[Efficacy and safety of first-line treatment with anti-CD38 monoclonal antibody-based regimen for primary plasma cell leukemia].

Objective: To analyze the efficacy and safety of first-line treatment with an anti-CD38 monoclonal antibody regimen for primary plasma cell leukemia (pPCL). Methods: Patients diagnosed with pPCL from December 1st, 2018 to July 26th, 2023, receiving first-line treatment of anti-CD38 monoclonal antibody-based regimens across multiple centers including Peking University People's Hospital, Fuxing Hospital of Capital Medical University, Qingdao Municipal Hospital, Shengjing Hospital of China Medical University, Handan Central Hospital, the First Affiliated Hospital of Harbin Medical University, the Fourth Hospital of Hebei Medical University and General Hospital of Ningxia Medical University were consecutively included. A total of 24 pPCL patients were included with thirteen being male and eleven being female. The median age [M(Q1, Q3)] was 60 (57, 70) years. Patients were grouped according to peripheral blood plasma cell (PBPC) percentage [5%-19% (n=14) vs ≥20% (n=10)]. Last follow-up date was September 26th, 2023. The median follow-up period was 9.1 (4.2, 15.5) months. Patients' data related with clinical baseline characteristics, efficacy, survival and safety were retrospectively collected. Cox proportional hazards regression model was used to analyze risk factors associated with survival. Results: Among 24 pPCL patients, 16 (66.7%) patients had anemia at diagnosis, 13(54.2%) patients had thrombocytopenia, 8 (33.3%) patients had a baseline estimated glomerular filtration rate (eGFR)<40 ml·min-1·(1.73m2)-1, 13 (54.2%) patients had elevated lactate dehydrogenase (LDH) levels. The median PBPC percentage was 16% (8%, 26%) . Fluorescence in situ hybridization testing indicated that patients harboring 17p deletion, t(4;14) or t(14;16) were 6 (25.0%), 4 (16.7%) and 4 (16.7%), respectively. The overall response rate was 83.3% (20/24). The median progression-free survival (PFS) was 20.5 (95%CI: 15.8-25.2) months, and the median overall survival (OS) was not reached. Estimated 1-year and 2-year PFS and OS rates were 75.0% and 89.1%, 37.5% and 53.4%, respectively. The median PFS and OS for patients with PBPC percentages 5%-19% and≥20% were not reached and 20.5 (95%CI:15.7-25.3) months, 17.8 months and not reached, respectively. There was no significant statistical difference of PFS and OS between two groups (all P>0.05). Multivariate Cox regression analysis showed that 1p32 deletion was the risk factor associated with PFS (HR=7.7, 95%CI: 1.1-54.9, P=0.043). Seventeen patients (70.8%) developed grade 3-4 hematologic toxicities. Twelve patients (50.0%) developed grade 3-4 thrombocytopenia. Sixteen patients (66.7%) developed infection. All hematologic toxicities and infections were improved after supportive treatment. Conclusion: First-line treatment with anti-CD38 monoclonal antibody-based therapy for pPCL is effective and safe.

[Clinical effects of expanded frontal flap and flip scar flap in repairing partial nasal defect].

Objective: To investigate the clinical effects of expanded frontal flap and flip scar flap in repairing partial nasal defect. Methods: A retrospective observational study was conducted. From January 2012 to January 2022, 26 patients with partial nasal defects who met the inclusion criteria were admitted to the First Affiliated Hospital of Air Force Medical University, including 19 males and 7 females, aged 5 to 61 years. The surgery was performed in 4 stages. In the first stage, a rectangular skin and soft tissue expander (hereinafter referred to as expander) with suitable rated capacity was planted in frontal region and expanded by injecting water regularly. In the second stage, flip scar flap was grafted to reconstruct nasal inner lining, whose area was about 10% larger than the area of defect. The expanded frontal flap with pedicle was transferred to repair the nasal defect, whose pedicle was supraorbital vessel or supratrochlear vessel on the contralateral side of the defect, and the area of expanded flap was 20% larger than the nasal defect area after resection and flipping of scar flap. The donor site of expanded flap was sutured directly. After 3 weeks of flap transferring, the flap was delayed in the third stage. After 1 week of delaying operation, the pedicle of flap was cut off in the fourth stage. The number, rated capacity, injection volume, and expansion time of embedded expanders were recorded. The occurrences of complications including infection, hematoma, ulceration of expanded flap after the first stage operation, and blood supply disorder or necrosis of flap after operation in the second and fourth stages were observed. All the patients were followed up for 1 year at least, and the color of flap, scar of frontal donor site, symmetry of bilateral eyebrows, and the nasal appearance and ventilated function of external nasal tract were observed. Results: A total of 26 expanders were embedded in 26 patients. The rated capacity of expanders ranged from 100 to 300 mL. The injection volume was 1.0 to 1.5 times of the rated capacity of expanders. The expansion time ranged from 2.5 to 4.0 months, with an average time of 3 months. There were no complications occurred after each operation. The follow-up showed that the color of flap was similar to the normal nasal skin, the scar of frontal region was not obvious, the bilateral eyebrows were basically symmetrical, the nose had excellent appearance, ventilation function of external nasal tract was not affected, while some of the patients had downward rotation or unapparent tip-defining point of nose. Conclusions: Using the flip scar flap to reconstruct the nasal inner lining and pre-expanded frontal flap to reconstruct the nasal skin, without free cartilage transplantation to repair the partial nasal defects can achieve satisfied nasal appearance post operation, without abnormal external nasal ventilation function.

