Objective Lipocalin-2(LCN2) can promote the M1 approach of microglia.The study was to explore the effect of activated microglia induced by LCN 2 on depression pathogenesis in rats and its mechanism . Methods According to the chronic stress depression model created by Willner , 40 adult male SD rats were divided into 4 groups(n=10): normal control group;UCMS group;UCMS+LCN2 siRNA group;LCN2 siRNA control group .A series of stress stimulation was given on UCMS group and UCMS +LCN2 siRNA group for 21 days to create depression model .At 7 days after the stress stimulation , the rats in UCMS+LCN2 siRNA group and LCN2 siRNA control group were anaesthetized by 0.4mg intraperitoneal injection of 10% chloral hydrate , followed by intrathecal injection of LCN2 siRNA(0.015 μL/g, 3 times a week) to the rats till the end of the stress (21 days).At the same time, the same volume of isotonic saline was given to normal control group and UCMS group .The weight of rats was measured every week and the sucrose preference and the forced swimming test were applied to measure behavior of the rats after the experiment .The hippocampus of the rats were extracted and immunofluorescence and western blot were applied to detect the expressions of microglia specific markers:LCN2 and the Iba. Results At 3 week, the weight of rats in UCMS+LCN2siRNA group was higher than that of UCMS group ([262.82 ±0.01]g vs [179.98 ±0.08]g, P<0.05).The weight of rats in normal control group and LCN2 SiRNA control group increased significantly higher than the other two groups (P<0.05).The sucrose preference values of normal control group (0.82 ±0.01),UCMS+LCN2 siRNA group(0.81 ±0.01) and LCN2 siRNA control group(0.82 ±0.01) were higher than that of UCMS group (0.25 ±0.04) (P<0.05).The fixed time of the forced swimming test of UCMS+LCN2siRNA group decreased significantly compared with UCMS group ([4.64 ±0.8]s vs [23.11 ±2.63]s, P<0.05).The LCN2 expression of UCMS group was significantly greater than the other groups (P<0.05).The Iba1 expression in the hippocampus of the UCMS group increased significantly compared with other groups . Conclusion LCN2 is associated with the pathogenesis of de-pression induced by chronic stress reaction and is mechanism may be related to the activation of microglia in the central nervous system of rats.

Objective:To study the immunological characteristics of recombinant Rv3425 protein,to evaluate its diagnosis value and the role in the pathogenicity. Methods: Rv3425 gene was cloned into pET28a vector,the recombinant protein was induced and purified;we analyzed the antigenicity and specificity of Rv3425 by ELISA ( Enzyme-linked immuno sorbent assay) and Western blot methods,apoptosis effect of Rv3425 to macrophage was deteced by FCM ( Flow cytometry method ) . Results: Purified prokaryotic expressed protein Rv3425 was acquired,we found Rv3425 could elicit high level of IFN-γin spleen cells and combine with the serum of iH37Rv (inactivated mycobacterium tuberculosis) immunized mice;the specific IgG and IgM antibodies of TB (Tuberculosis) patients were significantly higher than healthy donors;Rv3425 also could induce the necrosis of macrophage. Conclusion:Our study found that Rv3425 had strong antigenicity and could induce the apoptosis of macrophage,these findings were very important for the research of TB diagnosis and pathogenicity.

Objective:To construct the siRNA expression vector of focal adhesion kinase(FAK) gene and inhibit the expression of FAK gene in tongue cancer cell line Tca8113 by RNA interfering technique. Methods:According to the encoding sequence of FAK mRNA, two pieces of oligonucleotide sequences were designed and synthesized. The annealed oligonucleotide fragments were subcloned into pGCSilencerTM-U6/Neo siRNA expression vector. After being identified by restriction enzyme method, the recombinant pSilencer-FAK plasmids were transfected into Tca8113 cells. The transfected cells were selected by G418 method. Immuocytochemistry and Western blotting were used to evaluate FAK gene silencing efficiency. Results:The oligonucleotide fragments were correctly inserted into pGCSilencerTM-U6/Neo vector. FAK expression of the transfected cells was significantly down-regulated by pSilencer-FAK. Conclusion:The siRNA expression vector of FAK is successfully constructed and FAK expression of Tca8113 cells can be inhibited by RNA interfering technique.

