Abstract: This study is to improve the affinity of scFv-AK404R against VEGFR2. The secondary mutational library was constructed by hydrophilic shuffling in CDR3 region of the heavy chain. VEGFR2-specific screening was performed by phage display technology and the protein of mutants was expressed in periplasm of E.coli HB2151 and purified by affinity chromatography. The affinity constant of scFvs was measured by competitive ELISA, and the structure of scFvs was analyzed by bioinformatics. The result showed that a library with 6.4x10(5) scFv members was established by electro-transformation. Two mutated clones with high absorbance value were isolated after screening. After purification by affinity chromatography, electrophoretically pure scFv proteins were obtained. The competitive ELISA showed that the affinities of WZ01 and WZ02 were three times higher than that of the parental AK404R, and bioinformatics analysis showed that the enlarged contact surface and fitted closely with KDR3 surface may be the reasons for improved affinity. These results suggest that introducing hydrophilic amino acids to the heavy chain CDR3 region is an effective approach to improve the affinity of scFv.
Amino Acid Sequence ; Antibody Affinity ; Chromatography, Affinity ; Computational Biology ; Enzyme-Linked Immunosorbent Assay ; Escherichia coli ; metabolism ; Hydrophobic and Hydrophilic Interactions ; Peptide Library ; Single-Chain Antibodies ; genetics ; immunology ; isolation & purification ; Vascular Endothelial Growth Factor Receptor-2 ; genetics ; immunology ; metabolism

OBJECTIVETo investigate the selectivity and individualization of transpedicular balloon kyphoplasty for aged osteoporotic spinal fracture.

METHODSTwenty-two consecutive procedures were performed in 17 aged patients with osteoporotic spinal compression fractures from April 2002 to June 2004. The signal changes in different sequences were confirmed by magnetic resonance imaging before the procedures. This operation involved the percutaneous insertion of two inflatable bone tamps into a fractured vertebral body transpedicularly under fluoroscopic guidance. Every patient was treated individually, according to the results of radiography and CT scan before operation. Preoperative and postoperative complications, visual analogue scale, and radiographic findings such as vertebral height and Cobb angle were recorded and analyzed.

RESULTSAll patients tolerated the procedure well with immediate relief of their back pain in 24 hours. There was no leakage of cement into the epidura. The mean loss percent of the anterior and middle vertebral heights were (35.32 +/- 13.15)% and (27.53 +/- 12.61)% before operation, and (14.21 +/- 12.43)% and (16.2 +/- 7.5)% after operation. The height restoration of vertebra was confirmed by X-ray after the procedure (P < 0.01). The mean kyphosis was improved from (25.3 +/- 4.2) degrees to (8.6 +/- 5.1) degrees. No complications occurred. No patient had nerve injury. The patients were allowed to walk next day after the procedure.

CONCLUSIONThe selectivity and individualization of transpedicular balloon kyphoplasty for aged osteoporotic spinal fractures has satisfactory short-term clinical efficacy. It is also an effective way to prevent complications.

Aged ; Aged, 80 and over ; Female ; Fracture Fixation, Internal ; methods ; Fractures, Spontaneous ; etiology ; surgery ; Humans ; Kyphosis ; etiology ; surgery ; Male ; Middle Aged ; Minimally Invasive Surgical Procedures ; methods ; Osteoporosis ; complications ; Osteoporosis, Postmenopausal ; complications ; Spinal Fractures ; surgery

OBJECTIVETo compare the efficacy and safety of PEG-IFNalpha-2b (Peg-Intron) with IFNalpha-2b (Intron A) in treating HBeAg positive chronic hepatitis B patients.

METHODSTwo hundred thirty chronic hepatitis B (CHB) patients eligible to the following criteria were enrolled into this study: HBsAg and HBeAg(Abbott kit) positive for at least 6 months, serum HBV DNA > or =10(5) copies/ml (real time PCR, LLQ <10(3) copies/ml) and ALT > or =2 x ULN. After 1:1 randomization, the patients received PegIntron (group A: 1.0 microg/kg body weight, SC, once a week) or Intron A (group B: 3 MIU SC, three times a week) for 24 weeks, and followed up for 24 weeks.

