To rapidly select potent anti-VSTM1-v2 scFv (single-chain antibody fragment) by construction and screening of a humanized scFv library in which a murine VH-CDR3 library was grafted onto a human scFv framework. A murine VH-CDR3 library was amplified from anti-VSTM1-v2 murine cDNA and grafted on human scFv (VH3-VK1) framework. Anti-VSTM1-v2 scFv templates were selected and enriched through ribosome display, TA-cloned into expression vector, and transformed into BL21 (DE3) for soluble expression of target scFv. A total of 1000 clones were randomly picked. Positive ones were first identified using colony PCR, indirect ELISA, Western blotting and then verified with sequencing and dose response ELISA. At last an anti-VSTM1-v2 humanized scFv with good binding affinity (EC50 = 21.35 nmol x L(-1)) was selected from the humanized library of 10(12) members generated in this study. This scFv antibody might have potential applications. This study provides a new approach for rapid screening of humanized antibodies.

Objective To evaluate the efficiency of different interventional mode in treating primary hepatocellular carcinoma with decompensated liver cirrhosis.Methods 132 cases with hepatocellular carcinoma accompanied by decompensational liver cirrhosis were undertaken transcatheter arterial chemo-embolization(TACE,Group A,n=36),segmental transcatheter arterial chemo-embolization(S-TACE,Group B,n=54)and S-TACE + radiofrequency ablation(RFA) + percutaneous ethanol injection(PEI)(multi-interventional therapy,Group C,n=42).Univariate analysis and multivariate COX proportional hazard regression model were used to analyze the factors affecting the prognosis.Results All patients of A,B,C groups had been followed up for 3 months to 48 months,with median survival times of 4.1 months,9.4 months(P 50%) demonstrated as 8.3%,31.5%(P

Objective To investigate the prevalence and risk factors of metabolic syndrome (MS) in residents in Pudong New District of Shanghai. MethodsA total of 5 584 residents aged 20-80 years were randomly selected from Pudong New District of Shanghai through multistage sampling and interviewed from April to July of 2008. Metabolic syndrome was defined according to three diagnostic criteria for MS, issued by the modified National Cholesterol Education Program Adult Treatment Panel Ⅲ criteria ( NCEP-ATP Ⅲ ), International Diabetes Federation (IDF), and Chinese Diabetes Society (CDS). ResultsThe crude prevalences of MS in the adult population in Pudong New District were 18.2％ and 13.1％ standardized ( male 19. 1％, female 17.4％, the age-standardized 15.6％ and 13.2％ ) with CDS criterion, 31.8％ and 24.4％ standardized ( male 28.4％ ,female 35.1％ ,the agestandardized 22. 7％ and 25.0％ ) with NCEP-ATP Ⅲ criterion, and 21.7％ and 17.0％ standardized ( male 15.9％ ,female 26.7％, the age-standardized 13.8％ and 19.2％ ) with IDF criterion. The age-specific prevalence of MS increased according to three diagnostic criteria, and the age-adjusted prevalence was higher in males than females in junior age groups and higher in females than males in senior ones. Significant differences were present among region, education, marriage status, smoking, work intensity, recreation, and physical activity according to some diagnostic criteria. ConclusionsSubstantial proportions of adults in Pudong New District of Shanghai suffer from metabolic syndrome, and there exists a tendency for young people involved. MS has become a noteworthy public health problem. It suggests that community-integrated control strategy of MS should be made a priority.

As the specific endangered medicinal plant in Xinjiang, resources and distribution of Ferula sinkiangen-sis are important for biodiversity conservation and sustainable development of Chinese medicine resources. The spa-tial distribution and resources of F. sinkiangensis were researched based on low altitude remote sensing and sample investigation. The results showed that the optimum working time for F. sinkiangensis monitoring by low altitude remote sensing was the nearby 5 hills, which covered about 0.88 km2. It was suggested that the fence area should be expanded for protection. According to the results of low altitude remote sensing, the amount of F. sinkiangensis in yellow (diameter exceeding 0.3 m) was about 3 191. However, the sample investigation on amount of F. sinkiangensis in yellow (diameter exceeding 0.3 m) was about 2 752. The error between them was 14%. The monitoring time and range for F. sinkiangensis by low altitude remote sensing were also ensured. It was concluded that low altitude re-mote sensing had the advantage of quickly receiving distribution situation of F. sinkiangensis, which can effectively evaluate dynamic changes of F. sinkiangensis in Xinjiang.