[An exploratory clinical study of the efficacy and safety of tumor-infiltrating lymphocytes in the treatment of metastatic osteosarcoma].

Objective: To explore the safety and efficacy of tumor-infiltrating lymphocytes (TILs) extracted from tumor tissue in patients with pulmonary metastasis of osteosarcoma, the TILs were amplified in vitro to reach clinical dosage and reinfused to the patients combined with high-dose interleukin 2 (IL-2). Methods: Twelve subjects with pathologically diagnosed osteosarcoma were enrolled from December 2019 to June 20, 2021 in Shanghai General Hospital. All subjects progressed with metastasis after standard chemotherapy and failed multiple lines of treatments. Fresh tumor tissue was obtained from the metastatic site and extracted and amplified by Good Manufacturing Practice (GMP) workshop to produce TILs to clinical treatment dosage (109-1011). High-dose IL-2 (100 000-200 000 U/kg) was administered immediately after autogenous TILs infusion to promote the activation, proliferation and antitumor cytolytic activity in vivo. Adverse events (AE) were graded according to Common Terminology Criteria for Adverse Events (CTCAE) standard and tumor response was assessed according to Response Evaluation Criteria in Solid Tumors (RECIST) 1.1. Results: One patient did not receive treatment due to failure in isolating TILs, total of 11 patients received a single re-infusion of autologous TILs. There were 10 males and 1 female with a median age of 19.9 years (12-33 years). Six of these patients received higher dose levels of 1.0×1010 TILs. The 11 patients were followed-up for 1 to 13 months and tolerated well. The most common adverse events reported were fever (10/11), constipation (3/11) and elevated gamma-glutamyl transferase (GGT) (3/11). The high incidence of fever was due to the IL-2 infusion. All patients experienced a transient drop in lymphocyte count and leukopenia leading to non-myeloid ablative lymphocyte clearance. The AE included grade 4 hematologic toxicity, including 8 cases of lymphocytopenia, 2 cases of neutropenia and 1 case of thrombocytopenia. No AE of neurotoxicity occurred. Of all the 11 patients, 9 patients got stable disease (SD) and 2 patients had progressive disease (PD). The disease control rate was 9/11. The median duration of SD was more than 4 months, and the maximum tumor volume decreased by close to 20%. Patient number 9 had sustained SD status for more than 6 months. Conclusions: TILs with in vitro expansion ability could be isolated from tumor tissues of advanced osteosarcoma patients. TILs amplified and reinfused in vitro have anti-osteosarcoma activity.

[Application of skin and soft tissue expansion in repairing pediatric patients with superficial defects].

Skin and soft tissue expansion can provide skin tissue similar to the recipient area in color and texture, which is one of the ideal methods in the repair of superficial defects. However, due to the long treatment cycle and relatively high complications rate in pediatric patients, expansion still faces many challenges. Based on the clinical practice and the current progress in skin and soft tissue expansion, this paper briefly discusses the change of skin after expansion, and the application, prevention and treatment of complications in the application of expansion in pediatric patients, aiming to provide reference for expansion in pediatric patients.

[Establishment and validation of a clinical prediction model for infection risk at the placement sites of skin and soft tissue expanders].