RESULTS(1) In groups A and B, respectively, 80.87% and 83.48% were males; their median ages were 31.0 and 32.0 years old; their median body weights were 65.6 and 65.5 kg; mean serum HBV DNA loads were 8.06 log10 and 7.99 log10; their mean ALT values were 4.17 x ULN and 3.77 x ULN. All of the above parameters between the two groups had no statistically significance differences. (2) At the end of treatment and after follow-up, compared to the Intron A group, the PegIntron group showed better response (including complete and partial response rate, HBV DNA undetectable rate, HBeAg seroconversion rate), but the differences of all of them had no statistical significance. The rate of HBeAg loss was higher in patients receiving PegIntron after follow-up (P = 0.0424). (Table 2) (3) PegIntron and Intron A reduced serum HBV DAN persistently during the therapy. Mean reduction at the end of the treatment was much higher in the PegIntron group than in the Intron group (2.22 log10 copies/ml vs 1.66 log10 copies/ml, P = 0.0283). (4) The overall incidence of adverse events (AEs) in the PegIntron group was similar to that of the Intron A group (94.78% vs 95.65%). The AEs associated with PegIntron administration were similar in nature to those with Interon A, such as influenza-like symptoms, fever, fatigue, headache, nausea, etc and the differences of their incidences had no statistical significance.

CONCLUSIONSThe efficacy and safety of PEG-IFNalpha-2b treatment for CHB patients seems to be better than that of IFNalpha-2b; however, further studies are needed to confirm it.

Adolescent ; Adult ; Aged ; Antiviral Agents ; adverse effects ; therapeutic use ; Female ; Hepatitis B e Antigens ; blood ; Hepatitis B, Chronic ; drug therapy ; immunology ; Humans ; Interferon-alpha ; adverse effects ; therapeutic use ; Male ; Middle Aged ; Polyethylene Glycols ; Recombinant Proteins

OBJECTIVETo explore how to determine painful vertebrae treated by kyphoplasty in multiple-level osteoporotic vertebral compression fractures and clinical outcome.

METHODSFrom October 2002 to June 2005, 51 consecutive procedures with kyphoplasty were performed on 35 patients with multiple-level osteoporotic vertebral compression fractures. There were 51 painful vertebrae among 120 vertebral compression fractures. The painful vertebra was determined by the signal intensity changes in MR images, combined with radiography and local percussion pain before operation. Only painful vertebrae were treated by kyphoplasty. Preoperative, postoperative and final follow-up visual analog scale (VAS) and radiographic findings such as vertebral height and Cobb angle were recorded and analyzed.

RESULTSAll patients tolerated the procedure well with immediate relief of their back pain after kyphoplasty and they can walk at 1-3 days after the procedure. There were 3 vertebrae (3/51) occurred asymptomatic extravasation of cement. 31 cases were followed up for mean 16.2 months (range 6-44 months). VAS reduced from preoperative 8.7 to final follow-up 2.1 (P <0. 01). At final follow-up the vertebral height had a recovery rate of 59.17%, and the mean Cobb angle was improved 10.1 degrees. There was a significant improvement between preoperative and final follow-up values (P < 0.01).

CONCLUSIONSThe painful vertebra can be determined by signal intensity changes in MR series images in multiple-level osteoporotic vertebral compression fractures. Selecting painful vertebrae to be treated by kyphoplasty can make patients with multiple-level VCFs gain an excellent result.

Adult ; Aged ; Aged, 80 and over ; Female ; Follow-Up Studies ; Fractures, Compression ; diagnosis ; etiology ; surgery ; Humans ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Osteoporosis ; complications ; Prospective Studies ; Spinal Fractures ; diagnosis ; etiology ; surgery ; Treatment Outcome ; Vertebroplasty ; methods

BACKGROUND AND OBJECTIVEInvasion and metastasis are the most common causes of mortality for patients with colorectal neoplasms, and blocking invasion and metastasis in a timely fashion has become a hot research focus. We investigated the expression of the messenger RNA of Syndecan-1 and HPA-1 in colorectal cancer, and their correlation with invasion and metastasis.

