BACKGROUND:Colagen hydrogel provides good matrix support for hepatocyte growth and tissue reconstruction, and the colagen-based engineered tissue is easy to merge the growth and form integrated tissue. OBJECTIVE:To improve the thickness of engineered hepatic tissue by dissociating hepatocytes/colagen hydrogel composite into smal hepatic units that accumulate in the subcutaneous cavity. METHODS:Freshly isolated hepatocytes from rats were mixed with colagen hydrogel to establish hepatocytes/colagen hydrogel composite. The hepatocytes/colagen hydrogel composite was dissociated into smal hepatic units after being cured. The undissociated hepatocytes/colagen hydrogel composite was taken as a control. Six Spraque-Dawley rats were enroled. Three of them were subjected to a two-thirds partial hepatectomy to induce liver regeneration. Dissociated and undissociated hepatocytes/colagen hydrogel composites were implanted into the bilateral inguinal subcutaneous cavity. Dissociated and undissociated hepatocytes/colagen hydrogel composites were implanted into the bilateral inguinal subcutaneous cavity of the other three rats. At the 7th day after transplantation, engineered hepatic tissue formation was evaluated using hematoxylin-eosin staining, immunohistochemical staining and India ink perfusion methods. RESULTS AND CONCLUSION:The grafts in these two groups al formed vascular engineered hepatic tissue in the subcutaneous cavity, but after the smal hepatic units merged, a large piece of vascular engineered hepatic tissue formed. The hepatic tissue thickness was up to 4 mm. The whole piece of implanted liver grafts only formed smal pieces of hepatic tissues, with only several layers of cels. Immunohistochemistry staining confirmed that the hepatocytes in vascular engineered hepatic tissue had the characteristics and functions of natural hepatocytes. Partial hepatectomy experiment showed that engineered hepatic tissue had the ability to respond to regenerative stimulus of partial hepatectomy. These results show that dissociating the hepatocytes/colagen hydrogel grafts into smal units that accumlate in the subcutaneous cavity can increase the thickness of the engineered hepatic tissue.

OBJECTIVE: To investigate the characteristics and influencing factors leading to sudden cardiac death (SCD) of incarcerated prisoners. METHODS: Seventy-five SCD cases of prisoners between 2000 and 2013 in Henan province were collected, and environment, psychological and physical factors were retrospectively analyzed. Combined with histopathological results, specific factors of SCD were also studied. RESULTS: In the 75 cases, 21 cases (28%) had definite chronic past medical histories, and 75 cases (100%) had cardiovascular disease confirmed by autopsy. CONCLUSION Due to presence of the potential cardiac diseases, special incarcerated environment, psychological stress, and body-restraint might be the precipitating factors in SCD of those prisoners.
Autopsy ; Cardiovascular Diseases ; Death, Sudden, Cardiac ; Environment ; Humans ; Mental Disorders ; Prisoners ; Retrospective Studies ; Stress, Psychological

Here we discuss whether N terminal sequencing is appropriate as one of the conventional control methods for monoclonal antibody products. We determined the N terminal sequences of two monoclonal antibody products targeting two antigens separately with both Edman degradation and mass peptide spectrometry. We also identified the characteristic peptide fragments with mass spectrometry. Furthermore, we analyzed their heterogeneity with ion exchange chromatography, capillary zone electrophoresis and Imaged Capillary Isoelectric Focusing. Edman degradation method showed that the N terminal 15 amino acids of heavy and light chains of the two monoclonal antibodies were identical. Peptide mass spectrometry demonstrated that T1 peptide fragments of heavy and light chains of the two antibodies were also the same. But in contrast, peptide mapping and the three analytical methods for heterogeneity analysis could effectively identify and differentiate the two antibodies. The N terminal sequences of two monoclonal antibodies are identical because the number of framework sequences of humanized or human monoclonal antibodies is relatively limited, so whether N terminal sequencing analysis could be regulated as one of the practical control methods should be carefully discussed. Our work also proves that the above analytical methods could combinatorially applied to the identification of monoclonal antibody products, and are more objective compared to N terminal sequencing.
Amino Acid Sequence ; Antibodies, Monoclonal ; isolation & purification ; Chromatography, Ion Exchange ; Humans ; Isoelectric Focusing ; Mass Spectrometry ; Peptide Mapping ; Peptides ; Sequence Analysis, Protein ; methods