Objective: To establish a clinical prediction model for infection risk at the placement sites of skin and soft tissue expanders (hereinafter termed as expanders) and to validate the predictive value of the model. Methods: A retrospective observational study was conducted. Totally 2 934 patients who underwent skin and soft tissue dilatation surgery in the Department of Plastic Surgery of the First Affiliated Hospital of Air Force Medical University from January 2009 to December 2018 and met the selection criteria were included. There were 1 867 males and 1 067 females, with a median age of 18 years. Totally 3 053 skin and soft tissue expansion procedures were performed with 4 266 expanders implanted. The following indexes were selected as predictor variables, including patients' age, gender, marital status, ethnicity, hospital admission, surgical indication, disease duration, with/without history of smoking, history of drinking, history of blood transfusion, history of underlying diseases, and inability to use cephalosporin antibiotics due to allergy, number of expander in a single placement, rated volume of expander, water injection rate of expander in the first time, placement site of expander, anesthesia method, duration of operation, and with/without postoperative hematoma evacuation, and infection at the placement site of expander as the outcome variable. Univariate analysis of the data was performed using least absolute shrinkage and selection operator (LASSO) regression to screen the potential risk factors affecting infection at the placement sites of expanders, the factors selected by the univariate analysis were subjected to binary multivariate logistic regression analysis to screen the independent risk factors affecting infection at the placement sites of expanders, and a nomogram prediction model for the occurrence of infection at the placement sites of expanders was established. The C index and Hosmer-Lemeshow goodness of fit test were used to evaluate the discrimination and accuracy of the model, respectively, and the bootstrap resampling was used for internal verification. Results: The results of LASSO regression showed that age, gender, hospital admission, surgical indication, disease duration, history of drinking, history of heart disease, history of viral hepatitis, history of hypertension, inability to use cephalosporin antibiotics due to allergy, number of expander in a single placement, rated volume of expander, placement site of expander, postoperative hematoma evacuation were the potential risk factors for infection at the placement sites of expanders (regression coefficient=-0.005, 0.170, 0.999, 0.054, 0.510, -0.003, 0.395, -0.218, 0.029, 0.848, -0.116, 0.175, 0.085, 0.202). Binary multivariate logistic regression analysis showed that male, emergency admission, disease duration ≤1 year, inability to use cephalosporin antibiotics due to allergy, rated volumes of expanders ≥200 mL and <400 mL or ≥400 mL, and expanders placed in the trunk or the limbs were the independent risks factors for infection at the placement sites of expanders (odds ratio=1.37, 3.21, 2.00, 2.47, 1.70, 1.73, 1.67, 2.16, 95% confidence interval=1.04-1.82, 1.09-8.34, 1.38-2.86, 1.29-4.41, 1.07-2.73, 1.02-2.94, 1.09-2.58, 1.07-4.10, P<0.05 or P<0.01). The C index for evaluating the discriminative degree of the model was 0.63, the Hosmer-Lemeshow goodness of fit test for evaluating the accuracy of the model showed P=0.685, and the C index for internal validation by the bootstrap resampling was 0.60. Conclusions: Male, emergency admission, disease duration ≤1 year, inability to use cephalosporin antibiotics due to allergy, rated volume of expander ≥200 mL, and expanders placed in the trunk or the limbs are the independent risk factors for infection at the placement sites of expanders. The clinical prediction model for infection risk at the placement sites of expanders was successfully established based on these factors and showed a certain predictive effect.

[Clinical effects of single pedicle transfer of expanded axial flap across the midline of the frontal-parietal region in reconstruction of large scar deformities in the face and neck].

Objective: To explore the clinical effects of single pedicle transfer of expanded axial flap across the midline of the frontal-parietal region in reconstruction of large scar deformities in the face and neck. Methods: From January 2016 to August 2019, 10 male patients, aged from 20 to 52 years with post-burn facial and cervical scar deformities, were admitted to the First People's Hospital of Zhengzhou, with the size of scar ranging from 15 cm×7 cm to 23 cm×11 cm. In the first stage, a cylindrical skin and soft tissue expander with rated capacity ranging from 400 to 600 mL was placed in the frontal-parietal region. Another cylindrical expander with rated capacity ranging from 50 to 100 mL was placed in the temporal region of the patient with scars in front of the ear and in cheek. The injection time was 3 to 5 months with the total injection volume being 1.5 to 2.5 times of the rated capacity of expander. In the second stage, the superficial temporal artery frontal branch and its branches were explored, the expander was removed, the scars in the face and neck were conducted resection and contracture relaxation, and the single pedicle transfer of expanded axial flap across the midline of the frontal-parietal region for reconstruction was performed. When the branches of the superficial temporal vessels were difficult to be detected by Doppler ultrasonic blood stream detector, the patient underwent computed tomography (CT) angiography and three-dimensional reconstruction. The donor site in frontal-parietal region was directly sutured, and the wound of the exposed donor site at the pedicle and temporal region was temporarily covered with scar skin. After the suture wound was healed and the hair in expanded flap grew out, hair removal and laser hair removal were performed. Three to four weeks after transplantation of expanded flap, the flap pedicle was cut off, restored, and trimmed in the third stage. The status about the completion of operation, the implantation of expander in the temporal region, CT angiography and three-dimensional reconstruction were recorded. The effective resection area of expanded flap, the length across the midline and the length of the pedicle, and the survival status of the expanded flap and complications after operation were observed. The appearance of donor and recipient sites, the scar recurrence, the appearance and function improvement of patients, and the satisfaction degree of patients were followed up. Results: All the 10 patients successfully completed three stages of operation, of which 6 patients had an auxiliary expander placement in the temporal region, and 5 patients underwent CT angiography and three-dimensional reconstruction. The effective resection area of expanded flap ranged from 18 cm×8 cm to 25 cm×13 cm. The distal end of the flap across the midline extended 4-6 cm to the opposite side, and the length of pedicle was 2-6 cm. All the expanded flaps of patients survived well after formation and transfer. The venous reflux disorder and obvious swelling occurred in 6 patients at the distal end of the flap after operation, and the blood supply recovered after acupuncture bloodletting, etc. Follow-up of 6 to 24 months showed that the color, texture, and thickness of the expanded flaps were similar to those of the facial skin, and no recurrence of scar was observed; the incision in the donor site of the frontal-parietal region was concealed, the hair growth of the temples and head was normal, and the reconstructed hairline was natural; compared with those before operation, the appearance, head-up, mouth-opening, and other functions of patients were significantly improved; the patients were satisfied with the effect of reconstruction. Conclusions: Clinical application of single pedicle transfer of expanded axial flap across the midline of the frontal-parietal region in reconstruction of large scar deformities in the face and neck can achieve a good appearance and function, and the donor site shows good shape, which enriches the application range of the trans-regional blood supply flap. It is a reliable method for reconstruction of large scar deformities in the face and neck.

