Objective:To fabricate the composite scaffolds for bone regeneration with silk fibroin (SF), bacterial cellulose nanofibers (BCNR) and hydroxyapatite (HAp) and evaluate their osteogenic activity.Methods:HAp particles, BCNR and bone morphogenetic protein-2 (BMP2) were added into SF aqueous solution in turn, poured into molds of different sizes after being mixed evenly and processed at -25 ℃ for 24 hours to obtain frozen molds, and the composite scaffolds were frozen-dried by freezing-drying machine. The composite scaffolds with different mass ratios of SF and BCNR were divided into groups A (2∶1), B (4∶1) and C (6∶1), and the inactive composite scaffolds without BMP2 fell into group D. The surface morphology and pore structure of the scaffolds were detected by scanning electron microscopy. The porosity of the scaffolds was measured by mercury intrusion porosimeter. The stress-strain curve was obtained by using the universal material testing machine to compress the scaffolds, with which their compressive strength and Young′s modulus were analyzed. Immortalized mouse embryonic fibroblasts (iMEF) were inoculated on the composite scaffolds of group A, B, C and D. At 4 and 8 days after cell inoculation, the proportion of alive and dead cells in each group was detected by cell survival/death staining; the cell counting kit-8 (CCK-8) was used to detect cell proliferation activity in each group; the positive staining cells were detected in each group by alkaline phosphatase (ALP) staining; the ALP activity was observed in each group with ALP activity detection. A total of 15 female SD rats were selected to establish osteogenesis models with ectopic muscle bag. The composite scaffolds implanted with different SF/BCNR mass ratios and the inactive composite scaffolds without BMP2 fell into group A′ (2∶1), B′ (4∶1), C′ (6∶1) and D′ respectively, and a sham operation group was set at the same time, with 3 rats in each groups. In the sham operation group, the muscle bag and skin were sutured without scaffold implantation after the incision of skin, the blunt separation of the quadriceps muscle, and the formation of muscle bag in the muscle. In the other four groups, the corresponding scaffolds were implanted in the muscle bag and the muscle bag and skin were sutured. X-ray examination was performed at 2 and 4 weeks after operation to observe the osteogenesis in each group. At 4 weeks after operation, the implanted scaffolds and tissue complexes were collected by pathological tissue sectioning, HE staining and Masson staining, and for observing the osteogenesis by in each group. Immunohistochemical staining was also performed on the tissue sections to observe the expression of osteogenic markers type I collagen (COL1) and osteopontin (OPN) in each group.Results:Scanning electron microscopy showed that the lamellar and micropore structures of group B were more regular and uniform than those of groups A and C. The porosity rate analysis showed that the porosity rates of groups B and C were (89.752±1.866)% and (84.257±1.013)% respectively, higher than that of group A [(81.171±1.268)%] ( P<0.05 or 0.01), with the porosity rate of group C lower than that of group B ( P<0.01). The mechanical property test showed that the compressive strengths of groups B and C were (0.373±0.009)MPa and (0.403±0.017)MPa respectively, higher than that of group A [(0.044±0.003)MPa] ( P<0.01), and the Young′s moduli of groups B and C were (7.413±0.094)MPa and (9.515±0.615)MPa respectively, higher than that of group A [(1.881±0.036)MPa] ( P<0.01), with the compressive strength and Young′s modulus of group C higher than those of group B ( P<0.05 or 0.01). The cell survival/death staining showed that the number of dead cells of group B was significantly smaller than that of groups A, C and D at 4 days after cell inoculation, and that group B had the most living cells and the fewest dead cells at 8 days after cell inoculation. The results of CCK-8 experiment showed that at 4 days after cell inoculation, the cell proliferation activity of groups A and B was 0.474±0.009 and 0.545±0.018 respectively, higher than 0.394±0.016 of group D ( P<0.01); the cell proliferation activity of group C was 0.419±0.005, with no significant difference from that of group D ( P>0.05), while the cell proliferation activity of groups A and C were both lower than that of group B ( P<0.01). At 8 days after cell inoculation, the cell proliferation activity of group B was 1.290±0.021, higher than 1.047±0.011 of group D ( P<0.01); the cell proliferation activity of group C was 0.794±0.032, lower than that of group D ( P<0.01); the cell proliferation activity of group A was 1.086±0.020, with no significant difference from that of group D ( P>0.05); the cell proliferation activity of groups A and C was lower than that of group B ( P<0.01). At 4 and 8 days after cell inoculation, ALP staining showed that more positive cells were found in groups A, B and C when compared with group D, and more positive cells were found in group B than in groups A and C. At 4 days after cell inoculation, the ALP activity detection showed that the ALP activity of groups A, B and C was 1.399±0.071, 1.934±0.011 and 1.565±0.034 respectively, higher than 0.082±0.003 of group D ( P<0.01), while the ALP activity of groups A and C was lower than that of group B ( P<0.01). At 8 days after cell inoculation, the cell activity of groups A, B and C was 2.602±0.055, 3.216±0.092 and 2.145±0.170 respectively, higher than 0.101±0.001 of group D ( P<0.01), while the ALP activity of groups A and C was lower than that of group B ( P<0.01). X-ray examination results showed that at 2 weeks after operation, no obvious osteogenesis was observed in the sham operation group, group D′, A′ and C′, while it was observed in group B′. At 4 weeks after operation, obvious osteogenesis was observed in group A′, B′ and C′, with significantly more osteogenesis in group B′ than in the other two groups, while there was no obvious osteogenesis in the sham operation group and group D′. At 4 weeks after operation, the HE staining and Masson staining showed that a large number of uniformly distributed new bone tissue was formed in group B′, while only a small amount of new bone tissue was found locally in groups A′ and C′, and only part of new tissue was found to grow in group D′ with no obvious new bone tissue observed. The maturity of new bone tissue formed in group B′ was higher than that in group A′ and C′. Immunohistochemical staining showed more COL1 and OPN positive staining in group B′ when compared with groups A′ and C′. The expression intensity analysis of COL1 and OPN showed that in groups A′, B′ and C′, the expression intensity of COL1 was 2.822±0.384, 22.810±2.435 and 12.480±0.912 respectively and the expression intensity of OPN was 1.545±0.081, 5.374±0.121 and 2.246±0.116 respectively, with higher expression intensity of COL1 and OPN in groups B′ and C′ than that in group A′ ( P<0.01) and lower expression intensity of COL1 and OPN in group C′ than that in B′ group ( P<0.01). Conclusions:The composite scaffold for bone regeneration is successfully fabricated with SF, BCNR and HAp. The composite scaffold with a mass ratio of SF to BCNR of 4∶1 has uniform pore structure, high porosity, good mechanical properties and biocompatibility, excellent pro-osteogenic properties in vitro, as well as excellent osteo-inductivity and osteo-conductivity.

