Role of HBV DNA polymerase in mediating the immune escape of tumor cells in HBV-related hepatocellular carcinoma / 临床肝胆病杂志

ObjectiveTo determine whether HBV DNA polymerase is associated with T-cell failure and thus mediates the immune escape of HBV-related hepatocellular carcinoma （HCC） tumor cells， and to investigate the specific molecular mechanisms. MethodsLiver cancer cell lines Huh7 and HepG2 stably transfected with HBV DNA polymerase expression plasmid with Flag （Flag-HBV-P） and intercellular adhesion molecule-1 （ICAM1） were co-cultured with Jurkat cells， and MTT assay， qRT-PCR， and ELISA were used to measure Jurkat cell proliferation， activation （CD69 expression）， and secretion of the cytokine IFN-γ. RNA-seq was used to screen for differentially expressed immune-associated molecules between stably transfected cell lines and control cells， and mRNA half-life and protein half-life assays were used to determine the specific levels of the immune-associated molecules that were affected by HBV DNA polymerase. Related websites were used to predict the transcription factors that may bind to the promoter region of this immune-associated molecule， Western blot was used to verify the effect of transcription factors on the immune-associated molecule， and rescue experiment was used to determine whether HBV DNA polymerase affects the expression level of the immune-associated molecule through this transcription factor. The independent-samples t test was used for comparison between two groups. ResultsThe experimental group had significant reductions in Jurkat cell proliferation， activation， and cytokine secretion compared with the control group （all P<0.01）. Compared with the control group， the experimental group （Huh7 and HepG2 cell lines） had significant reductions in the mRNA and protein expression levels of ICAM1 （all P<0.01）. Website prediction identified the ICAM1 promoter and preliminarily highlighted NFKB1， RELA， and STAT3. Compared with the control group， the experimental group （Huh7 and HepG2 cell lines） had a significant reduction in the protein expression level of p65 （all P<0.01）. After p65 overexpression， there was a significant increase in the protein expression level of ICAM1， and after the expression of p65 was reduced， there was a significant reduction in the protein expression level of ICAM1 （all P<0.01）. In the rescue experiment， there was no significant difference in the protein expression level of ICAM1 between the control group and the experimental group after p65 overexpression （all P>0.05）. After the overexpression of ICAM1， there were no significant differences in the proliferation， activation， and cytokine secretion of Jurkat cells between the control group and the experimental group （Huh7 and HepG2 cell lines） （all P>0.05）. ConclusionHBV DNA polymerase downregulates the level of ICAM1 to mediate HCC immune escape by inhibiting the expression of p65 in NF-κB.

Effect of Yinchenhao decoction on renal oxidative stress injury in rats with obstructive jaundice and its mechanism of action based on the nuclear factor erythroid 2-related factor 2 signaling pathway / 临床肝胆病杂志

Objective To investigate the effect of Yinchenhao decoction on renal oxidative stress injury in rats with obstructive jaundice and its association with the regulation of the expression of nuclear factor erythroid 2-related factor 2 (Nrf2) and nuclear translocation. Methods A total of 32 male Sprague-Dawley rats were randomly divided into sham-operation group (S group), model group (O group), low-dose Yinchenhao decoction group (LY group), and high-dose Yinchenhao decoction group (HY group), with 8 rats in each group. For the rats in the S group, the upper common bile duct was isolated without ligation, and for those in the other groups, double ligation of the middle and upper 1/3 of the common bile duct was performed to establish a model of obstructive jaundice. After 7 days, the rats in the LY group and the HY group were given Yinchenhao decoction by gavage at a dose of 6.3 and 18.9 mL/kg, respectively, while those in the S and O groups were given an equal volume of distilled water by gavage every day for 7 consecutive days, and the rats were treated on day 14. ELISA was used to measure the serum levels of total bilirubin (TBil), direct bilirubin (DBil), alanine aminotransferase (ALT), gamma-glutamyl transpeptidase (GGT), blood urea nitrogen (BUN), and creatinine (Cr); spectrophotometry was used to measure the activity of the oxidative stress factors superoxide dismutase (SOD) and malondialdehyde (MDA) in renal tissue; quantitative real- time PCR and Western blotting were used to measure the mRNA and protein expression levels of Nrf2, Kelch-like ECH-associated protein 1 (Keap1), and NAD(P)H quinone dehydrogenase 1 (NQO1) in renal tissue; immunohistochemistry was used to measure observe the nuclear translocation of Nrf2 protein in renal tissue. A one-way analysis of variance was used for comparison of continuous data between multiple groups, and the least significant difference t -test was used for further pairwise comparison within groups. Results Compared with the S group, the O group had significant increases in the levels of TBil, DBil, ALT、GGT, BUN, and Cr, a significant reduction in the activity of SOD, and a significant increase in the level of MDA (all P 0.05). Compared with the S group, the O group had a significant reduction in the positive rate of Nrf2 in cell nucleus in renal tissue ( P < 0.05), and compared with the O group, the LY group and the HY group had a significant increase in the positive rate of Nrf2 in cell nucleus ( P < 0.05). Conclusion Yinchenhao decoction can effectively alleviate renal injury caused by obstructive jaundice, possibly by upregulating the protein expression of Nrf2 in renal tissue and regulating the nuclear translocation of Nrf2 protein, so as to mediate the protein expression of downstream NQO1, regulate oxidative stress response caused by obstructive jaundice, and thereby alleviate renal injury in rats.