Objective To analyze the differences of size and charge heterogeneities between origi-nal humanized anti-TNF-αantibody and four similar biotherapeutic products ( SBP ) .Methods The size exclusion chromatography ( SEC-HPLC ) and weak cation exchange chromatography ( WCX-HPLC ) were used to analyze the size and charge heterogeneities , respectively.Carboxypeptidase B (CpB) treatment was employed to analyze the source of charge heterogeneity of the antibody products .Results Four SBPs showed the same pattern with the originator in SEC-HPLC, and no significant difference with the percentage of mono-mer was observed .The percentages of the aggregates of SBP-3 and SBP-4 were a little higher than those of the originator .The charge distribution of SBPs was significantly different from the originator ′s, especially in the basic region .The results from the samples treated with CpB indicated that the difference of charge distri -bution in the basic region might be caused by the C-terminal lysine variants .Conclusion Four SBPs showed similar size heterogeneity with the originator , but significant differences with charge heterogeneity were observed among them .The study suggested that more attention should be paid to the charge heterogene -ity analysis of the biosimilar products .

Objective To compare the capability of capillary electrophoresis-sodium dodecyl sul-fate ( CE-SDS) and size exclusion-high performance liquid chromatography ( SE-HPLC) for analysis of size heterogeneity of monoclonal antibody products .Methods The size heterogeneity of one humanized anti-VEGF monoclonal antibody was analyzed by using non-reduced and reduced CE-SDS, and conventional , de-natured and denatured reduced SE-HPLC.Results The percentage of aggregates detected by non-reduced CE-SDS (0.82%±0.01%) was equal to that by using denatured SE-HPLC (1.05%±0.02%), but it was significantly lower than that by using conventional SE-HPLC analysis (5.08%±0.10%).With regard to fragments analyzed with non-reduced antibodies, its percentage was (7.12±0.04)% measured by non-re-duced CE-SDS analysis that was significantly higher than that by conventional SE -HPLC analysis (0.02%± 0.01%) and denatured SE-HPLC analysis (0.62%±0.01%).Using reduced antibodies , the percentage of fragments was (3.19±0.50)%tested by reduced CE-SDS analysis that was significantly higher than that by using denatured reduced SE-HPLC analysis (0.07%±0.01%).Conclusion Conventional SE-HPLC was more objective than CE-SDS for content analysis of aggregates , as both the covalent and non-covalent forms of aggregates could be detected .Non-reduced CE-SDS could demonstrate the content of clips , while reduced CE-SDS showed the degraded fragments .Therefore, CE-SDS had an advantage over conventional SE-HPLC for content analysis of fragments .The use of the two analytical methods in combination provided solid techni-cal supports for the quality control of size heterogeneity of monoclonal antibodies .

Objective To explore how to determine the designated hospitals for critical illness scientifically and reasonably.Methods Analyzing the choice of medical providers by inpatients with critical illness,by means of the database of NRCMS in 2009～2010 in one county,Guangxi province.And analyzing the current policies on the basis of general principals used in health policy analysis.Results The choices of medical providers made by inpatients with critical illness are influenced by various factors.Choice of such hospitals should only be dependent on scientific and reasonable determination of patients' reasonable medical needs,instead of on the medical competency of the hospitals only.Conclusion Maximal cost-effectiveness should prevail,be it the class-1 demand of complete healing which is highly dependent on medical technology,or class-2 demand requiring repeated hospitalizations and not highly dependent on medical technology.Reasonable designation of hospitals for critical illness should be based on reasonable medical needs of patients,instead of medical competency of hospitals only.