[Isolated biceps tenodesis by double row for pulley lesions].

Objective: To explore clinical outcome of isolated arthroscopic biceps tenodesis by double row for pulley lesions. Methods: Forty-nine patients with pulley lesions were treated from July 2017 to June 2018 in the Department of Sport Medicine, the Affiliated Zhongshan Hospital of Dalian University by isolated arthroscopic biceps tenodesis by double row. Patients were divided into 2 groups according to the intraoperative damage of the pulley system. In group A, there were 16 patients with isolated superior glenohumeral ligament/coracohumeral ligament (SGHL/CHL) complex lesions, including 9 males and 7 females, aged (55±6) years. In group B, there were 33 patients (15 males and 18 females, aged (57±8) years) with SGHL/CHL complex and adjacent supraspinatus tendon and/or subscapularis tendon articular-side partly tears. Patients in two groups were treated with different isolated arthroscopic biceps tenodesis by double row. Constant-Murley shoulder score and pain visual analogue scale (VAS) score were assessed before operation and 3, 6, 12 months after the operation. Postoperative complications were also recorded in two groups. The t test was used to compare the quantitative data within and between two groups. Results: All 49 patients were followed up for 12 to 24 months with an average of (17±6) months. The first-stage healing was achieved in all incisions in the two groups. No surgical complications related to revision, infection, Popeye syndrome and cramping pain were observed in either group. There was 1 case treated by secondary arthroscopy for retrauma in group B. The Constant-Murley shoulder score in group A before the operation was 46±10, and it was increased to 89±9 at the 12 months post operation(t=-22.637, P<0.05); and it was 39±10 and 87±8 before and 12 months after the operation respectively in group B (t=-44.849, P<0.05). The VAS scores in the two groups were both decreased significantly at the 12 months post operation when compared with those before the operation (0.68±0.70 vs 5.25±0.27 and 0.72±0.83 vs 5.69±0.84, respectively) (t=29.007, 37.079, both P<0.05). Conclusion: Isolated arthroscopic biceps tenodesis by double row can relieve pain, recover functions of shoulder joint effectively, and achieve a satisfactory outcome in the treatment of pulley lesions.

[Clinical effect of pre-expanded deltopectoral flap in the repair of faciocervical lesion and defect].

Objective: To explore the clinical effect of pre-expanded deltopectoral flap in the repair of faciocervical lesion and defect. Methods: From July 2004 to August 2018, 355 patients with faciocervical lesion and defect were admitted to the First Affiliated Hospital of Air Force Medical University, including 200 males and 155 females aged 4 to 48 years with major conditions including thermal burn scars, and type Ⅲ and Ⅳ facial-cervical deformities. During the stage Ⅰ skin soft tissue expander implantation surgery, according to the size and location of lesion and defect, expanders with appropriate volume were placed to expand the deltopectoral area. During the stage Ⅱ flap pedicled transposition surgery, after the expander was expanded to the desired volume, the impairment tissue was removed, the flap was designed according to the size of the defect (the unilateral defect area was 7 cm×5 cm to 17 cm×16 cm) and pedicled transposition was carried out. The incision in the chest donor area was directly sutured and closed. After the flap survived, stage Ⅲ flap delay and pedicle division surgery was carried out. The area of one single flap was 8 cm×5 cm to 20 cm×18 cm. The numbers of flaps and expanders, rated volume and expansion of expander, the intervals between surgeries in each stage, flap survival, postoperative complications in surgeries in each stage, and follow-up were recorded and analyzed. Results: A total of 460 pre-expanded deltopectoral flaps were used, including 250 unilateral flaps and 105 bilateral flaps. Totally 460 expanders were used in this group of patients. The rated volume was mostly 500 mL (163 expanders) and 600 mL (142 expanders). The expansion multiple of the expander was (1.14±0.19) times of the rated volume. The flap expansion time of the patients was (96±30) d, the pedicle time was (32±8) d, and the delay time was (7.5±1.6) d. The postoperative complications of patients mainly included infection (29 patients), expander exposure (18 patients), and hematoma (10 patients). During the follow-up of 6 to 120 months, the elasticity, texture, and color of the flaps of patients were similar to the surrounding tissue of the recipient area, and the face and neck were symmetrical, not bloated. Conclusions: The deltopectoral flap obtained by overexpansion has a larger area and a thinner thickness, and the elasticity, texture, and color are similar to the surrounding tissue of the recipient area. After transfer, a stable appearance of the face and neck can be obtained. The main complications are infection and expander exposure, most of which occurred after stage Ⅰ skin soft tissue expander implantation surgery.