Effects of electroacupuncture on postoperative acute lung injury in patients with acute abdomen complicated with abdominal infection / 中华麻醉学杂志

Objective To evaluate the effects of electroacupuncture (EA) on postoperative acute lung injury (ALI) in patients with acute abdomen complicated with abdominal infection.Methods Ninety patients of both sexes with acute abdomen complicated with abdominal infection,with body mass index of 18-30 kg/m2,aged 35-64 yr,of American Society of Anesthesiologists physical status Ⅰ or Ⅱ,scheduled for elective abdominal surgery with general anesthesia,were divided into 3 groups (n =30 each) using a random number table method:control group (group C),EA at acupoint group (group E) and EA at non-acupoint group (group N).Anesthesia was induced with midazolam,propofol,cis-atracurium and sufentanil.The patients were mechanically ventilated after tracheal intubation.Anesthesia was maintained with inhalation of sevoflurane,Ⅳ infusion of propofol and cis-atracurium and intermittent Ⅳ boluses of sufentanil.At 15 min of anesthesia induction,2 h after skin incision and 2,12 and 24 h after tracheal intubation,Hegu,Zusanli and Neiguan acupoints were stimulated for 15 min every time in group E,and the points 1 cm lateral to the acupoints of Hegu,Zusanli and Neiguan were stimulated for 15 min every time in group N,with disperse-dense waves,frequency 2/15 Hz,wave length 0.2-0.6 ms,at a voltage of 1-2 mA.Arterial blood samples were collected at 15 min before induction (T1) and 24 and 48 h after operation (T2 and T3) for blood gas analysis,oxygenation index was calculated,and the development of ALI (oxygenation index ＜ 300 mmHg) was recorded at T2 and T3.The concentrations of club cell protein 16 (CC16),surfactant protein D (SP-D),soluble receptor for advanced glycation end products (sRAGE),interleukin-1 (IL-1),IL-10,and tumor necrosis factor-alpha (TNF-α) in serum were detected by enzyme-linked immunosorbent assay.Results Compared with the baseline at T1,the serum concentrations of CC16,SP-D,sRAGE,IL-1 and TNF-α were significantly increased,and the serum IL-10 concentrations were decreased at T2 in C,E and N groups (P＜0.05).Compared with group C,the serum concentrations of CC16,SP-D,sRAGE,IL-1 and TNF-α were significantly decreased,and the serum IL-10 concentrations were increased at T2 and T3,and AI was increased at T3 in group E (P＜0.05).There was no significant difference in the incidence of ALI at T2 and T3 among three groups (P＞0.05).Conclusion Although EA does not decrease the occurrence of ALI,it mitigates the degree of ALI to some extent in the patients with acute abdomen complicated with abdominal infection.

Potent and conditional redirected T cell killing of tumor cells using Half DVD-Ig

Novel biologics that redirect cytotoxic T lymphocytes (CTLs) to kill tumor cells bearing a tumor associated antigen hold great promise in the clinic. However, the ability to safely and potently target CD3 on CTL toward tumor associated antigens (TAA) expressed on tumor cells remains a challenge of both technology and biology. Herein we describe the use of a Half DVD-Ig format that can redirect CTL to kill tumor cells. Notably, Half DVD-Ig molecules that are monovalent for each specificity demonstrated reduced non-specific CTL activation and conditional CTL activation upon binding to TAA compared to intact tetravalent DVD-Ig molecules that are bivalent for each specificity, while maintaining good drug like properties and appropriate PK properties.