Aim To explore the effect of knockdown spinal cord LCN2 by RNAi on the development of mor-phine tolerance in normal rats.Methods After suc-cessful intrathecal implantation, fourty-eight male Sprague-Dawley rats weighing 1 80 -220 grams were randomly divided into 4 groups (n =1 2):group I:control group,group II:morphine tolerance group, group Ⅲ:mismatch siRNA group,group IV:LCN2 siRNA group.The sixth day after intrathecal implanta-tion,rats were tested to ensure the position of cathe-ters,and it was recorded as d 0.On d 2 -8,rats were subcutaneously (s.c)injected of normal saline (NS) (group I)or morphine (group Ⅱ,Ⅲ,Ⅳ)1 0 μg· g -1 twice a day at 8:00 and 1 6:00.Before everyday s. c injection,rats were intrathecally injected of 1 0 μL DEPC solution (group Ⅰ,Ⅱ),1 0 μL DEPC solution containing 4 μg mismatch siRNA (group III)and 4 μg LCN2 siRNA solution (group IV).Paw withdrawal la-tencies to thermal stimuli (PWTL)were tested before morphine injection and 45 minutes after morphine in-jection on d 1 and d 9.The percentage of maximal pos-sible effect (% MPE)was calculated later.Animals were sacrificed on d 9 after the behavioral test and the lumbar enlargement segments of the spinal cord were removed for detecting the expression of phosphorylated-p38 mitogen-activated protein kinase (p-p38 MAPK) and LCN2 by Western blot and microglia marker Iba1 by immunofluorecence.Results On d 1 ,there was no significant difference in %MPE among four groups. On d 9,compared to group Ⅰ,%MPE was signifi-cantly reduced (P <0.05)while p-p38MAPK,LCN2 and Iba1 were markedly up-regulated in group Ⅱ andⅢ (P <0.05 ).On d 9,compared to group Ⅱ,%MPE was significantly increased while p-p38MAPK, LCN2 and Iba1 were markedly reduced in group IV (P<0.05).Conclusion Using LCN2 siRNA to knock-down spinal LCN2 relieves the development of mor-phine tolerance in normal rats possibly through inhibi-ting the activation of microglia and p38 MAPK in the spinal cord.

OBJECTIVETo investigate the expression of full-length spleen tyrosine kinase [SYK (L)] mRNA and protein in human oral squamous cell carcinoma (OSCC) as well as its possible effects on the invasion and metastasis of OSCC.

METHODSThe expression of SYK (L) was detected in 27 cases of OSCC tissues and its matched adjacent non-cancerous tissues by real-time quantitative polymerase chain reaction (RT-qPCR), Western blot, and immunohistochemistry. Fourteen cases of normal oral gingival tissues were also analyzed as a normal control.

RESULTSReduced mRNA and protein expression of SYK (L) in OSCC tissues was observed compared with that in normal oral gingival tissues (P<0.01) and adjacent non-cancerous tissues (P<0.05). SYK(L) expression was significantly associated with lymph-node metastasis (P<0.05).

CONCLUSIONSYK(L) is a candidate tumor suppressor for OSCC tissues, and has an inhibitive effect on the initiation, proliferation, and lymph-node metastasis of human OSCC.

Blotting, Western ; Carcinoma, Squamous Cell ; metabolism ; Humans ; Immunohistochemistry ; Lymphatic Metastasis ; Mouth Neoplasms ; metabolism ; RNA, Messenger ; Syk Kinase ; metabolism

Objective To explore the application effect of modified installing method of high flexion rotating platform prosthesis trial model in double compartment knee arthroplasty.Methods A total of 20 patients(26 knees)with severe knee osteoarthritis admitted to Hebi People's Hospital from January 2021 to April 2022 and required double compartment knee arthroplasty were selected as the research subjects.Patients were divided into the control group(10 knees)and the observation group(16 knees)based on the installing method of high flexion rotating platform prosthesis trial model.The patients in the control group used the conventional installing method of high flexion rotating platform prosthesis test model(placing the tibial side test model,the femoral side test model and finally the pad test model),while patients in the observation group used the modified installing method of high flexion rotating platform prosthesis test model(matching the femoral test model,pad and tibial test model when the patient flexed 60° to 90°,and placing them together into the knee joint osteotomy groove).The intraoperative bleeding volume and surgical time of patients in the two groups were recorded.Before surgery,2 weeks after surgery and 18 months after surgery,the knee joint function of patients was evaluated by the American hospital for special surgery(HSS)scoring system,and the range of motion(ROM)of knee joint of patients was measured by protractor.Results The intraopera-tive bleeding volume of patients in the observation group was significantly less than that in the control group,and the surgical time was significantly shorter than that in the control group(P＜0.05).There was no statistically significant difference in preoperative HSS score and ROM of knee joint of patients between the two groups(P＞0.05).At 2 weeks and 18 months after surgery,the HSS score and ROM of knee joint of patients in the observation group were significantly higher than those in the control group(P＜0.05).Conclusion Compared with the conventional installing method of high flexion rotating platform prosthesis trial method,the modified installing method of high flexion rotating platform prosthesis trial model in double compart-ment knee arthroplasty for patients with severe knee osteoarthritis can effectively improve surgical efficiency,reduce intraopera-tive bleeding,improve knee function and increase ROM of knee joint.