METHODSReal-time fluorescent quantitative polymerase chain reaction (PCR) was used to detect the expression of Syndecan-1 and HPA-1 in specimens from 49 patients with colorectal cancer, 49 paired adjacent colorectal neoplasms (2 cm from the carcinoma), and 49 surgical margins of paired normal colorectal mucosa tissue (5 cm from the carcinoma), to analyze their correlation with clinicopathologic characteristics of colorectal neoplasm.

RESULTSThe expression of HPA-1 mRNA was significantly higher in colorectal cancer (40.56 +/- 11.75) than that in the paired adjacent colorectal neoplasms (18.28 +/- 11.33) and normal colorectal mucosa tissue (10.80 +/- 10.20) (all P < 0.001). The expression of HPA-1 mRNA was significantly higher in paired adjacent colorectal neoplasms than that in normal colorectal mucosa (P < 0.05). The expression of Syndecan-1 mRNA was significantly higher in normal colorectal mucosa (61.21 +/- 12.96) than in the paired adjacent mucosa (14.35 +/- 11.06) or colorectal cancer (10.12 +/- 8.58) (all P < 0.001). The expression of Syndecan-1 mRNA was significantly higher in the paired adjacent mucosa than that in colorectal cancer (P < 0.05). The decreased expression of Syndecan-1 mRNA and the increased expression of HPA-1 were closely associated with the degree of differentiation, the depth of infiltration, lymph node metastasis, vessel metastasis, and TNM staging of colorectal cancer (all P < 0.05). Spearman rank correlation analysis demonstrated a significant correlation between Syndecan-1 and HPA-1(r = -0.405, P < 0.05).

CONCLUSIONSThe expression of Syndecan-1 mRNA was significantly highest in normal colorectal mucosa and the expression of HPA-1 mRNA was significantly highest in colorectal cancer. At the same time, the decreased expression of Syndecan-1 mRNA and the increased expression of HPA-1 mRNA can promote the invasion and metastasis of colorectal cancer. The determination of Syndecan-1 and HPA-1 may be of value in the treatment as well as in the prognosis of patients with colorectal cancer.

Adenocarcinoma ; metabolism ; pathology ; Colorectal Neoplasms ; metabolism ; pathology ; Gene Expression Regulation, Neoplastic ; Glucuronidase ; genetics ; metabolism ; Humans ; Intestinal Mucosa ; metabolism ; pathology ; Lymphatic Metastasis ; Neoplasm Invasiveness ; Neoplasm Staging ; Prognosis ; RNA, Messenger ; metabolism ; Real-Time Polymerase Chain Reaction ; Syndecan-1 ; genetics ; metabolism

Objective:Continuous glucose monitoring (CGM) technology is used to compare the advantages of insulin degludec (IDeg) as a basal insulin regimen compared with insulin glargine (IGlar) in the treatment of adult type 1 diabetes mellitus.Methods:30 adult patients with T1DM admitted to Heji Hospital Affiliated to Changzhi Medical College from September 2019 to December 2020 were screened. According to the random number table method, the patients were randomly divided into two groups (insulin degludec group and insulin glargine group) at a ratio of 1∶1, respectively treated with IDeg, IGlar and aspartate insulin for 12 weeks. The main outcome measures were the coefficient of variation of blood glucose (CV), mean amplitude of glycemic excursions (MAGE), time in range (TIR), time above range (TAR) and time below range (TBR). The secondary outcome measures were mean blood glucose (MBG), standard deviation of blood glucose (SD), fasting blood glucose (FPG), 2 h postprandial blood glucose (2 h BG), hemoglobin A1c (HbA 1c), means of daily differences (MOOD), and the frequency of hypoglycemic events. Results:At 12 weeks of treatment, the HbA 1c, FPG, 2 h BG, MBG, SD, CV and MAGE of insulin degludec group were lower than those of insulin glargine group, with statistically significant difference (all P<0.05). The TIR in the insulin degludec group was significantly higher than that in the insulin glargine group [73(63, 75)% vs 43(28, 63)%, P<0.001], and the TAR was lower than that in the glycerine group [25(17, 23)% vs 35(33, 64)%, P=0.003]. From the curve spectrum of blood glucose level of the two groups, the stability of blood glucose in the insulin degludec group was better than that in the insulin glargine group. After 12 weeks of treatment, 8 cases (8/15) in insulin degludec group had HbA 1c<7.0%, and 4 cases (4/15) in insulin glargine group had HbA 1c<7.0%, without statistically significant difference ( P=0.264). There were 7 cases (7/15) in the insulin degludec group and 1 case (1/15) in the insulin glargine group who achieved high quality blood glucose control, with statistically significant difference ( P=0.035). At the 12th week of outpatient follow-up, the incidence of nocturnal hypoglycemic events in insulin degludec group was significantly lower than that in insulin glargine group (4/15 vs 11/15, P=0.027). Conclusions:Compared with insulin glargine, insulin degludec can achieve higher blood glucose compliance rate, lower blood glucose level and reduce blood glucose fluctuations in patients with type 1 diabetes.

