BACKGROUND:Low toxicity of Genipin has certain species and cellspecificity. Biocompatibility of Genipin cross-linked type I colagen with human adipose-derived stem cels is essential for construction of
tissue-engineered adipose.
OBJECTIVE:To investigate the bbiocompatibility of Genipin cross-linked type I colagen with human adipose-derived stem cels.
METHODS:Human adipose-derived stem cels were isolated and cultured to the third generation, and the cels were seeded on Genipin cross-linked type I colagen scaffold. MTT assay was used to evaluate the adhesion and proliferation of cels on the scaffold, and the toxic effects of Genipin cross-linked type I colagen on human
adipose-derived stem cels. Optical microscopy and scanning electron microscopy were utilized to observe the adhesion and growth process of human adipose-derived stem cels on the scaffold as wel as the morphological changes of cels.
RESULTS AND CONCLUSION:Human adipose-derived stem cels could adhere to the scaffold immediately
after seeded and increase gradualy on the scaffold, with the average adhesion rate of 86.5%. Optical microscopy and scanning electron microscopy showed that human adipose-derived stem cels adhered wel on the scaffold. The cels increased gradualy over time, and could migrate into the scaffold, and distribute evenly with the passage of time when observed with optical microscopy. The result showed Genipin possesses very low cytotoxicity to the cels, and the outstanding biocompatibility is found between the cels and scaffoldin vitro after cross-linked with Genipin.

BACKGROUND:Based on the original advantages of silk fibroin, positive charged water-soluble chitosan modified silk fibroin is modified on surface and could improve celladhesion on the scaffolds. OBJECTIVE:To verify the biocompatibility of chitosan-modified silk fibroin with human adipose-derived stem cells (hADSCs), and feasibility of constructing tissue engineered adipose in vitro. METHODS:The hADSCs at passage 3 were seeded on chitosan-modified silk fibroin at the concentration of 1×107/L, as the experiment group;at the same cellconcentration, hADSCs were seeded in 96-wel plates as the control group. MTT tests were performed to evaluate the adhesion, growth and proliferation of hADSCs on chitosan-modified silk fibroin. Then hADSCs were implanted on the chitosan-modified silk fibroin scaffolds at the concentration of 1×109/L. The hADSCs seeded onto chitosan-modified silk fibroin complexes were respectively cultured with adipogenic differentiation medium and ordinary high-glucose DMEM. The complexes were stained with oil red O, and detected with RT-PCR after cultured 14 days. RESULTS AND CONCLUSION:The hADSCs adhered to and proliferated on the scaffolds. After cultured with adipogenic differentiation medium for 14 days, oil red O staining demonstrated that there were amount of mature adipocytes on the scaffold. The peroxisome proliferator activated receptorγ2 was positively expressed. The chitosan-modified silk fibroin possessed excellent biocompatibility in vitro. The co-cultured hADSCs could be induced to mature adipocytes successful y.

ObjectiveTo investigate the clinical effect and safety of sofosbuvir (SOF)-ledipasvir (LDV) in the treatment of patients with HCV genotype 6a chronic hepatitis C (CHC). MethodsA total of 63 patients with HCV genotype 6a CHC who visited Department of Infectious Diseases, Henan Provincial People’s Hospital and Nanfang Hospital, from October 2014 to December 2016 were enrolled in this prospective observational study. They were divided into SOF-LDV group (treated with SOF-LDV for 12 weeks) and PR group (treated with pegylated interferon combined with ribavirin for 24 weeks). HCV RNA was measured during treatment and follow-up, and virologic response was evaluated. The chi-square test was used for comparison of categorical data between two groups, and the Mann-Whitney U test was used for comparison of continuous data between two groups. ResultsThere were no significant differences between the PR group and the SOF-LDV group in rapid virologic response rate (85.3% vs 100%, P＞0.05) and virologic response rate at the end of treatment (94.1% vs 100%, P＞005). The SOF-LDV group had a significantly higher sustained virologic response rate than the PR group (96.4% vs 73.5%, χ2=438, P=0.036). The PR group had a significantly higher incidence rate of adverse events than the SOF-LDV group(χ2=754,P=0006). During follow-up, one patient with liver cirrhosis in the SOF-LDV group developed small hepatocellular carcinoma, while no patient in the PR group developed liver cancer at the end of follow-up. ConclusionSOF-LDV for 12 weeks is safe and effective in the treatment of HCV genotype 6a CHC, but liver cancer should be closely monitored in patients with liver cirrhosis.