Objective To observe the effect of Shoutai Pills on endometrial decidualization of mice with recurrent spontaneous abortion(RSA);To explore its possible mechanism in the treatment of RSA based on histone modification.Methods Totally 40 female CBA/J mice were divided into normal group,model group,Shoutai Pills low-dosage group(7.5 g/kg),Shoutai Pills high-dosage group(15 g/kg)and dydrogesterone group(3 mg/kg).The normal group were co housed with BALB/C male mice,while the other groups were co housed with DBA/2 male mice to establish an RSA mouse model.After modeling,the administration groups were given corresponding medication solution by gavage,while the normal group and model group were given equal volume of pure water by gavage for 10 consecutive days.The embryo condition was observed and the embryo loss rate was calculated,ELISA was used to detect serum prolactin(PRL)content,HE staining was used to observe the morphological changes of decidual tissue,RT-PCR was used to detect PRL mRNA expression in decidual tissue,Western blot was used to detect the protein expressions of H4ac,H3K27ac,H3K27me3.Results Compared with the normal group,the model group mice showed a significant increase in embryo loss rate,a significant decrease in serum PRL content,disordered arrangement of decidual cells,and extensive bleeding and necrosis;the expression of PRL mRNA and protein in decidual tissue significantly decreased,the protein expressions of H4ac and H3K27ac significantly decreased,while the expression of H3K27me3 protein significantly increased,with statistical significance(P<0.05).Compared with the model group,the embryo loss rate of Shoutai Pills low-and high-dosage groups and the dexamethasone group significantly decreased,the serum PRL content significantly increased,tightly arranged decidual cells,reduced necrosis,and intact glands;the expression of PRL mRNA and protein in decidual tissue of mice in Shoutai Pills high-dosage group and the dexamethasone group significantly increased,the protein expressions of H4ac and H3K27ac significantly increased,the expression of H3K27me3 protein significantly decreased,with statistical significance(P<0.05).Conclusion Shoutai Pills can promote endometrial decidualization in RSA mice,which is related to the changes of histone modification in endometrial stromal cells.

Objective:To explore the clinical efficacy of double traction-assisted reduction internal fixation and open reduction internal fixation in treating tibial plateau fractures.Methods:Data of patients with tibial plateau fracture admitted to West China Hospital of Sichuan University from January 2016 to December 2021 were retrospectively analyzed, and patients were divided into two groups according to treatment method: double traction-closed reduction internal fixation group (referred to as double traction group) and open reduction internal fixation group (referred to as open group). The double traction group included 21 patients, with 15 male and 6 female patients, with a mean age of 56.14±9.24 years (range, 45-72 years). Schatzker classification of fractures: 1 type I, 2 type II, 2 type III, 5 type IV, 6 type V, and 5 type VI. The open group included 29 patients, with 20 male and 9 female patients, with a mean age of 58.97±4.84 years (range, 47-70 years). Schatzker classification of fractures: 2 type I, 4 type II, 8 type III, 4 type IV, 5 type V, and 6 type VI. The surgical time, incision length, intraoperative blood loss, length of hospital stays, fracture healing time, postoperative time to full weight bearing, Rasmussen score, Hospital for Special Surgery (HSS) knee score, and complications were compared between the two groups of patients.Results:Both groups were followed up for 24 to 36 months, with an average of 30 months. There were significant differences in the operation time (92.61±6.22 min vs. 47.92±9.53 min), incision length (4.54±0.56 cm vs. 6.26±0.51 cm), and intraoperative blood loss (47.05±9.72 ml vs. 156.82±4.62 ml) between the group treated with closed reduction and double traction and the group treated with open reduction, with statistical significance ( t=18.83, 10.78, 53.24, P<0.001). There were also significant differences in the hospitalization time (5.35±0.41 d vs. 5.84±0.78 d), fracture healing time (3.72±0.74 months vs. 4.22±0.42 months), and time to full weight-bearing after surgery (11.29±1.10 weeks vs. 15.07±1.96 weeks) between the two groups, with statistical significance ( t=2.30, P=0.026; t=3.38, P<0.001; t=7.96, P<0.001). The HSS score at 6 months after surgery in the group treated with closed reduction and double traction was 81.61±2.32 points, which was higher than the score in the group treated with open reduction (77.66±4.01 points), with statistical significance ( t=4.07, P<0.001); at 12 months after surgery, the Rasmussen score in the group treated with closed reduction and double traction was 16.71±1.00 points, which was higher than the score in the group treated with open reduction (13.79±1.42 points), with statistical significance ( t=8.05, P<0.001). There was no fracture malunion or compartment syndrome occurred in both groups. The incidence of complications was 5% (1/21) in the group treated with closed reduction and double traction, and 10% (3/29) in the group treated with open reduction, with statistical significance (χ 2=0.52, P=0.473). Conclusion:The advantages of double traction-assisted reduction and internal fixation for tibial plateau fractures include minimal trauma, minimal bleeding, early mobilization, and shorter fracture healing time. It is a safe and reliable treatment method.