Background and Objective:Invasion and metastasis are the most common causes of mortality for patients with colorectaI neoplasms,and blocking invasion and metastasis in a timely fashion has become a hot research focus.We investigated the expression of the messenger RNA (mRNA)of Syndecan-1 and HPA-1 in colorecta J cancer,and their correlation with invasion and metastasis.Methods:Real-time fluorescent quantitative reverse transcription polymerase chain reaction(RT-PCR)was used to detect the expression of Syndecan-1 and HPA-1 in specimens from 49 patients with colorectal cancer.49 paired adjacent colorectal neoplasms(2 cm from the carcinoma),and 49 surgical margins of paired normal colorectal mucosa tissue(5 cm from the carcinoma),to analyze their correlation with clinicopathologic characteristics of colorectal neoplasm.Results:The expression of HPA-1 mRNA was significantly higher in colorectal cancer (40.56±11.75)than that in the paired adjacent colorectal neoplasms(18.28±11.33)and normal colorectal mucosa tissue(10.80±1020)(all P＜0.001).The expression of HPA-1 mRNA was significantly higher in paired adjacent colorectal neoplasms than that in normal colorectal mucosa(P＜0.05).The expression of Syndecan-1 mRNA was significantly higher in normal colorectal mucosa(61.21±12.96)than in the paired adjacent mucosa (14.35±11.06)or colorectaI cancer(10.12±8.58)(all P＜0.001).The expression of Syndecan-1 mRNA was significantly higher in the paired adjacent mucosa than that in colorectal cancer(P＜0.05).The decreased expression of Syndecan-1 mRNA and the increased expression of HPA-1 were closely associated with the degree of differentiation,the depth of infiltration,lymph node metastasis,vessel metastasis,and TNM staging of colorectal cancer(all P＜0.05).Spearman rank correlation analysis demonstrated a significant correlation between Syndecan-1 and HPA-1(r=-0.405,P＜0.05).Conclusions:The expression of Syndecan-1 mRNA was significantly highest in normal colorectal mucosa and the expression of HPA-1 mRNA was significantly highest in colorectal cancer.At the same time,the decreased expression of Syndecan-1 mRNA and the increased expression of HPA-1 mRNA can promote the invasion and metastasis of colorectal cancer.The determination of Syndecan-1 and HPA-1 may be of value in the treatment as well as in the prognosis of patients with colorectal cancer.

BACKGROUNDChronic liver diseases are a major burden in China. Alanine aminotransferase (ALT) can be used as an indicator of hepatocyte damage. In this study, we determined the prevalence and etiologies of elevated ALT in an adult population in Jilin, China.

METHODSA total of 4072 individuals aged between 18 and 79 years were first interviewed, and then underwent ultrasonography and blood tests.

