Objective To analyze the expression and clinical significance of stimulator of interfer-on genes(STING),C-C motif chemokine ligand 5(CCL5),interferon regulatory factor 3(IRF3)and programmed death ligand-1(PDL1)in squamous cell lung cancer.Methods A total of 56 pa-tients with squamous cell lung cancer were enrolled.Resected tumor tissues and adjacent non-tumor tissues(located more than 5 cm from the tumor margin)were collected.Immunohistochemical staining was performed to detect the expression of STING,CCL5,IRF3 and PDL1.The correlations of STING,CCL5,IRF3 and PDL1 with clinical data were analyzed.The relationship between the expression of STING,CCL5,IRF3 and PDL1 in lung squamous cell carcinoma tissues and prognosis was also evalua-ted.The prognostic factors of patients with lung squamous cell carcinoma were analyzed.Results The positive rate of STING expression in lung squamous cell carcinoma tissues was significantly lower than that in adjacent non-tumor tissues,whereas the positive rates of CCL5,IRF3 and PDL1 were significantly higher(P＜0.05).The expression levels of STING,CCL5,IRF3 and PDL1 were associated with tumor diameter,TNM stage,lymph node metastasis and differentiation degree(P＜0.05).The 3-year survival rate of STING positive expression patients was significantly higher than that of STING negative expression patients(P＜0.05).The 3-year survival rate of CCL5 positive,IRF3 positive and PDL1 positive expression patients was significantly lower than that of CCL5 negative,IRF3 negative and PDL1 negative expression patients(P＜0.05).STING,CCL5,IRF3 and PDL1 were identified as prognostic factors for patients with squamous cell lung cancer(P＜0.05).Conclusion In squamous cell lung cancer tissues,STING is expressed at low levels,while CCL5,IRF3 and PDL1 are ex-pressed at high levels.These findings have significant clinical value in assessing the prognosis of pa-tients with squamous cell lung cancer.

Objective To investigate the effect of ankyrin-repeat domain-containing protein 22(ANKRD22)on the proliferation,invasion,and migration of human hepatoma cells and its molecular mechanism.Methods The TCGA database was used to analyze the expression level of ANKRD22 in normal liver tissue and hepatocellular carcinoma tissue and its association with prognosis.Western Blot and qRT-PCR were used to measure the expression of ANKRD22 in human normal liver cells(L-02)and human hepatoma cells(Huh7,HepG2,MHCC-97H,SK-HEP-1,and SMMC-7721);CCK-8 assay,EdU,wound healing assay,and Transwell assay were used to observe the effect of ANKRD22 on the proliferation,invasion,and migration of hepatoma cells;Western Blot was used to investigate the association of ANKRD22 with cyclins and EMT-related proteins;KEGG and ssGSEA analyses were performed to investigate the mechanism of action of ANKRD22 in hepatoma cells,and related experiments were conducted for validation.The independent-samples t-test was used for comparison of continuous data between two groups;a one-way analysis of variance was used for comparison between multiple groups,and the least significant difference t-test was used for further comparison between two groups.Results In the TCGA database,the expression level of ANKRD22 in hepatoma tissue was significantly higher than that in normal liver tissue(t=5.083,P<0.05),and the patients with a high expression level of ANKRD22 had longer overall survival and disease-related survival than those with a low expression level of ANKRD22(P<0.05).The expression level of ANKRD22 in various human hepatoma cell lines was higher than that in human normal liver cells(all P<0.05).Cell proliferation assay showed that the ANKRD22 overexpression group had significantly higher EdU positive rate and proliferation rate than the Vector group(t=19.60 and 6.72,both P<0.001),and compared with the si-NC group,the si-ANKRD22#2 group and the si-ANKRD22#3 group had significantly lower EdU positive rate and proliferation rate(all P<0.001).Compared with the Vector group,the overexpression group had significantly higher expression levels of Cyclin E1,Cyclin D1,CDK7,and CDK4(t=3.54,4.95,6.34,and 5.19,all P<0.01),and the si-ANKRD22#2 group and the si-ANKRD22#3 group had significantly lower expression levels than the si-NC group(all P<0.001).The overexpression group had a significantly lower expression level of P27 than the Vector group(t=6.12,P<0.001),and the si-ANKRD22#2 group and the si-ANKRD22#3 group had a significantly higher expression level than the si-NC group(both P<0.001).Invasion and migration experiments showed that compared with the Vector group,the ANKRD22 overexpression group had significantly higher migration rate and number of crossings through the membrane(migration group and invasion group)(t=5.01,25.60,and 3.67,all P<0.05),and compared with the si-NC group,thesi-ANKRD22#2 group and the si-ANKRD22#3 group had significantly lower migration rate and number of crossings through the membrane(migration group and invasion group)(all P<0.01).The overexpression group had significantly higher expression levels of N-cadherin,Vimentin,and Snail than the Vector group(t=12.13,8.85,and 13.97,all P<0.001),and the si-ANKRD22#2 group and the si-ANKRD22#3 group had significantly lower expression levels than the si-NC group(all P<0.001);the overexpression group had a significantly lower expression level of E-cadherin than the Vector group(t=4.98,P<0.01),and the si-ANKRD22#2 group and the si-ANKRD22#3 group had a significantly higher expression level than the si-NC group(both P<0.001).The KEGG enrichment analysis and the ssGSEA analysis showed that ANKRD22 was associated with the PI3K/AKT/mTOR signaling pathway in hepatocellular carcinoma,and the overexpression group had significantly higher expression levels of p-AKT/AKT,p-PI3K/PI3K,and p-mTOR/mTOR than the Vector group(t=12.21,3.43,and 9.75,all P<0.01),and the si-ANKRD22#2 group and the si-ANKRD22#3 group had significantly lower expression levels than the si-NC group(all P<0.001).Conclusion ANKRD22 is highly expressed in hepatoma cells and can promote the proliferation,invasion,and migration of hepatoma cells and the activation of the PI3K/AKT/mTOR signaling pathway.