[Clinical effects of expanded forehead flaps in repairing midfacial defects].

Objective: To explore the clinical effects of expanded forehead flaps in repairing midfacial defects. Methods: From January 2003 to December 2018, 19 patients with midfacial defects were admitted to our unit, including 8 males and 11 females, aged 7 to 52 years. One cylindrical expander with rated capacity ranged from 100 to 170 mL was placed in the forehead of patients in the first stage of expansion, and the total water injection volume was about 2 times of the rated capacity of the expander during 1 to 2 months. The area of midfacial defects was 4 cm×2 cm to 9 cm×5 cm after resection in the second stage surgery. Expanded forehead flaps with vascular pedicle of supratrochlear vessels or frontal branch of superficial temporal vessels were used to repair the midfacial defects, with flap size ranging from 5 cm×2 cm to 16 cm×6 cm. The donor sites were closed by direct suturing. Three weeks later, the pedicle was divided. The complications, blood supply after flap transfer and pedicle division, and the treatment effects during follow-up were observed. Results: Among the patients, flaps of 11 patients had vascular pedicle of supratrochlear vessels; flaps of 8 patients had vascular pedicle of frontal branch of superficial temporal vessels. All the flaps survived with no complications and good blood supply after flap transfer and pedicle division. During the follow-up of 6 to 12 months after the third stage surgery of pedicle division of 12 patients, no lower eyelid ectropion occurred, the appearance of the flaps was similar to the surrounding tissue with no swelling. Conclusions: The application of expanded forehead flaps can not only repair the defects but also effectively avoid the complication of lower eyelid ectropion, which is a promising method in repairing midfacial defects.

[Role of PI3K/Akt signaling pathway in ischemic rats underwent cardiac shock waves therapy].

Objective: To investigate the role of PI3K/Akt signaling pathway in ischemic rats underwent cardiac shock therapy. Methods: Adult male Sprague Dawley (SD) rats weighing 220-250 g were used to establish a heart failure model by ligation of the left anterior descending coronary artery. Rat models were defined by echocardiographic assessment at 4 weeks post operation and heart failure rats were randomly divided into 4 groups,namely heart failure group (HF group, 9 cases),heart failure+cardiac shock waves therapy group (HF+CSWT group, 9 cases),heart failure+inhibitor(HF+LY294002 group, 9 cases),heart failure+cardiac shock waves therapy group+inhibitor (HF+CSWT+LY294002 group, 9 cases),and another 9 sham-operated SD rats served as control group (sham group, 9 cases). At 8 weeks postoperation, echocardiography was used to evaluate cardiac function in each group,myocardial infarct size was measured by TTC staining,the apoptotic index of rats cardiomyocytes were detected by TUNEL method,the myocardial mRNA expression of apoptosis-related factor was detected by real-time quantitative PCR, the protein expression levels of PI3K/Akt signaling pathway and apoptosis-related pathways were detected by Western blot. Results: (1) Eight weeks after operation, left ventricular end diastolic diameter (LVEDD) and left ventricular end systolic diameter (LVESD) were significantly lower in HF+CSWT group than in HF group (all P<0.05), left ventricular ejection fraction (LVEF) and left ventricular shortening rate (LVFS) were significantly higher in HF+CSWT group than in HF group (all P<0.05),LVEF was significantly lower in the HF+ CSWT+ LY294002 group than in HF+ CSWT group (P<0.05). (2) Myocardial infarct size was significantly lower in the HF+ CSWT group than in HF group ((5.57 ± 0.51)% vs. (25.56 ± 0.56)%, P<0.05), which was significantly higher in the HF+CSWT+LY294002 group than in HF+CSWT group ((12.90±2.34)% vs. (5.57±0.51)%,P<0.05). (3) The cardiomyocyte apoptotic index was significantly lower in the HF+CSWT group than in the HF group ((30.25±6.12)% vs. (53.85±9.89)%,P<0.05), which was significantly higher in the HF+CSWT+LY294002 group than in the HF+CSWT group ((46.12±3.42)% vs.(30.25±6.12)%,P<0.05). (4) The myocardial mRNA expression of Bcl-2 was significantly higher, while myocardial mRNA Bax and Caspase-3 expression were significantly lower in HF+CSWT group than in HF group and HF+CSWT+LY294002 group (all P<0.05). (5) The expression levels of p-Akt, Bcl-2 and pro-Caspase-3 in myocardial tissue were significantly higher in the HF+CSWT group than in the HF group and HF+CSWT+LY294002 group (all P<0.05), which were significantly lower in the HF+LY294002 group than in the HF and HF+CSWT+LY294002 groups (all P<0.05). Myocardial Bax protein expression was significantly lower in the HF+CSWT group than in the HF group and the HF+CSWT+LY294002 group (all P<0.05), which was significantly higher in the HF+LY294002 group than in the HF group (P<0.05). Conclusion: CSWT improves cardiac function and inhibits cardiomyocyte apoptosis through PI3K/Akt signaling pathways in this rat HF model.