Objective To study the CT and MR characteristic features of the respiratory epithelial adenomatoid hamartoma of olfactory clefts. Methods (1)The CT and MRI findings of 29 patients with histologically proved respiratory epithelial adenomatoid hamartoma in the olfactory clefts were retrospectively reviewed.All patients underwent CT and 8 of them underwent MRI. Location, CT and MRI features, and associated findings of the disease were reviewed;(2)The CT findings, olfactory clefts width, total nasal distance, and the ratio of OC to the total nasal distance of the case patients (29 cases) and the control patients (33 patients with sinusitis) were compared to investigate the correlation of the olfactory clefts distance and the incidence of respiratory epithelial adenomatoid hamartoma in olfactory clefts. Results All patients were associated with sinusitis, and 23 had sinonasal polyps, 1 had papilloma. On nonenhanced CT, the OC lesions with the OC widening were isodense to gray matter in all cases, and the lesions caused the adjacent bony expansion and absorption rather than erosion; 15 cases were bilateral diseases and 14 were unilateral;The olfactory clefts width of the case patients and the control patients were (1.03±0.24) cm, (0.71± 0.17) cm, respectively. There was statistically significant difference (t=4.963, P<0.01) for the olfactory clefts width between the case patients and the control patients, and there was no significant difference (t=1.640, P>0.05) for the total nasal distance, and was significant difference(t=6.029,P<0.01)in the ratio of OC to the total nasal distance between the two groups. On T1WI, the disease appeared isointense in 6 patients and slightly hypointense in 2 patients compared with gray matter. On T2WI, the lesions revealed heterogeneous isointense in all patients. Regular cribriform pattern was found on MR T2WI and enhanced TlWI. Conclusions The unilateral or bilateral olfactory cleft opacification in chronic sinusitis patients with or without sinonasal polyposis, with involved OC widening and the adjacent bony walls compressed and remodeled may highly suggests the presence of REAH in the OC. The lesions showed inhomogeneous isointense signal on T2WI images, regular cribriform pattern enhancement are typical imaging feature of this entity.

Objective To explore the reconstructive methods of benign bone tumor defects in proximal joint. Methods Operative treatment was performed in 11 cases with benign bone tumor defects in proximal joint, a-mong whom 4 cases were treated by curettage,cauterization of wall and bone grafting,3 cases were treated by filling branch through segment fibula transplantation with vascular,2 cases were treated by fibula head transplantation with vascular to reconstruct the glenohumeral joint and rediocarpal joint, and 2 cases were treated by artificial joint re-placement. Results The follow-up ranged from 1.5 to 6 years. All of cases got excellent bone unioned without re-lapse and 2 cases of them developed with little limitation in joint function. Conclusion The importance and recon-structive achievement of fibula transplantation in treating the benign bone tumor defects in proximal joint should be thought fully. The indication of artificial tumor prosthesis replacement should be strictly mastered.

BACKGROUND: It has been believed mesenchymal stem cells (MSCs) play a role in treatment through paracrine mechanism. Various side effects such as embolism, tumorigenesis and immunological reaction caused by direct injection of MSCs can be avoided by extracting MSC lysate. However, there is a larger difference in current collection methods and standards of MSC lysate. OBJECTIVE: To compare repeated freeze-thaw and ultrasonication for the collection of lysate of MSCs. METHODS: Adipose-derived mesenchymal stem cells (ADMSCs) were isolated from the abdominal subcutaneous fat of healthy individuals, and purified with adherence screening method, followed by in vitro amplification using fetal bovine serum medium. The common surface makers of these cells were tested by flow cytometry (1×109, 2×109, 4×109/L). Repeated freeze-thaw and ultrasonication were employed for cell cytoclasis at three different densities respectively in saline and double distilled water, and a comprehensive comparison was performed on cytoclasis rate and the content of protein in cell lysate between the two methods. RESULTS AND CONCLUSION: (1) ADMSCs obtained from in vitro isolated human adipose tissue grew in a swirl or radial pattern with a homogenous size and neat arrangement. CD44, CD90, CD105 and other commonly used surface markers were highly expressed. (2) The study for optimization of lysate collection revealed that the higher cell density implicated a longer time for cell wall disruption and cytoclasis, as well as significantly increased cytoclasis rate. (3) BCA protein assay showed that the highest content of protein was obtained in saline solvent using ultrasonication method. Comprehensive analysis on the results leads to a conclusion that ultrasonication method with saline as the solvent is the optimized method for extraction of ADMSCs lysate, and the cell concentration of less than 4×109/L is recommended.