Objective:To compare the mid-term clinical effects of AngioJet rheolytic thrombectomy assisted catheter-directed thrombolysis (ART+CDT) with catheter-directed thrombolysis (CDT) in the treatment of acute deep venous thrombosis of lower extremities.Methods:Ninety-one patients admitted to the Department from Jan 2016 to Dec 2017 were placed with inferior vena cava filters and divided into ART+CDT group (30 cases)and CDT group (61 cases). Total urokinase dosge, thrombolytic time, operative cost, length of hospital stay, detumescence rate, thrombus clearance rate, cumulative patency rate of lower limb veins, Villalta score at 2 years and 5 years, thrombosis recurrence rate and chronic venous insufficiency quality of life questionnaire were compared between the two groups.Results:The success rate of surgery was 100% in both groups, there was no mortality. There were significant differences in the short-term postoperative outcomes between the two groups in terms of total dosage of urokinase, thrombolysis time, total cost of surgery, length of hospital stay, detumescence rate, venous patency scores before and after treatment, and venous patency rate (all P<0.05). For the mid- and long-term postoperative outcomes of 2 and 5 years, there were no significant differences in the incidence of PTS, recurrence rate of thrombus, chronic venous function scale, and cumulative patency rate at 2 years (all P>0.05). Conclusions:ART+CDT has a significant advantage over CDT alone in terms of early efficacy and early reopening of blood flow in patients. Both ART+CDT and CDT have a low incidence of PTS and a low recurrence rate of thrombus in the mid-term follow-up, and both have satisfactory performance in the mid- and long-term efficacy of interventional treatment of deep venous thrombosis of lower limbs.

Objective To explore clinical effect of Jiawei Sangju decoction combined with oseltamivir in the treatment of influenza B complicated with benign acute childhood myositis(BACM).Methods Sixty children with influenza B complicated with BACM hospitalized in the Department of Pediatrics,Wenzhou Hospital of Integrated Traditional Chinese and Western Medicine from January 2021 to January 2024 were selected and divided into control group and study group according to random number table method,with 30 cases in each group.The control group was treated with oseltamivir,and the study group was treated with Jiawei Sangju decoction combined with oseltamivir.The clinical efficacy,symptom improvement time and aspartate transaminase(AST),alanine transaminase(ALT),creatine kinase(CK),creatine kinase isozyme(CK-MB),lactate dehydrogenase(LDH)and myoglobin(Mb)before and after treatment were compared between two groups.Results After treatment,the total effective rate of study group was significantly higher than that of control group(χ2=5.455,P=0.020).The duration of fever and leg pain relief in study group were significantly shorter than those in control group(P<0.05).After 5 days of treatment,the levels of AST,CK,CK-MB,LDH and Mb in two groups were significantly lower than before treatment(P<0.05),and the levels of AST,CK,CK-MB,LDH and Mb in study group were significantly lower than those in control group(P<0.05).Conclusion Jiawei Sangju decoction combined with oseltamivir can improve the symptoms of influenza B with BACM,reduce the levels of muscle enzyme and Mb,and have a positive effect on the prognosis of children.