Influence of roflumilast on sepsis mice through the JAK/STAT signaling pathway.

OBJECTIVE: The aim of this study was to explore the influence of roflumilast on sepsis mice through the Janus kinase/signal transducer and activator of transcription (JAK/STAT) signaling pathway. MATERIALS AND METHODS: A total of 36 Sprague-Dawley mice were randomly divided into normal group (n=12), model group (n=12) and roflumilast group (n=12). Mice in the normal group were fed normally. However, mice in the model group and roflumilast group were intraperitoneally injected with endotoxin to establish the sepsis mouse model. Furthermore, mice in the model group and roflumilast group were intraperitoneally injected with 0.9% sodium chloride and roflumilast once a day, respectively. After 7 d of intervention, mice were sampled. Lung tissue morphology was observed via hematoxylin-eosin (HE) staining, and the pathological score was given. The protein expression levels of JAK and STAT-3 were detected via Western blotting. The expression levels of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) were detected via enzyme-linked immunosorbent assay (ELISA). Meanwhile, the mRNA expression levels of JAK, STAT-3, IL-6 and TNF-α were detected via quantitative Polymerase Chain Reaction (qPCR). The number of inflammatory cells in the lavage fluid was counted by a biochemical detector. RESULTS: The survival rate of mice in the roflumilast group was significantly higher than that of the model group (p<0.05). The results of HE staining revealed that lung tissue morphology in roflumilast group was significantly improved when compared with the model group. Meanwhile, the pathological score in the roflumilast group was significantly lower than that of the model group (p<0.05). Western blotting showed that the protein expression levels of JAK and STAT-3 in the roflumilast group were markedly lower than those of the model group (p<0.05). According to the results of ELISA, the expression levels of IL-6 and TNF-α in the roflumilast group were remarkably lower than the model group (p<0.05). Further qPCR results manifested that the mRNA expression levels of JAK, STAT-3, IL-6 and TNF-α in the roflumilast group were significantly lower than those of the model group (p<0.05). Moreover, the number of neutrophils, monocytes and lymphocytes in the roflumilast group was significantly smaller than the model group. CONCLUSIONS: Roflumilast can improve lung tissue morphology of sepsis mice by inhibiting the JAK/STAT signaling pathway.

UPK1B promotes the invasion and metastasis of bladder cancer via regulating the Wnt/ß-catenin pathway.

OBJECTIVE: The aim of this study was to investigate the expression of UPK1B in bladder cancer (BCa), and to further explore the correlation between UPK1B expression and pathological parameters as well as the prognosis of BCa. PATIENTS AND METHODS: Quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the expression of UPK1B in 92 pairs of BCa tissues and adjacent normal tissues. The relationship between UPK1B expression and pathological features as well as the prognosis of BCa patients was further analyzed. For in vitro experiments, the mRNA expression level of UPK1B in BCa cell lines (EJ and T-24) was detected by qRT-PCR. In addition, knockdown of UPK1B in BCa cells was constructed using small interfering RNA. Effects of UPK1B knockdown on biological functions of BCa cells were analyzed by Cell Counting Kit-8 (CCK-8), colony formation assay and transwell assay, respectively. Furthermore, the underlying mechanism of UPK1B in regulating BCa was evaluated by Western blot and qRT-PCR, respectively. RESULTS: The expression of UPK1B in BCa tissues was remarkably higher than that of adjacent normal tissues (p<0.05). Compared with BCa patients with lower UPK1B expression, those with higher UPK1B expression exhibited higher tumor stage, lymph node metastasis and distant metastasis. In vitro experiments indicated that cell proliferation, invasion and metastasis were remarkably decreased in cells transfected with si-UPK1B when compared with those transfected with negative controls. Western blot showed that the expression of key proteins in the Wnt/ß-catenin signaling pathway in cells transfected with si-UPK1B was significantly down-regulated compared with those transfected with negative controls, including ß-catenin, c-myc and cyclinD1. In addition, rescue experiments found that UPK1B was regulated by ß-catenin. CONCLUSIONS: UPK1B is upregulated in BCa, and is significantly correlated with tumor stage, lymph node metastasis, distant metastasis and poor prognosis of BCa. Moreover, UPK1B promotes the proliferation, invasion and migration of BCa via regulating the Wnt/ß-catenin signaling pathway.