This paper summarizes the exploration process and academic significance of the academic thought of Zhou Zhongying,a master of traditional Chinese medicine,who took the creation of a new system of TCM pathogenesis theory as the core,and interprets its theoretical connotation.As a pioneer in the construction of higher education textbooks for traditional Chinese medicine,Professor Zhou Zhongying created the outline of TCM internal medicine viscera differentiation,persisted in carrying out innovative research on patho-genesis theory,achieved fruitful academic results,and enriched and developed the academic system of TCM theory.In the clinical di-agnosis and treatment of exogenous febrile diseases and acute and difficult internal injuries,he systematically created new pathogenesis theories such as stasis-heat theory and cancer toxicity theory.Based on this,the legislation of medication can improve the clinical effi-cacy,and it is realized that identifying the pathogenesis is the key link in syndrome differentiation and treatment.In his later years,Professor Zhou Zhongying,guided by the holistic view,proposed the"thirteen pathogenesis"and constructed a new system of TCM pathogenesis differentiation,highlighting the guiding value of complex pathogenesis and the causal chain of pathogenesis elements to complex clinical diseases and syndromes,forming a theory with the idea of"examining syndromes and seeking pathogenesis,activating syndrome differentiation"as its soul.This theory breaks through the rigid thinking of syndrome differentiation and treatment based on a single pathogenesis or fixed syndrome type,reconstructs the theoretical framework of TCM with the idea of holistic view,and is a major academic innovation in modern TCM.

‍Traditionally， the progression from compensated liver cirrhosis to decompensated liver cirrhosis has been considered an irreversible point in the natural history of the disease； however， with the suppression of underlying etiology， cure， and disease regression， this view is challenged by an increasing number of new evidence， and the idea of “recompensation of liver cirrhosis” is gradually being accepted. In recent years， scholars in China and globally have been exploring the specific definition of recompensation of liver cirrhosis and the clinical features of patients. By summarizing the recent studies on recompensation of liver cirrhosis in China and globally， integrating existing views， and analyzing related research evidence， this article points out the main challenges in the field of recompensation at this stage， including the lack of in－depth clinical and basic research， the need to define recompensation in the context of NAFLD， and related ethical issues， in order to provide new directions for future research in this field.