Three-dimensional finite element analysis of lumbosacral reconstruction with a novel four-rod fixation system / 中华骨科杂志

Objective To analysis the biomechanical properties of four-rod fixation system for lumbosacral reconstruction after sacrectomy.Methods Two finite element models of lumbosacral area were established according to pelvic CT images:a subtotal saerectomy model and a total sacrectomy model.The range of motion of L3 vertebral body and max yon Mise stress of instruments of a novel four-rod fixation system were evaluated and compared with those of modified Galveston technique(MGT)under conditions of veaebral body and max von Mise stress of instruments of the four-rod fixation system were evaluated and compared with those of MGT under vertical compression.Results In subtotal sacreetomy model,the range of motion of L3 vertebral body in the four-rod fixation system were 4.2 mm,0.49°,0.49°,0.24°and 0.26°compared to 5.5 mm.0.74°,0.74°,0.27°and 0.30°in the MGT.The max von Mise stress of instruments in the four-rod fixation system were 810.7,148.0,148.0,134.6 and 14.4 MPa,compared to 655.0,109.9,109.9,119.1 and 11.4 MPa in the MGT.In total sacrectomy model under veaical compression,the range of motion of L3 vertebral body and max van Mise stress of instruments in the four-rod fixation system were 10.9mm and 2172 MPa compared to 49.1 mm and 4325 MPa in the MGT.Conclnsion Comparing with the MGT.the four-rod fixation system was stiffer and showed a significant increase in the stability of lumbosacral area.When total sacrectomy without bone graft were performed,the max stress of the four-rod fixation system was much lower than that of the MGT,which means smaller risk of instrument failure.When subtotal sacrectomy or bone graft were performed,despite better stability,the higher stress shielding-effect of the four-rod fixation svstem might be unfavorable for bone fusion,which still need to be evaluated by clinical trials.

Old distal humeral fractures of type B treated with hinged external fixators in combination with internal fixation / 中华创伤骨科杂志

Objective To evaluate the hinged external flxator in combination with internal fixation for treatment of old type B fractures of the distal humerus.Methods Eleven cases of old type B fractures of distal humerus treated in our department were studied retrospectively.They were treated with hinged external fixators in combination with internal fixation and evaluated according to Mayo elbow performance scores.Results All the patients were followed up for 10 to 18(mean, 12.5)months.The mean union time was 4.6(range, 3.5 to 6.0)months.The final range of flexion to extension of the elbow was 19.4°±6.5° to 124.2°±11.3°.The final ranges of pronation and supination were 72.8°±5.4° and 67.6°± 7.3°.By Mayo elbow performance scores, the post-operative functional outcome of the elbow was excellent in 5 patients, good in 4, and fair in 2.Conclusion The old type B fractures of distal humerus can be effectively treated with hinged external fixators in combination with internal fixation, for early rehabilitation and restoration of the elbow function can be facilitated.

Compound nerve conduit promoting peripheral nerve regeneration in rats / 中华创伤骨科杂志

Objective To explore the role of compound nerve conduit, made of nerve growth factors (NGF) encapsuled by biodegradable core-shell nanofibers through coaxial electrospinning, in regeneration of injured sciatic nerves in rats. Methods The compound nerve conduits were developed from the core-shell structured biodegradable nanofibers with P(LLA-CL) as a shell and BSA/NGF or BSA as a core through coaxial electrospinning. Seventy-two Sprague-Dawley rats were randomly divided into 4 even groups. The middle segments (10 mm) of the sciatic nerve were excised and the defects were repaired with sciatic nerve autograft (group A), with P(LLA-CL) conduit (group B), with PLLA-CL conduit and one injection of NGF (group C), and with P(LLA-CL)/NGF controlled-release conduit (group D), respectively. Morphologic and functional evaluations of nerve regeneration were done by gross observation, sciatic function index, neural electrophysiological examination, resumption rates of triceps weight, histological and ultrastructural observa-tion respectively in 1, 2, 3 months after the operation. Results Three months after the operation, although partial biodegradation and small cracks could be observed, conduits remained intact in outline. Based on the functional and histological observations, nerve regeneration, nerve fibers arrangement, myelination and nerve function reconstruction in the P(LLA-CL)/NGF controlled-release conduit (group D) were similar to those in nerve autograft (group A) and significantly superior to those in groups B and C (P<0.05). Conclusion As the P(LLA-CL)/NGF-controlled release conduit has favorable mechanical properties and biocompatibility, it can effectively promote regeneration of the sciatic nerve in rats.