The biological activity of ADCC by anti-CD20 monoclonal antibody was determined by BioGlo™ Luciferase Assay System using Jurkat/NFAT-luc+FcγRIIIa cell line as effector cell and WIL2-S cell line as target cell. The developed method was verified for specificity, precision and accuracy. Anti-CD20 monoclonal antibody showed a dose-response mode by the developed method, and the determination result complied with the following four-parameter equation: y = (A-D)/[1 + (X/C)(B)] + D. The optimized parameters of the method were determined including the antibodies diluted concentration (18,000 ng·mL(-1)), dilution rate (1:5), the ratio of effector cell and target cell (6:1), and induction time (6 h). The values of eight independent tests have passed a statistical test for curve regression analysis, linear or parallelism, which showed the method possessed good specificity. Four different dilute groups of recovery rates sample were determined for 3 times, and the result showed mean relative potencies of (44.39±3.93)%, (72.74±2.78)%, (128.28±7.01)% and (168.19±2.70)% respectively, with a variation coefficient of less than 10%, and the recoveries of (88.78±7.85)%, (96.99±3.70)%, (102.63±5.61)% and (112.12±1.80)% respectively. A novel reporter gene method for determination of biological activity of ADCC by anti-CD20 monoclonal antibody was successfully developed, which showed strong specificity, good reproducibility and high accuracy, and might be used routinely.

Objective To investigate the clinicopathology and immunophenotype of primary non- Hodgkin lymphoma(NHL)of the female genital system,and to analyze the prognosis of such tumors. Methods Clinicopathologic features of 43 cases of primary NHL of the female genital system were studied retrospectively,with the histological classification based on the Classification of Haematopoietic and Lymphoid Tumors(WHO,2001).Immunochemistry technique,in-situ-hybridization and polymerase chain reaction methods were used to detect the immunophenotype,epstein barrvirus(EB)virus infection status and immunoglobulin heavy chain gene rearrangement,respectively.Results(1)Primary lesions:there were 24 cases of lymphoma originating in the ovary,3 cases in the endometrium,10 cases in the cervix,2 cases in the vagina and 4 cases in the vulva.(2)Staging:12 cases(28%)were in stage Ⅰ,9 cases (21%)in stage Ⅱ,and 22 cases(51%)in stage Ⅲ.(3)Histological classification:37 cases(86%)were diffuse large B cell lymphoma(DLBCL),3 cases were Burkitt lymphoma and the remaining 3 cases were unspecified peripheral T-cell lymphoma according to biopsy,immunophenotype analysis,in-situ- hybridization technique and IgH gene rearrangement detection.(4)Prognosis analysis:increase in the level of lactic acid dehydrogenase,stage Ⅲ,DLBCL and single operation suggest poor prognosis.Conclusions Establishment of the diagnosis of primary NHL of the female genital system is based on biopsy, immunophenotype analysis,in-situ-hybridization technique and IgH gene rearrangement detection,which play important roles in diagnosis and differential diagnosis of the tumor.Combined therapy is the first choice of therapeutic regimens.

Objective To explore the application of B-mode intra- and post-operative ultrasound in patients with traumatic brain injuries. Methods B-mode intra- and post-operative ultrasound were performed 35 times in 24 patients, including 7 times of intra-operation in 7 patients and 28 times of post-operation in 17 patients. Results During operation epidural hematomas in 3 cases as well as intracranial hematomas in 2 cases, subdural hematomas in 1 case, diffused brain swelling in 1 case were found. After operation epidural hematomas were examined in 4 cases as well as subdural hematomas in 1 case, delayed intracranial hematomas in 6 cases, scattered contusion and laceration of the brain in 12 cases, acute obstructive hydrocephalus in 1 case, and subdural hydroma in 4 cases. Six cases underwent operations again because of delayed hematomas, contusion and laceration of the brain. B-mode ultrasound was compared with CT scan and the total coincidence rate was 85.4%.Conclusions B-mode ultrasound is helpful to examine delayed hematomas and other emergent complications. It may save time and may improve rescue rate and has a good coincidence with CT scan. B-mode ultrasound may be widely used as a routine way in neurotrauma ICU.

Objective To study the relationship between activity of thorax and each spinal intervertebral angle in patients with ankylosing spondylitis. Methods Each spinal intervertebral angle of 41 patients with ankylosing spondylitis were measured by Spinalmouse in different postures. And the activity of thorax was measured. Correlation between activity of thorax and shape of spinal were analyzed. Results The activity of thorax positively correlated with the entire lumbar spinal column in flexion (P<0.01), as well as the intervertebral angle of L1/2, L2/3 and L4/5 in flexion (P<0.05), but negatively correlated with the intervertebral angle of L1/2 and L2/3, curvature of the entire lumbar spinal column in upright and the intervertebral angle of L1/2, L3/4, curvature of the entire lumbar spinal column in extension. Conclusion There was a significant relation between activity of thorax and lumbar vertebra motor ability.