ObjectiveTo investigate the therapeutic effect of Yupingfeng San on allergic rhinitis （AR） and its effect on Reactive oxygen species （ROS）/NOD-like receptor thermal protein domain associated protein 3 （NLRP3）/cysteinyl aspartate specific proteinase-1 （Caspase-1） pathway. MethodSPF mice were randomly divided into control group， model group， loratadine group （0.9 mg·kg^-1）， and low， medium， and high dose Yupingfeng San groups （6， 12， 24 mg·kg^-1）， with 10 mice in each group. The control group was given routine feeding， and the other groups were intraperitoneally injected with ［ovalbumin（OVA） + Al（OH）₃ + phosphate buffer solution（PBS）］ suspension once every other day for seven consecutive times. After seven days， 10% OVA solution was instilled in the nose， two times each day for seven consecutive days. After successful modeling， each administration group was intraperitoneally injected with different doses of Yupingfeng San Decoction， and the control group and model group were intraperitoneally injected with an equal volume of normal saline. Symptoms of sneezing， scratching， and runny nose were recorded and scored daily. The levels of ovalbumin specific immunoglobulin E （OVA-sIgE）， histamine， eosinophil cationic protein （ECP）， prostaglandin D₂ （PGD₂）， interleukin-1β （IL-1β）， interleukin-18 （IL-18）， interleukin-4 （IL-4）， and γ interferon （IFN-γ） in nasal lavage solution and serum of mice were detected by enzyme-linked immunosorbent assay （ELISA）. The damage status of the nasal mucosa was observed by hematoxylin-eosin （HE） staining. The number of goblet cells in the nasal mucosa of mice was observed by periodic acid-Schiff （PAS） staining. The expression of NLRP3 protein in the nasal mucosa of mice was detected by immunohistochemistry. Western blot was used to detect the expressions of NLRP3， cleaved Caspase-1， and cleaved gasdermin D （GSDMD） proteins in the nasal mucosa. The test kit was used to detect the changes in ROS in the nasal cavity of mice in each group. ResultCompared with the control group， the nasal symptoms of the model group were significantly aggravated， and the levels of inflammatory factors OVA-sIgE， histamine， ECP， PGD₂， IL-1β， IL-18， and IL-4 in serum and nasal lavage solution were increased （P<0.05，P<0.01）. The levels of IFN-γ were decreased （P<0.05，P<0.01）. The histopathological score， goblet cell number， and ROS content were significantly increased （P<0.01）， and the expressions of pyrodeath-related proteins NLRP3， cleaved Caspase-1， and cleaved GSDMD were increased （P<0.01）. Compared with the model group， the nasal symptoms of the loratadine group and Yupingfeng San groups were significantly relieved， and the levels of inflammatory factors OVA-sIgE， histamine， ECP， PGD₂， IL-1β， IL-18， and IL-4 in serum and nasal lavage solution were decreased （P<0.05，P<0.01）. The levels of IFN-γ were increased （P<0.05，P<0.01）. The histopathological scores， goblet cell number， and ROS content were significantly decreased （P<0.05，P<0.01）， and the expressions of pyrodeath-related proteins NLRP3， cleaved Caspase-1， and cleaved GSDMD were decreased （P<0.05，P<0.01）. Compared with the loratadine group， the curative effect of the high dose Yupingfeng San group was further increased （P<0.05，P<0.01）. ConclusionYupingfeng San has a therapeutic effect on AR， and its specific effect may be related to the inhibition of ROS/NLRP3/Caspase-1-induced cell pyroptosis.

At the present day, curettage and periodontal surgery comprise the main strategy for the treatment of periodontitis, however, these methods are limited in regenerating cementum. It has been found that some biological factors such asenamel matrix derivative (EMD), transforming growth factor-β (TGF-β) and insulin-like growth factor (IGF) could promote cementum regeneration. In the cementum regenerationstudies, there has been a lack of criteria to distinguish cementum from alveolar bone and other types of cementum. Therefore, this article will briefly review the biological factors that affect the cementum regeneration and the molecular markers used to judge the regenerating cementum.