OBJECTIVETo investigate the effects of adefovir dipivoxil (ADV) on blood phosphorus metabolism in patients with chronic hepatitis B (CHB).

METHODSPatients with hepatitis B surface antigen (HBsAg)-positive CHB were treated with ADV alone, ADV combined with interferon (IFN), or ADV combined with lamivudine (LAM). Changes in levels of calcium, phosphate, urea, and creatinine were assessed at treatment weeks 4, 12, 24, 48, 72 and 96. Statistical analysis was carried out with SPSS 16 software; influential factors were analyzed by ANOVA and non-conditional logistic regression analysis.

RESULTSDuring the course of treatments, 32 (42.6%) of the patients presented with low phosphorus. The highest incidence of low phosphorus was found to have occurred at treatment week 24 (25.0%, 27.5% and 36.4% respectively, with no statistical difference between three groups, x2=0.225, P>0.225). Patients with hypophosphatemia did not show a significant difference in serum phosphorus levels from the other patients (F=1.853, P=0.169). Logistic regression showed a correlation between low phosphorus and sex (x2=7.876, P<0.05), age (t=2.479, P<0.05), and serum creatinine (t =-2.256, P<0.05), but not with blood urea nitrogen or blood calcium (P>0.05).

CONCLUSIONADV antiviral treatment can decrease the blood phosphorous levels of CHB patients, particularly over extended time of treatment, and the occurrence of low phosphorus is more common than of mild phosphorus decrease.Male and elderly patients may be at greater risk of this complication. The incidence and severity of low phosphorus is not significantly different for the different ADV-based treatment regimens.

Adenine ; analogs & derivatives ; Aged ; Antiviral Agents ; Creatinine ; Drug Therapy, Combination ; Hepatitis B, Chronic ; Humans ; Interferons ; Lamivudine ; Male ; Organophosphonates ; Phosphorus

Objective: To investigate the effect of hepatitis B core antigen (HBcAg) in promoting the invasion of hepatitis B virus (HBV)-related hepatocellular carcinoma cell line HepG2.2.15 and the role of Toll-like receptor 4 (TLR4) in the mechanism. Methods: TLR4 mRNA and protein expression in HepG2 cells and HepG2.2.15 cells was measured by reverse transcription real-time PCR and Western blot analysis, respectively. HepG2.2.15 cells were transfected with TLR4 specific small interfering RNA (siRNA) to silence TLR4 expression, and stimulated by recombinant HBcAg in culture. The invasion of cells was measured by Transwell invasion assay. The expression of TLR4 signaling pathway-related proteins in the cultured cells and proinflammatory cytokines in the supernatant was also determined. The student’s t-test, one-way ANOVA, and SNK-q test were used for statistical analysis. Results: TLR4 mRNA and protein expression in HepG2.2.15 cells was significantly higher than that in HepG2 cells. TLR4 siRNA transfection remarkably down-regulated TLR4 expression in HepG2.2.15 cells. Inhibiting TLR4 expression and/or HBcAg stimulation did not affect the proliferation of HepG2.2.15 cells. However, HBcAg stimulation significantly increased the invasion ability of HepG2.2.15 cells and the secretion of proinflammatory cytokines [including interferon (IFN)-γ, interleukin (IL)-6, IL-8, and tumor necrosis factor (TNF)-α]. Inhibiting TLR4 expression significantly reduced HBcAg-induced cellular invasion. Meanwhile, HBcAg stimulation elevated the expression of MyD88 and TRIF in HepG2.2.15 cells. TLR4 silencing inhibited HBcAg-induced increase in the expression of MyD88, while it showed no significant impact on TRIF expression. Conclusion HBcAg can promote the invasion of HepG2.2.15 cells. The TLR4/MyD88 signaling pathway may be involved in this process by inducing the expression of proinflammatory cytokines.

