Objective To observe the change of JAK-STAT-SOCS signal molecules after interferons alpha acting on the cancerous oral cells in 3 different degrees,namely NOK,DOK and KB cells,and to provide research foundation for the deep understanding of OSCC (oral squamous cell cancer) tumor ceils immune escape mechanism.Methods NOK,DOK,and KB cells were all cultured respectively,and then the third passage cells in the logarithmic growth phase were inoculated in cell culture plate.Blank control group of each hole was added 2 ml complete medium containing 10 ％ FPS.DMSO control group of each hole was added 2 ml complete medium containing 0.1％ DMSO.And in experimental groups containing 10 U/ml,100 U/ml,and 500 U/ml interferons,complete culture medium were added to each hole containing different concentrations of interferons alpha.CP-690550 (100 μmol/L) was added before interferons alpha was added 1 h.All were detected by RT-PCR test and Western blot test after conventional cultured for 24 h.Results RT-PCR detection showed that JAK1 and JAK2 in NOK cells had a small amount of expression,interferons alpha and CP-690550 cells could not influence the expression of JAK1 and JAK2 of NOK group,and there was no statistically significant difference (P ＞ 0.05).Interferons alpha in 100 and 500 U/ml could stimulate the increase of JAK1 and JAK2 expression in DOK and KB cells,and the differences were statistically significant (P ＜ 0.05).CP-690550 could effectively reduce the JAK1 expression of DOK and KB cells,while had no effect on the expression of JAK2,and the differences were statistically significant (P ＜ 0.05).Western blot showed that STAT1,STAT3 and pSTAT3 (Tyr705) all expressed in the control group,while pSTAT1 (Tyr701) didn't express in the control group.Interferons alpha and CP-690550 cells had no effect on STAT1,STAT3 and pSTAT3 (Tyr705) expression of NOK group,and there was no statistically significant difference (P ＞ 0.05).100 U/ml and 500 U/ml of interferons alpha could stimulate the increases of pSTAT3 (Tyr705) expression of DOK and KB cells,and the differences were statistically significant (P ＜ 0.05).While they had no effect on pSTAT1 (Tyr701) expression.CP-690550 could effectively reduce the pSTAT3 DOK and KB cells (Tyr705) expression,and the differences were statistically significant (P ＜ 0.05).Western blot showed that there were expression of SOCS1 and SOCS3 in control group.Interferons alpha and CP-690550 had no effect on SOCS1 and SOCS3 expression of NOK cell group,and there was no statistically significant difference (P ＞ 0.05).100 U/ml and 500 U/ml of interferons alpha could stimulate the increase of SOCS 1 expression of DOK and KB cells,and differences were statistically significant (P ＜ 0.05).For the expression of SOCS3,no influence.CP-690550 could effectively reduce the expression of SOCS1 of DOK and KB cells,and the differences were statistically significant (P ＜ 0.05).Conclusions Interferons alpha activate DOK JAK1 and KB cells and the expression of JAK2,mainly JAK1 activation.Interferons alpha,by activating DOK JAK1 and KB cells and the expression of JAK2,promote STAT3 phosphorylation in Tyr705 locus.Interferons alpha,by promoting STAT3 phosphorylation,further promote the expression of SOCS1,which plays the role in inhibiting interferons alpha and reducing the apoptosis.