RESULTSThe prevalence of elevated ALT was 17.53%. The most noticeable risk factor for ALT elevation was non-alcoholic fatty liver disease (NAFLD) (accounting for 10.79%), metabolic syndrome (16.25%), or both (20.31%). The development of NAFLD occurred mostly in female peasants and small businessmen with increased income, age, fasting plasma glucose, body mass index, triglyceridemia, and low-density lipoprotein and decreased education level, high-density lipoprotein. Elevated ALT frequently occurred in low education level, male peasants and small businessmen with increased income, body mass index and triglyceride who had NAFLD and/or metabolic syndrome. However, elevated ALT with infection of hepatitis B or C virus was not associated with metabolic disorders, but rather with gender, occupation and increased age.

CONCLUSIONThe results from the current study demonstrate that elevated ALT is fairly high in the Northeast population (17.53%) and that the cause of its elevation is mostly due to NAFLD and metabolic syndrome.

Adolescent ; Adult ; Age Factors ; Aged ; Alanine Transaminase ; blood ; Fatty Liver ; blood ; epidemiology ; Female ; Hepatitis B ; blood ; epidemiology ; Hepatitis C ; blood ; epidemiology ; Humans ; Male ; Metabolic Syndrome ; blood ; Middle Aged ; Non-alcoholic Fatty Liver Disease ; Sex Factors ; Young Adult

OBJECTIVETo contrast single and double balloon-inflated kyphoplasty for vertebral compression fractures (VCFs) and evaluate its clinical efficacy.

METHODSFrom May 2000 to May 2004, 90 consecutive procedures were performed in 58 patients who suffered from painful vertebral compression fractures, transferring tumour and angioma. Ninety vertebrae were inflated while 62 as A group were double balloon and 28 as B group were single balloon, fracture reduction and bone cement augmentation. Preoperative and postoperative symptom levels, variables, complications were recorded and the vertebral height and Cobb angle were measured and analyzed.

RESULTSAll patients' pain was alleviated or disappeared without syndrome, and the vertebral height and Cobb angle of both groups were improved. The average recovery rate was 72.6% (22.9% approximately 100%), Cobb angle from 17.9 degrees (3.1 degrees approximately 31.6 degrees ) were corrected to 9.6 degrees (0.6 degrees approximately 28.2 degrees ), the average angle was 8.7 degrees (0.3 degrees approximately 27.2 degrees ), and the contrast between preoperative and postoperative showed obvious differences (P <0.001). The average recovery rate of A group was 77.6% (55.3% approximately 100%), B group was 64.3% (22.9% approximately 100%). The average postoperative Cobb angle of A group was 9.9 degrees (0.3 degrees approximately 27.2 degrees ), B group was 8.6 degrees (0.6 degrees approximately 19.8 degrees ) (P >0.05).

CONCLUSIONSAs a promising minimally invasive surgery, balloon kyphoplasty can provide early relief of pain and improve the function as well as spinal alignment in treatment of painful compression fracture owing to recovering the vertebral height and Cobb angle of the vertebral body. Single balloon-inflated kyphoplasty can improve VCFs as double balloon.

Adult ; Aged ; Aged, 80 and over ; Bone Cements ; therapeutic use ; Female ; Fractures, Compression ; complications ; surgery ; Humans ; Kyphosis ; etiology ; surgery ; Lumbar Vertebrae ; injuries ; surgery ; Male ; Middle Aged ; Minimally Invasive Surgical Procedures ; Orthopedic Procedures ; instrumentation ; methods ; Retrospective Studies ; Spinal Fractures ; complications ; surgery ; Thoracic Vertebrae ; injuries ; surgery

Diabetic kidney disease(DKD)is one of the most important complications of diabetes.In recent years,domestic and foreign studies have found that mammalian target protein of rapamycin(mTOR)related signaling pathway is a classic pathway involved in the regulation of autophagy,which can achieve the therapeutic effect of DKD by targeting the autophagy pathway,and plays a crucial role in the prevention and treatment of DKD.In this paper,we reviewed the mechanism of mTOR-related signaling pathway targeted autophagy in the prevention and treatment of DKD,in order to provide a new reference and basis for clinical prevention and treatment of DKD.