Objective:To investigate the clinical effect of using free bilateral anterolateral thigh flaps(ALTF) in series to repair large area soft tissue defects of forearm.Methods:The clinical data of 11 patients with large soft tissue defects of forearm admitted in the Department of Plastic(Repair and Reconstruction) Surgery, Lishui Hospital of Zhejiang University from March 2014 to December 2021 were retrospectively analyzed, including 8 males and 3 females. Aged 36 to 68 years old, with an average of 48 years old. VSD treatment was performed after debridement, and until fresh removed 3 to 5 days after the operation. Until the wound was fresh. The wound was repaired with free bilateral ALTF in series until fresh. The size of the forearm wound was 18 cm×15 cm-28 cm×13 cm. The cut area of a single flap was 10 cm×8 cm-20 cm×13 cm. The series of bilateral flaps: One of the flaps was used as the proximal flap, and its vascular pedicle was anastomosed with the arteries and veins of the recipient area. The other flap was used as the distal flap, and the arteries and veins between the 2 ALTFs on both sides were anastomosed. The vascular pedicle beyond the distal flap was ligated or anastomosed to the distal end of the ulnar artery or the distal end of the radial artery. The flap and the surrounding skin of the recipient area were sutured immediately. The donor sites of the flap was closed directly. Periodic and regular outpatient follow-up was performed after operation and the clinical efficacy was analyzed.Results:All the flaps successfully survived after the surgery. The postoperative follow-up lasted for 6-18 months, 12 months in average. The flaps survived well with good soft texture, without swelling, the capillary reaction time was normal, without surface ulceration, in rosy colour and restored protective sensations. The hand function of the affected limb recovered well. The wound at donor sites healed well without complications. At the last follow-up, the hand function of the affected limb was evaluated by the Evaluation Trial Standards of Upper Limb Partial Functional of Hand Surgery of Chinese Medical Association, the result was 7 in excellent and 4 in good. The patients were satisfied with the flap and the therapeutic effect.Conclusion:The free bilateral ALTF in series can be used to repair a wound surface with large area, and the donor site can be closed at the same time. It is an effective method to repair large soft tissue defect of forearm.

Bacitracin is a broad-spectrum antibiotics mainly produced by Bacillus, and is used as veterinary medicine in the fields of livestock and poultry breeding. Insufficient supply of precursor amino acids might be an important factor that hinders high-level microbial production of bacitracin. We investigated the effect of strengthening L-cysteine supply on bacitracin production by an industrial bacitracin producer, Bacillus licheniformis DW2. Overexpression of cysK encoding L-cysteine synthase led to a 9.17% increase of the bacitracin titer. Moreover, overexpression of cysE encoding L-serine acetyltransferase and cysP encoding thiosulfate/sulfate intracellular transporter increased the bacitracin titers by 7.23% and 8.52%, respectively. Moreover, overexpression of a putative cystine importer TcyP led to a 29.19% increase of intracellular L-cysteine, and bacitracin titer was increased by 7.79%. Subsequently, the strong promoter PbacA was used to replace the promoters of genes cysP, cysE and tcyP in strain DW2::ysK, respectively. The resulted strain CYS4 (DW2::cysK-PbacA-(cysP)-PbacA(cysE)- PbacA(tcyP) produced 910.02 U/mL bacitracin, which was 21.10% higher than that of the original strain DW2 (747.71 U/mL). Together with the experiments in 3 L fermenters, this research demonstrated that enhancing intracellular L-cysteine supply is an effective strategy to increase bacitracin production of B. licheniformis.
Amino Acids ; Bacillus licheniformis/genetics* ; Bacitracin ; Cysteine ; Metabolic Engineering