OX40 is a costimulatory receptor that is expressed primarily on activated CD4⁺, CD8⁺, and regulatory T cells. The ligation of OX40 to its sole ligand OX40L potentiates T cell expansion, differentiation, and activation and also promotes dendritic cells to mature to enhance their cytokine production. Therefore, the use of agonistic anti-OX40 antibodies for cancer immunotherapy has gained great interest. However, most of the agonistic anti-OX40 antibodies in the clinic are OX40L-competitive and show limited efficacy. Here, we discovered that BGB-A445, a non-ligand-competitive agonistic anti-OX40 antibody currently under clinical investigation, induced optimal T cell activation without impairing dendritic cell function. In addition, BGB-A445 dose-dependently and significantly depleted regulatory T cells in vitro and in vivo via antibody-dependent cellular cytotoxicity. In the MC38 syngeneic model established in humanized OX40 knock-in mice, BGB-A445 demonstrated robust and dose-dependent antitumor efficacy, whereas the ligand-competitive anti-OX40 antibody showed antitumor efficacy characterized by a hook effect. Furthermore, BGB-A445 demonstrated a strong combination antitumor effect with an anti-PD-1 antibody. Taken together, our findings show that BGB-A445, which does not block OX40-OX40L interaction in contrast to clinical-stage anti-OX40 antibodies, shows superior immune-stimulating effects and antitumor efficacy and thus warrants further clinical investigation.
Mice ; Animals ; Receptors, Tumor Necrosis Factor/physiology* ; Receptors, OX40 ; Membrane Glycoproteins ; Ligands ; Antibodies, Monoclonal/pharmacology* ; Antineoplastic Agents/pharmacology*

Objective The concentration of NE and TNF-α was detected for elucidating the change of them in sera and bronchoalveolar lavage fluid (BALF) of rabbits with acute lung injury interposed by XueBiJing therapy in underwater explosion.Methods Underwater explosion decive was applied to cause acute lung injury of rabbits.The 20 rabbits were randomly divided into two groups which were injury group and XueBiJing therapy group,respectively.The concentration of NE and TNF-α in sera and BALF were detected by ELISA method in 24 hours after bursting.Results The concentration of TNF-α in sera (353.30±166.86 ng/L) of rabbits in therapy group were significantly lower than those in injury group (552.30± 169.64 ng/L;t=2.645,P =0.016).The concentration of NE in sera (63.40 ± 36.09 ng/ml) were lower than that of rabbits in injury group (97.60 ± 36.20 ng/ml;t=2.116,P=0.049).At the same time the concentration of NE in BALF (102.10± 9.50 ng/ml) of rabbits in therapy group were significantly lower than those in injury group (136.70± 13.60 ng/ml;t=6.593,P=0.000).Conclusion The expression of TNF-α in sera and NE in sera and BALF in rabbits with acute lung injury interposed by XueBiJing in underwater explosion were lower than those of injury group.

ObjectiveTo explore regulation effect of miR-340-5p on regulating bone morphogenetic protein 4(BMP4) expression and the differentiation of rat's NSCs.MethodsNSCs of rats were divided randomly into three groups: normal group (Mock),group with nonsense oligonucleotide (Anti-Con) and group with antisense oligonucleotide of miR-340-5p (Anti-miR-340-5p).The qRT-PCR was used to detect the relative expression of miR-340-5p.The expression of NF200 and MAP-2 was detected by immunocytochemical staining and immunofluorescence,respectively.Western blot was used to detect the expression of BMP4 protein.ResultsThe relative levels of miR-340-5p expression were significantly decreased in Anti-miR-340-5p group (0.14±0.01) compared with that of Mock group(1.01±0.17) and Anti-Con group(1.07±0.13) (P<0.01).Immunocytochemical staining indicated that NF200 was positive in cells of Anti-miR-340-5p group.The proportion of MAP2 positive cells was increased in Anti-miR-340-5p group compared with other groups (P<0.05).Western blot showed the increased expression of BMP4 protein in Anti-miR-340-5p group (0.84±0.09) compared with Mock group(0.53±0.04) and Anti-Con group (0.63±0.09) (P<0.05).ConclusionThe miR-340-5p may exert a potential function in regulating differentiation of NSCs into neurons through a negative regulation of BMP4.

Objective To investigate the grading diagnostic value of magnetic resonance spectroscopy and magnetic resonance imaging and gross motor rating scale for children with cerebral palsy.Methods Forty cases of healthy children under 1.0 ～ 2.0 years old and 42 cases of cerebral palsy children under 1.1 ～ 2.0 years old were enrolled in the study,and we compared the inspection reports on MRI and MRS statistical analysis.According to the results of MRI for dividing cerebral palsy,and MRS indexing of NAA/Cr,CHO/Cr,LAC/Cr levels were compared;different types of cerebral palsy by GMFM scores were examined.Results Nuclear magnetic resonance(MRI) and MRS could effectively distinguish typical pathological changes in the brain of children with cerebral palsy,the cerebral NAA,Cr,Cho could appear significant formant when the children detected by MRS,could be quantitatively detected with MRS.The levels of NAA/Cr、CHO/Cr、LAC/Cr of different MRS and MRI degree was different,and the difference of moderate CP and severe CP was significant (P ＜ 0.05).Compared with MRS,the accordant rates of the MRI was not ideal,But gross motor rating scale score suggested that there were differences between the cerebral palsy of the MRI and MRS degree (P ＜ 0.05).Conclusion The magnetic resonance imaging and magnetic resonance spectroscopy and gross motor rating scale can effectively improve the dividing diagnosis of cerebral palsy,and this method for different types of cerebral palsy in children has some guiding value.