Objective To explore the correlation between adiponectin and liver fibrosis in patients with chronic hepatitis B virus infection.Methods 60 patients with chronic hepatitis B were divided into mild liver fibrosis group (n =28),severe liver fibrosis group (n =17)and cirrho-sis group (n =15),and 20 healthy volunteers were designed as the control group.The serum adiponectin level was detected in all the groups.Results There were significant differences of serum adiponectin levels among the four groups (P <0.05),and there was a positive correlation be-tween serum adiponectin level and liver fibrosis grade (r =0.967,P <0.05).The expression rate of liver tissue adiponectin in cirrhosis group was 93.33%,which was significantly higher than the mild group (14.29%)and severe fibrosis group (41.18%)(P <0.05).Conclusion Adiponectin is closely related to the liver fibrosis in patients with chronic hepatitis B virus infection.

Objective To explore the correlation between adiponectin and liver fibrosis in patients with chronic hepatitis B virus infection.Methods 60 patients with chronic hepatitis B were divided into mild liver fibrosis group (n =28),severe liver fibrosis group (n =17)and cirrho-sis group (n =15),and 20 healthy volunteers were designed as the control group.The serum adiponectin level was detected in all the groups.Results There were significant differences of serum adiponectin levels among the four groups (P <0.05),and there was a positive correlation be-tween serum adiponectin level and liver fibrosis grade (r =0.967,P <0.05).The expression rate of liver tissue adiponectin in cirrhosis group was 93.33%,which was significantly higher than the mild group (14.29%)and severe fibrosis group (41.18%)(P <0.05).Conclusion Adiponectin is closely related to the liver fibrosis in patients with chronic hepatitis B virus infection.

Objective:To investigate the clinical characteristics of patients with aspergillus spondylitis, and to provide reference for timely diagnosis and treatment.Methods:The clinical manifestations, imaging performance, laboratory examination results, diagnosis and treatment outcomes of six patients with confirmed aspergillus spondylitis in Department of Infectious Diseases, Henan Provincial People′s Hospital during April 30, 2015 and May 1, 2020 were retrospectively analyzed.Results:The main manifestations of six patients were fever and neck pain or low back pain. The time from the onset of clinical manifestations to diagnosis was more than two months to 14 months. Spine magnetic resonance imaging (MRI) showed long T1 and T2 signals on vertebral body, high pressure lipid signal, obvious enhanced scan enhancement, and paravertebral abscess formation might be presented. Among the six patients, C-reactive protein increased in four patients, erythrocyte sedimentation rate increased in five patients, β-D-glucan test (G test) increased in three patients, galactomannan antigen test (GM test) increased in four patients. Six patients with aspergillus spondylitis were all confirmed by biopsy of diseased tissue for fungal smear, tissue culture or metagenomics next generation sequencing. After treatment with voriconazole or itraconazole, five patients recovered and one patient was still under treatment.Conclusions:The clinical manifestations and imaging examination of patients with aspergillus spondylitis are nonspecific. Peripheral blood G test and GM test need to be combined for diagnosis. The diagnosis depends on tissue puncture pathology examination, and the metagenomics next generation sequencing is needed if necessary.