Objective To explore the effect of expression of protein kinase C receptor 1(RACK1) induced by lipopolysaccharide (LPS) on Sonic hedgehog(SHH) signaling pathway in rat puhnonary microvascular endothelial cells (RPMVEC).Methods The healthy male SPF grade SD rat with 100-120 g body weight were gotten from the laboratory animal center of Anhui province.Using immunocytochemistry method,the expression of RACK1 protein in RPMVECs was detected,cultured RPMVECs were randomly divided into different groups as LPS dose-dependent group,SAG(smoothened Agonist,a SHH signaling pathway specific agonist) dose-dependent group,LPS time-dependent group,SAG time-dependent group and LPS+SAG group.In LPS dose-dependent groups,RPMVECs were cultured with 0.1,1,10 mg/L LPS for 8 h.In LPS time-dependent groups,RPMVECs were cultured with 10 mg/L LPS for 0,2,4,8,12,24 h.In SAG dose-dependent groups,RPMVECs were cultured with 0.1,1,10 μ mol/L for 8 h.In SAG time-dependent groups,RPMVECs were cultured with 1 μ mol/L SAG for 0,2,4,8,12,24 h.In LPS+SAG group,RPMVECs were cultured with 1 μ mol/L SAG 8 h after 10 mg/L LPS treatment for 1 h.In addition,blank group,LPS group and SAG group were set for control.Western blot were used to detect the level of RACK1 and RT-PCR were used to detect the expression of GLI-1 mRNA after intervention.Results Immunocytochemistry revealed that RACK1 were present in RPMVEC.1.In LPS dose-dependent groups (0,0.1,1,10 mg/L),the level of RACK1 elevated as LPS dose increased correspondingly with inter-group difference (P＜0.05);the relative expression levels of GLI-1 mRNA were (1.109 + 0.063),(1.039 + 0.135),(0.813 ± 0.066),(0.770 + 0.105),(1 mg/L vs.10 mg/L,P＞0.05;the rest P＜0.05).In LPS time-dependent groups,the relative expression level of RACK1 at 2 h (0.370 + 0.010) was higher than that at 0 h (0.329 ± 0.008),peaked at 12 h (1.296 ± 0.048),and compared with 0 h,there was significant differences (F=1 272.204,P＜0.05).The relative expression level of GLI-1 mRNA was decreased at 2 h (0.929 ±-0.007),and compared with 0 h(1.089 ± 0.042),there was significant differences (F=306.609,P＜0.05).2.In SAG dose-dependent groups,there was no significant difference in level of RACK1 between groups(all P＞0.05).The relative expression levels ofGLI-1 mRNA were (1.109 ± 0.063),(1.169 ± 0.052),(3.468 ± 0.128),(3.434 ± 0.054),(0 μ mol/L vs.0.1 μ mol/L and 1 μ mol/L vs.10 μ mol/L,P＞0.05,the rest P＜0.05).Among SAG time-dependent groups,there was no significant difference in levels of RACK1 protein(P＞0.05).The relative expression level of GLI-1 mRNA increased at 2 h (3.027 ± 0.065),and compared with 0 h (2.651 + 0.123),there was significant differences (F=132.841,P＜0.05).3.In LPS+SAG intervention groups,the expression of RACKI was lower than that in LPS group (0.831 ±0.040 vs.1.189 ± 0.149,P＜0.05),and the expression of GLI-1 mRNA was higher than that in LPS group (2.720 + 0.130 vs.0.796-4-0.082,P＜0.05).Conclusions The LPS up-regulates the expression of RACK1 in RPMVECs,and the activated SHH signaling pathway can down-regulate the expression of RACK1 induced by LPS in RPMVECs.

Objective:To explore the clinical effect of antibiotic bone cement combined with delayed lateral supramolleolar perforator fascial flap in the treatment of diabetic foot(DF).Methods:From April 2020 to July 2021, a total of 6 patients with DF were treated with antibiotic bone cement combined with delayed lateral supramolleolar perforator fascial flap. The patients were 5 males and 1 female, aged from 45 to 67 years old with an average of 56.2 years old. The wounds were all located in dorsal foot, 4 in right foot and 2 in the left. The wound area was 2.4 cm×5.0 cm-6.5 cm×10.0 cm. The depth of wound were: 3 cases up to tendon layer, and 3 cases up to metatarsal bone. Two of the wound were complicated with metatarsal osteomyelitis. The wounds at Wagner grade 3 in 4 patients and grade 4 in 2 patients. The flap size was 3.0 cm×6.0 cm-8.0 cm×11.0 cm. All of the wounds were repaired with delayed supramolleolar perforator fascia flap after debridement, application of antibiotic bone cement and fumigation with Sanhuang decoction(a traditional Chinese medicine). The affected limbs were externally fixed with plaster and raised after surgery, and the colour, temperature, tension and capillary reaction of the flaps were closely observed. Stitches were removed 2 weeks after surgery and rehabilitation of the affected limb was performed. Regular follow-up was made postoperatively. The appearance of flaps and the scar of donor and recipient sites were observed. The foot and ankle function were evaluated by the American Orthopaedic Association foot and Ankle Surgery(AOFAS) score scale.Results:Six cases of DF had no recurrence of wound infection. All flaps survived well. The average follow-up time was 6(3-14) months. The postoperative follow-up revealed satisfactory appearance of the flap, only linear scars remained in the donor and recipient sites. The function of foot and ankle recovered well with full weight-bearing and normal walk. AOFAS scores ranged from 81 to 95.Conclusion:It is an effective method to treat DF by applying antibiotic bone cement combined with delayed superior lateral malleolus perforator fascial flap. The operation is simple, safe and can cut down the time of treatment, quickly control the wound infection. It deserves further trials.