[The impact of male circumcision on the natural history of genital HPV infection: a prospective cohort study].

Objective: To analyze the correlation between circumcision and incidence and clearance of male genital HPV infection. Methods: From May to July 2014, 18-55 year old men who had sexual behavior history were recruited from the general population in Liuzhou, Guangxi to set up a cohort. Totally, 113 circumcised and 560 uncircumcised men were enrolled and interviewed using a questionnaire (including information on demographic characteristics and sexual behaviors), then they were followed-up with 6-month interval for 2 times. On each visit, specimens of male external genitalia were collected and genotyped for HPV DNA. The differences of incidence and clearance of genital HPV infections between circumcised and uncircumcised men were analyzed by Log-rank test. Cox regression was used to analyze the relationship between circumcision and incidence and clearance of HPV infection. Results: The median age (P(25), P(75)) of circumcised and uncircumcised men were 28 (24, 35) and 32 (24, 31), respectively. The incidences of any HPV infections were 9.1 (95%CI: 2.4-15.7) and 8.4 (95% CI: 5.6-11.2) per 1 000 person-months (χ(2)=0.10, P=0.758), respectively. The clearance of circumcised men [136.3 (95%CI: 70.0-202.7) per 1 000 person-months] was higher than that in uncircumcised men [89.6 (95%CI: 65.9-113.3) per 1 000 person-months] (χ(2)=8.19, P=0.004). In multivariate COX regression analysis, compared with uncircumcised men, circumcised men had higher possibility to clear any HPV infections (HR: 2.41, 95%CI: 1.30-4.46). Compared with men having one sexual partner, people having more than 4 sexual partners had lower possibility to clear any HPV infections (HR: 0.49, 95%CI: 0.25-0.96). Compared with 18-25 years old men, men aged 26-35 years old had higher possibility to clear high-risk HPV infections (HR: 2.14, 95%CI: 1.08-4.23). Conclusion: Circumcised and uncircumcised men had similar incidence of genital HPV infection, whereas, men conducted circumcision and having fewer sexual partners could increase the clearance of genital HPV infections.

[The expressions of VEGF and VEGFR signaling pathway in the bone marrow mononuclear cells with chronic mountain sickness].

Objective: This study was aimed at investigating the levels and relationships of vascular endothelial growth factor (VEGF) and its receptor(VEGFR) in the bone marrow mononuclear cells (MNC) of chronic mountain sickness (CMS). Methods: A total of 34 patients with CMS and 30 controls residing at altitudes of 3 000-4 500 m were recruited for this study. The levels of VEGF, VEGFR1 and VEGFR2 in bone marrow MNC were detected by flow cytometry technique and RT-qPCR. Results: The percentage of VEGFR2 positive cells in the bone marrow MNC of CMS were higher than that of the controls[20.7% (8.1%, 67.6%) vs 8.1% (2.2%, 14.9%), P<0.05], but that of VEGFR1-positive and VEGF-positive were similar in CMS and controls. The mRNA levels of VEGFR2 were higher in the bone marrow MNC of CMS than in the controls[1.7(1.0, 5.1) vs 1.0(0.4, 2.7), P<0.05], while VEGF and VEGFR1 mRNA levels were similar between the two groups. The percentage of VEGFR2 positive cells in CMS were significantly correlated with hemoglobin (r=0.453, P=0.007) and the percentage of VEGF-positive cells (r=0.373, P=0.030). Conclusions: Bone marrow MNC of CMS may show enhanced activity of the VEGF-VEGFR2 pathway, and it appears to be involved in the pathogenesis of CMS.

[Sivelestat alleviates nonalcoholic steatohepatitis in mice through inhibiting activation of Kupffer cells].