Objective To investigate the feasibility, safety and efficiency of endovascular treatment for patients with aorto-bilateral-iliac artery total occlusive disease.Methods A total of 35 patients with aorto-bi-iliac artery total occlusive disease treated with endovascular therapy in Peking Union Medical College Hospital and the First Hospital of Shijiazhuang between Jan 2012 and Dec 2013 were retrospectively analyzed.Results There were 33 males and 2 females, mean age (67 ± 6) years treated during the study period.Technical success rate was 100％.129 bare stents and 4 covered stents were implanted.There were no peri-operative death.Postoperative leg ankle brachial index (ABI) improved significantly (0.86 vs.0.28, P ＜ 0.28).Postoperative complications occurred in 2 patients (5.7％), including brachial artery thrombosis and rupture of external iliac artery post-dilation.The mean follow-up period was 16.5 months (2-28 months).Two patients (5.7％) were lost to follow up.Re-intervention was performed in 3 patients (8.6％) due to reocclusion of the stents.Primary patency was 91％ (30/33) Conclusions Endovascular treatment is effective for aorto-bi-iliac artery total occlusive disease with low complications and acceptable mid-term patent rate.

ABSTRACTOBJECTIVE To establish measuring scale for the pathogen unit of heat?stasis on hemorrhagic stroke and explore the objectivestandard and quantitative method for measuring heat?stasis pathogenesis.METHODS Measuring questionnaire was designed according to the production process of standard scaleand heat?stasis information representation was selected through three rounds of expert consultationso as to establish the entry pool of scale.Related metrical data of hemorrhagic stroke due to heat?stasis was collected on spots with the clinic epidemiology/DME principles for 4 timesand the credibilityvalidity and applicability of scale were assessed.RESULTS By applying the method of subjective evaluation and step screening based on importance evaluationthe original entry pool of scale concerning hemorrhagic stroke due to heat?stasis was estab-lishedwhich included 32 information representationsbelonging to the dimension of blood heatblood stasis and consciousness respectively.And the scale was further optimized after testing the clinical sampleswith 25 items of higher credibility and va-lidity were chosen.CONCLUSION We have preliminarily established a set of measuring tool for the pathogen unit of heat?sta-sis on hemorrhagic strokeand the scale shows that consciousness change is an important representation in distinguishing hem-orrhagic stroke due to heat?stasis from other diseases.

Objective To analyze the influence of vasoactive drugs on hemodynamics in septic shock patients and to explore the guiding value of systemic vascular resistance index (SVRI) for the application of vasoactive drugs .Methods 21 cases of septic shock in ICU from March 2010 to April 2012 were retrospectively analyzed .The PiCCO monitor technique was applied to monitor the he-modynamic parameters including the cardiac index (CI) ,intrathoracic blood volume index (ITBI) ,systemic cenous resistance index (SVRI) and mean arterial pressure(MAP) ,once per 6-8 h .ITBI>850 mL/m2 was taken as the normal criterion .The data of 202 cases meeting the criterion were collected .These cases were divided into two groups according to whether MAP reach 65 mm Hg , the differences of hemodynamic parameters between the two groups were compared .The MAP≥65 mm Hg group was redivided in-to the group 1 ,2 and 3 according to SVRI .Blood characteristics of pressure ,CI and ITBI were compared among these 3 groups .Re-sults The ITBI value in the MAP<65 mm Hg group and the MAP≥65 mm Hg group was higher than the normal value .The CI and SVRI values in the MAP≥65 mm Hg group were higher than those in the MAP< 65 mm Hg group(P<0 .05) ,while the ITBI value had no difference between the two groups .The ITBI values in the three groups with different SVRI all were higher than the upper normal limit ,furthermore ,in the comparisons among 3 groups ,only comparison between the group 1 and the group 3 had sta-tistical difference(P= 0 .47) .When SVRI was lower ,the CI value was higher than the normal value .Along with the increase of SVRI ,MAP increased gradually ,and CI gradually declined .When SVRI was too high ,MAP had no significant increase ,but the CI value was lower than the lower limit .Conclusion The SVRI monitoring by PiCCO can help to guide the application of vasoactive drugs in septic shock patients ,lower SVRI means the declined vascular reaction and higher SVRI indicates the low cardiac function , moreover blood pressure can not be further enhanced by vascular excessive contraction .