Objective:To investigate the relationship between serum fibroblast growth factor 21/23 level, trauma severity and prognosis in middle-aged and older adult patients with traumatic fracture.Methods:A total of 126 middle-aged and older adult patients with traumatic facture who received treatment in the Second People's Hospital of Lishui, China between June 2017 and June 2019 were included in the study group. Fifty healthy controls who concurrently received physical examination in the Second People's Hospital of Lishui were included in the control group. The study group was divided into five subgroups according to relevant criteria: mild, moderate, severe, poor prognosis and good prognosis. The levels of C-reactive protein (CRP), procalcitonin (PCT), FGF21 and FGF23 were measured.Results:On admission, serum CRP, PCT, FGF23, FGF2 levels in the study group were (19.18 ± 5.66) mg/L, (0.71 ± 0.20) μg/L, (79.75 ± 18.62)μg/L,(52.10 ± 16.34) μg/L, respectively, and they were significantly higher than those in the control group [ (7.60 ± 2.61) mg/L, (0.30 ± 0.11) μg/L, (40.18 ± 10.33) μg/L, (30.11 ± 10.19) μg/L, t = 18.888, 17.750, 18.336, 11.032, all P < 0.001). On admission, serum CRP, PCT, FGF23, FGF2 levels in the study group were (19.18 ± 5.66) mg/L, (0.71 ± 0.20) μg/L, (79.75 ± 18.62) μg/L, (52.10 ± 16.34) μg/L, respectively, and they were significantly increased at 1 day [(21.59 ± 4.53) mg/L, (0.79 ± 0.22) μg/L, (83.85 ± 19.07) μg/L, (55.18 ± 16.55) μg/L, t = 3.72, 3.29, 1.56, 1.56, P < 0.05, P < 0.05, P = 0.122, P = 0.122] and 3 days after surgery [(23.15 ± 3.16) mg/L, (0.80 ± 0.24) μg/L, (88.11 ± 19.80) μg/L, (59.70 ± 16.07) μg/L, t = 6.65, 3.12, 3.59, 3.77, all P < 0.05] , and significantly decreased at 7 days after surgery [(14.35 ± 4.02) mg/L, (0.52 ± 0.16) μg/L, (50.06 ± 15.50) μg/L, (32.18 ± 12.52) μg/L, t = 8.31, 8.58, 13.77, 11.11, all P < 0.001]. On admission, there were significant differences in serum CRP, PCT, FGF23, FGF21 levels between mild, moderate and severe groups ( F = 25.087, 15.851, 15.831 and 12.645, all P < 0.001). On admission, serum CRP, PCT, FGF23, FGF21 levels in the poor prognosis group were significantly higher than those in the good prognosis group ( t = 5.757, 4.984, 3.189 and 4.006, all P < 0.001). The receiver operating characteristic curve (ROC) analysis results showed that serum CRP, PCT, FGF23, FGF21 levels in patients with traumatic fracture on admission had a certain value in the prediction of poor prognosis. Combined detection of these four indexes had the highest value, with AUC (0.95 CI) of 0.877 (0.783-0.982). Conclusion:Serum FGF21 and FGF23 levels have a certain value in the prediction of severity and prognosis of traumatic fracture in middle-aged and older adult patients.

Objective:To compare the effect of stripped perforator pedicle on the survival of perforator flap.Methods:From January, 2015 to December, 2019, 44 patients with soft tissue defects of distal shank and ankle were repaired with perforating vessel pedicled propeller flap. According to the nudity of the perforator pedicle, the patients were divided into 2 groups: stripped group ( n=14) and non-stripped group ( n=30). The gender, age, history of smoking, history of diabetes, location of wound, size of flap, perforator artery of flap, closure method of donor site, degree of flap swelling 3 days after operation, percentage of survival area of flap 7 days after operation and postoperative complications were analysed retrospectively. Data were analyzed statistically. The difference was statistically significant when P<0.05. Results:There was no significant difference between the 2 groups in age, smoking history, diabetes history, wound location, size of flap, perforator artery and donor site closure( P<0.05). The degree of swelling of flap in the stripped group 3 days after operation [(+) 94.00%, (++) 6.00%, (+++) 0.00%, (++++) 0.00%)] was less than that in the non-stripped group [(+)47.00%, (++) 29.00%, (+++) 13.00%, (++++) 11.00%)] . The difference was statistically significant between the 2 groups( P<0.05). The percentage of flap survival area in the stripped group [(100.00±0.00) %] was higher than that in the non-stripped group [(88.23±21.29)%] , and the difference was statistically significant ( P<0.05). The incidence of postoperative complications in the stripped group(0.00%) was lower than that in the non-stripped group (39.00%). The difference heel statistically significant( P<0.05). Conclusion:The stripped pedicle of the perforating vessel can promote a better survival of the arterial perforating branch propeller flap, and the degree of postoperative swelling and complications of the flap are lower.

