis rate in InP group (25.0 ± 3.4％,P ＜ 0.05) and AH group (40.5 ± 1.0％,P ＜ 0.05) was lower than that in D-gal group (70.6 ± 8.5％).The expression of active caspase-3 was inhibited,while the expression of survivin,AKT and ERK was elevated in InP group.Conclusions The intraportal injection was superior to other pathways for MSCs transplantation.Intraportal MSC transplantation could improve liver function,inhibit cell apoptosis,promote cell proliferation and prolong the survival in porcine ALF model.

Surgery is so far the most widely used and effective treatment of neoplastic diseases.However,residual tumour cells during surgery remain a major trigger of cancer recurrence and matastasis.Although intraoperative rapid pathological R0 resection can be achieved based on preoperative imageological examination,but for small satellite lesions and the naked eye can not find the error quickly and so often cause pathological presence of residual tumour cells.Thus,quick and accurate identification of residual cancer cells is crucial for prognosis of cancer patients.Indocyanine green (ICG) is a new type of fluorochrome that can stain tumours under the near-infrared fluorescence during surgery,the paper will be reviewed latest developments in the reagent for fluorescence in tumours.

Objective To investigate the congenital heart disease (CHD) prevalence of students in Liangshan prefecture in Sichuan province,and to explore the risk factors of CHD.Methods Stratified cluster sampling was conducted in collecting demographic data.Heart auscultation was used to screening the CHD,and colour Doppler flow imaging was used to diagnose the CHD.Results A total of 10 021 student were collected in this study.About 19.74％ of total cases had cardiac murmur.Sixty-eight students were diagnosed as CHD through colour Doppler flow imaging,accounting for 0.68％.In addition,the top three ranks of CHD were atrial septal defect,patent ductus arteriosus,interventricular septal defect.Our study also found that per capita income of CHD families was lower than non-CHD families,meanwhile,the CHD prevalence of Yi nationality was higher than other nationalities.There was no statistical significance in age,gender,height,weight,body mass index,head circumference,waist circumference between different types of CHD.Conclusions The CHD prevalence of students in Liangshan prefecture has been in a high level in China; therefore we need pay more attention on screening for the CHD.

To study the MEN1 gene mutations in a multiple endocrine neoplasia type 1 ( MEN 1 ) family,and determine the possible mechanism of disease induced by the mutations.Genomic DNA was isolated from peripheral blood leukocytes and the MEN1-related tumor tissues of the patient and the family members,then the coding exons and exon/intron boundaries of the MEN1 gene were amplified by polymerase chain reaction (PCR) and sequenced.Subclone sequencing was performed to identify the heterozygosity.Further immunohistochemistry was performed to observe menin protein expression in the tumor tissues.We identified a heterozygous deletion mutation of intron 9 ( IVS9+ 1_11 delGTGAGGGACAG) in the proband and two family menbers.We also demonstrated for the first time that the expression of menin protein is absent in the parathyroid adenoma tissue.The heterozygous mutation in the initial of intron 9,IVS9+ 1_11 delGTGAGGGACAG is a new type of MEN1 gene mutations in China.This mutation may produce an aberrant splicing of MEN1 mRNA,generating easily degradation and loss of expression of menin protein and resulting eventually in the disease.