BACKGROUND:Apoptosis of islet cel s is closely related to the long-term hyperglycemia-and hyperlipemia-induced injuries. OBJECTIVE:To observe the effect of Shizidaiping formula on the apoptosis and insulin secretion in MIN6 cel s under the high glucose and lipid environment, and to explore the protective effect of Shizidaiping formula and the related apoptosis mechanism. METHODS:MIN6 cel s were divided into normal, model, melbine, low-, medium-and high-dose Shizidaiping formula groups. The cel activity was examined by cel counting kit-8, the insulin secretion was measured by ELISA, the rate of apoptosis was measured by Annexin V-FITC&PI and the expression levels of MEK1/2, ERK1/2 and p-ERK1/2 were examined by western blot assay. RESULTS AND CONCLUSION:Shizidaiping formula significantly improved MIN6 cel activity under high glucose and lipid condition (P<0.05), decreased early cel apoptosis, increased the level of insulin stimulated by low glucose in cel supernatant (P<0.05), and improved the expression levels of MEK1/2, ERK1/2 and p-ERK1/2 (P<0.05). These results suggest that Shizidaiping formula can protect islet cel s from hyperglycemia and hyperlipemia damage by improving the activity of MIN6 cel s, reducing the insulin secretion and inhibiting the apoptosis of pancreaticβcel s in MIN6 cel s.

Objective:To investigate the therapeutic effects of different surgical methods on papillary thyroid carcinoma of the isthmus (PTCI) and their effects on parathyroid function and thyroglobulin.Methods:Eighty patients with PTCI who underwent treatment in Zhejiang Xin'an International Hospital from January 2016 to January 2021 were included in this study. They were randomly allocated to undergo ipsilateral lobectomy with removal of the isthmus combined with ipsilateral central neck lymph node dissection (group A, n = 40) or total thyroidectomy combined with ipsilateral central neck lymph node dissection (group B, n = 40). We compared intraoperative and postoperative conditions, complications, and hypoparathyroidism between the two groups. We also compared serum thyroglobulin level measured before and 3 days after surgery between the two groups. Results:Operative time was significantly shorter in group A than in group B [(78.95 ± 13.52) minutes vs. (104.23 ± 27.38) minutes, t = -5.23, P < 0.05]. Intraoperative blood loss was significantly less in group A than in group B [(52.32 ± 6.59) mL vs. (75.41 ± 9.98) mL, t = -12.21, P < 0.05]. There were no significant differences in voice handicap index and reflux symptom index scores between the two groups (both P > 0.05). The incidence of complications was significantly lower in group A than in group B (10.00% vs. 30.00%, χ2 = 5.00, P < 0.05). The incidence of hypoparathyroidism was significantly lower in group A than in group B (7.50% vs. 27.50%, χ2 = 5.54, P < 0.05). At 3 days after surgery, serum thyroglobulin level was significantly lower in group A than in group B [(0.82 ± 0.17) μg/L vs. (1.26 ± 0.23) μg/L, t = -9.73, P < 0.05]. Conclusion:Ipsilateral lobectomy with removal of the isthmus combined with ipsilateral central neck lymph node dissection is more effective on PTCI than total thyroidectomy combined with ipsilateral central neck lymph node dissection. The former has little effect on parathyroid function and can reduce serum thyroglobulin level. The study is highly innovative and scientific.

Objective To explore the influence of the killer cell immunoglobulin like receptor (KIR) gene polymorphism on cytomegalovirus (CMV) infection and pathogenesis after hematopoietic stem cell transplantation (HSCT).Methods The KIR genotype was determined by sequence-specific primer polymerase chain reaction (PCR-SSP) in 138 pairs of donors and recipients before HSCT during October,2005 and May,2011.Posttransplant monitoring for CMVpp65 antigen was performed by indirect immune histochemically assays since week 2 after transplantation.The differences between CMV positive group and negative group,inhibitive and active KIR of donors and recipients,and KIR haplotype frequency of donors and recipients were analyzed.Results There were no significant differences in frequency of KIR gene and haplotype AA,AB,BB between the donors and recipients.The frequencies of 2DS2 and 2DS4 * 003-007 of donors in CMV positive group were obviously lower than those in CMV negative group with significant differences(8％ vs 16％,P =0.0420;3％ vs 13％,P =0.0050).There was no significant difference in KIR gene between CMV positive group and CMV negative group.The CMV infection rates of haplotype AA,BB,AB donors were 64.38％,36.84％ and 50.00％,while CMV infection rates of haplotype AA,BB,AB recipients were 53.73％,46.15％ and 51.72％,respectively.The CMV infection rate was higher in the patients received KIR haplotype AA donor than in those received KIR haplotype BB donor (36.84％ vs 64.38％,P =0.0299).2DS4 * 003-007 and haplotype BB of donor were found associated with CMV infection in multifactor analysis.Conclusion KIR genotypes of donors are associated with CMV infection after HSCT.