Objective:To investigate the clinical effect of lateral rectus abdominis approach combined with presacral decompression for surgical treatment of old Denis type II sacral fractures complicated with upper sacral plexus injury.Methods:A retrospective case series study was performed on the clinical data of 9 patients with old Denis type II sacral fractures complicated with upper sacral plexus injury (L 4-S 1) admitted to Zhongnan Hospital of Wuhan University from June 2010 to December 2016. There were 6 males and 3 females, aged (33.1±7.5)years (range, 15-58 years). Embolization of internal iliac artery and preimplantation of abdominal aortic balloon were performed 2 hours before operation under the guidance of digital subtraction angiography (DSA). Surgery was performed using a single lateral rectus abdominis approach combined with presacral decompression. The operation time, intraoperative blood loss and full weight-bearing time were recorded. The visual analogue scale (VAS) and European QOL Five Dimensional health scale (EQ-5D) were compared before and after operation. The Gibbons' impairment scale was used to assess neurological function. X-ray and CT scan were used to observe internal fixation and fracture healing. The complications during and after operation were recorded. Results:The patients were followed up for 24-52 months [(35.2±5.2)months]. The operation time was (2.9±0.6)hours. The intraoperative bleeding was (573±138)ml, and the full weight-bearing time was (11.6±1.2)weeks. X-ray and CT scan showed bone healing in all patients at the latest follow-up. The VAS and EQ-5D scale improved from preoperative (7.8±0.6)points and (0.34±0.07)points to the final follow-up of (0.8±0.3)points and (0.81±0.05)points ( P<0.05). According to Gibbons classification, 8 patients were grade I and 1 patient was grade II one year after operation ( P<0.01). Namely, the radiation pain of lower extremities was significantly improved in all patients, among which 8 patients showed pain disappeared and completely returned to normal and 1 patient showed residual numbness and hypoesthesia of the affected limbs. No major complications (eg, iatrogenic lumbosacral plexus injury, vital blood vessels or pelvic organs injury) occurred during the operation. During the follow-up period, only one patient developed traumatic hip arthritis and underwent total hip arthroplasty 6 months after operation. Fractures of the remaining patients were healed well without complications like infection, pressure ulcer or implant failure. Conclusions:For old Denis type II sacral fractures complicated with upper sacral plexus injury, lateral rectus abdominis approach combined with presacral decompression can fully decompress the upper sacral plexus nerve, relieve pain, and promote functional rehabilitation, with low incidence of complications. It is an alternative surgical method for the treatment of old Denis type II sacral fractures complicated with upper sacral plexus injury.

Bacitracin is a broad-spectrum cyclic peptide antibiotic, and mainly produced by Bacillus. Energy metabolism plays as a critical role in high-level production of target metabolites. In this study, Bacillus licheniformis DW2, an industrial strain for bacitracin production, was served as the original strain. First, our results confirmed that elimination of cytochrome bd oxidase branch via deleting gene cydB benefited bacitracin synthesis. Bacitracin titer and ATP content were increased by 10.97% and 22.96%, compared with those of original strain, respectively. Then, strengthening cytochrome aa3 oxidase branch via overexpressing gene qoxA was conducive to bacitracin production. Bacitracin titer and ATP content were increased by 18.97% and 34.00%, respectively. In addition, strengthening ADP synthesis supply is also proven as an effective strategy to promote intracellular ATP accumulation, overexpression of adenosine kinase DcK and adenylate kinase AdK could all improve bacitracin titers, among which, dck overexpression strain showed the better performance, and bacitracin titer was increased by 16.78%. Based on the above individual methods, a method of combining the deletion of gene cydB and overexpression of genes qoxA, dck were used to enhance ATP content of cells to 39.54 nmol/L, increased by 49.32% compared to original strain, and bacitracin titer produced by the final strain DW2-CQD (DW2ΔcydB::qoxA::dck) was 954.25 U/mL, increased by 21.66%. The bacitracin titer produced per cell was 2.11 U/CFU, increased by 11.05%. Collectively, this study demonstrates that improving ATP content was an efficient strategy to improve bacitracin production, and a promising strain B. licheniformis DW2-CQD was attained for industrial production of bacitracin.
Bacillus licheniformis ; metabolism ; Bacitracin ; biosynthesis ; Energy Metabolism ; genetics ; Industrial Microbiology ; methods