Objective:To study the use of preS1-tp fusion protein as a novel vector to mediate the entry of small interfering RNA (siRNA) targeting the carboxy-terminal nuclear localization signal (NLS) region of hepatitis B virus (HBV) core protein into HBV-infected hepatocytes, and to further explore the HBV replication inhibition and covalently closed circular DNA synthesis.Methods:HepG2.2.15 cells expressing the human sodium taurocholate co-transporting polypeptide were established on the basis of lentivirus infection system. siRNA against HBV NLS region was designed and synthesized. PreS1-tp fusion protein expression and purification was observed to test its ability to cell entry and DNA binding. NLS siRNA were delivered into HepG2.2.15- sodium taurocholate co-transporting polypeptide cells by preS1-tp fusion protein as a vector to observe the effects of NLS siRNA on HBV replication and covalently closed circular DNA levels. Analysis of variance was used for comparison between multiple groups, and the measurement data differences between groups were analyzed by t-test.Results:HepG2.2.15-sodium taurocholate co-transporting polypeptide cell line was successfully constructed. Screened synthetic HBV NLS siRNA had significantly inhibited HBV replication. The preS1-tp fusion protein was expressed and purified on a large-scale. The fusion protein as a vector for HBV NLS siRNA had targeted delivery. The result showed that the fusion protein had effectively targeted siRNA to Hepg2.2.15-sodium taurocholate co-transporting polypeptide cell, which not only had effectively inhibited the expression of HBV mRNA, HBsAg and HBeAg, but also had significantly reduced the levels of HBV DNA and covalently closed circular DNA.Conclusion:The preS1-tp fusion protein constructed in this study uses the dual functional characteristics of preS1 binding to hepatocyte HBV receptor, and tp binding to nucleic acids, and targets HBV NLS siRNA against HBV-infected cells and block rcDNA from being transported to nucleus. siRNA plays a role in inhibiting HBV replication and covalently close circular DNA synthesis, providing a new strategy for the treatment of chronic hepatitis B caused by HBV infection, and a new research perspective for the complete elimination of HBV from the body.

OBJECTIVE To evaluate the intervention effect of Caulis sinomenii compatible with prepared Aconiti Lateralis on bone destruction in rheumatoid arthritis （RA）model rats ，and to investigate its mechanism. METHODS Totally 40 SD rats were randomly divided into blank group ，model group ，positive control group （indomethacin 0.013 5 g/kg）and C. sinomenii compatible with prepared Aconiti Lateralis group （C. sinomenii 1.08 g/kg+prepared Aconiti Lateralis 1.35 g/kg）according to body mass ，with 10 rats in each group. Except for the blank group ，all the other groups made RA rat models by injecting type Ⅱ bovine collagen. Rats in each group were given corresponding drugs or distilled water intragastrically. The general information ，body weight ，foot swelling and arthritis index （AI）scores of rats in each group were recorded. After the 30th day of administration ，the changes of ankle bone in rats were detected by small animal CT machine. The levels of inflammatory factors [interleukin- 31（IL-31），IL-25 and IL- 3] and chemokines [receptor activator of nuclear factor κB ligand（RANKL），receptor activator of nuclear factor κB （RANK）and osteoprotegerin （OPG）] in serum were detected by enzyme-linked immunosorbent assay. Pathological indexes of rat ankle joint were observed by HE staining. Immunohistochemical method was used to detect the expression of RANKL ，RANK and OPG in synovial tissue of rat ankle joint. RESULTS Compared with blank group ，the mental state of the model group was weak ， the activity decreased significantly ，the hair lost luster ，and the body weight decreased significantly on the 12th to 30th days （P＜ 0.05 or P＜0.01）；the swelling degree of the foot was significantly increased and the AI score was significantly increased on the 12th to 30th days（P＜0.01）；the ankle joint in model group had rough surface ，obvious tissue damage and serious bone erosion ； serum levels of IL- 31，IL-25，IL-3，RANKL and RANK were increased significantly ，while the level of OPG was decreased significantly （P＜0.01）； the expression of RANKL and RANK in synovium of ankle joint increased significantly ， while the expression of OPG decreased significantly （P＜0.01）. Compared with model group ，the above indexes of administration groups were improved to varying degrees ，and most of the differences were statistically significant （P＜0.05 or P＜0.01）. CONCLUSIONS By inhibiting the RANKL/RANK/OPG signaling pathway ，C. sinomenii compatible with prepared Aconiti Lateralis can inhibit the excessive proliferation of osteoclasts and restore the balance of bone metabolism so as to play a role in protecting bone joints and treating RA.