Objective: To investigate the role of neutrophil elastase inhibitor, sivelestat, in preventing and treating nonalcoholic steatohepatitis (NASH) and its underling mechanisms. Methods: A total of forty 4-week-old male C57BL/6J ApoE-/-mice were equally divided into the following four groups: standard chow (SC)+isotonic saline; SC+sivelestat; high-fat, high-cholesterol (HFHC) diet+isotonic saline; and HFHC+sivelestat. These mice were treated with above methods for 12 weeks. Blood and liver tissue samples were collected to measure biochemical parameters, hepatic steatosis and non-alcoholic fatty liver disease (NAFLD) activity score (inflammation) were evaluated by oil red O staining and HE staining, respectively. The mRNA and protein expression levels of hepatic inflammatory cytokines, CD68, and F4/80 were determined by quantitative RT-PCR and immunohistochemistry, respectively. Comparison of means between the four groups was made by one-way analysis of variance, and comparison between any two groups was made by the LSD or SNK method (for data with homogeneity of variance) or the Tamhane or Dunnett method (for data with heterogeneity of variance). Results: Mice fed with an HFHC diet for 12 weeks developed typical pathological features of NASH compared with those fed with SC. Compared with mice fed with HFHC diet without sivelestat, those treated with HFHC and sivelestat exhibited the following features: (1) significantly reduced fast blood glucose, blood cholesterol, and hepatic biochemical parameters, as well as increased insulin sensitivity; (2) significantly reduced NAFLD activity score (5.71±1.11 vs 3.16±1.16, P < 0.05); (3) reduced monocyte chemoattractant protein-1 and tumor necrosis factor -α; (4) significantly reduced mRNA levels of CD68 and F4/80; and (5) reduced expression of CD68 in the liver. Conclusion: Sivelestat alleviates the hepatic steatosis and inflammation of NASH in mice by inhibiting the activation of Kupffer cells.

[The limitation of transthoracic echocardiography in diagnosing partial anomalous pulmonary venous drainage].

Objective: To improve the diagnostic accuracy of transthoracic echocardiography (TTE) by analyzing its limitations in diagnosing partial anomalous pulmonary venous drainage (PAPVD). Method: This was a retrospective analysis of PAPVD patients seen at the Children's Hospital of Fudan University from October 1 2006 to October 1 2016. The echocardiographic data were compared to findings on multi-slice spiral CT (MSCT), cardiac catheterization or surgery. The echocardiography machines used were Philip IE33, GE Vivid 7 and Vivid i with frequency ranging from 5.0 MHz to 7.5 MHz. The cardiac structure was analyzed according to Van Praagh segments. Result: A total of 43 cases of PAPVD were enrolled, male∶ female ratio 20∶23 with average age (27.9±21.4) months. Among them, 3 cases were simple PAPVD and 40 cases had other associated congenital heart diseases. TTE was successful in diagnosing 29 cases (67%) while 14 cases were missed. The diagnostic rate for right pulmonary vein drainage into superior vena cava, right atrium, inferior vena cava were 5/10, 17/20, and 3/5 respectively while left pulmonary vein drainage into left innominate vein was only 1/4. Added TTE images to re-exam the 9 of the 14 missed cases, 5 cases of abnormal drainage from right superior pulmonary vein were diagnosed, while 4 cases of drainage from right lower or left pulmonary vein were only picked up by indirect signs. Conclusion: The distance of the pulmonary veins from the routine ultrasound view and the possibility of branch number variation may limit the accuracy of TTE in diagnosing PAPVD, especially for drainage from right lower and left pulmonary vein. But TTE is still the preferred diagnostic method. The diagnostic rate could be increased by paying special attention to non-routine views including the suprasternal fossa, the right parasternal and subcostal area.

[Assessment of undiagnosed critical congenital heart disease before discharge from the maternity hospital].

Objective: Undiagnosed critical congenital heart disease (CCHD) was assessed before discharge from maternity hospital.Basic information was provided for screening CCHD in the early neonatal stage.Chi-squared test was used for comparison of categorical variables(detection rate of different types of CCHD). Method: A retrospective cohort study was conducted in neonates with CCHD who were admitted to Children's Hospital of Fudan University between 1 January 2012 and 31 December 2015. For comparing with the previously reported undiagnosed rate of CCHD at discharge, CCHD was defined as all duct dependent congenital heart disease (DDCHD) and any cyanotic CHD that required early surgery. Result: A total of 1 036 infants with CCHD were included. The prenatal detection rate of CCHD was 14.04%(122/869). As a whole, 52.51% (544/1 036) of CCHD cases were undiagnosed at discharge, and 14.09%(146/1 036)were still missed after 6-week examination. The diagnoses most likely to be unrecognized at discharge included critical coarctation of the aorta (COA) (75.00%), total anomalous pulmonary venous connection (61.54%), pulmonary atresia (PA) with ventricle septal defect (VSD) (61.45%), single ventricle (SV) (60.10%) and critical aortic stenosis (52.94%). Among newborns diagnosed prior to discharge, 54.88% (270/492) due to symptom or prenatal ultrasonographic diagnosis, 45.12% (222/492) due to abnormal findings in routine examination. Among asymptomatic CCHD cases without prenatal diagnosis, 71.02% (544/766) were undiagnosed and the most common delayed diagnosis was SV (82.78%), interrupted aortic arch (81.82%), transposition of the great arteries with intact ventricular septum (79.63%), PA/VSD (79.07%), and critical COA (78.57%). Newborns with DDC were more likely to develop symptoms within the first few days after birth, in comparison with non-DDC cases. However, their detection rates were close to each other. Conclusion: The rate of misdiagnosis of CCHD before discharge from maternity hospitals is high in China, indicates the importance of implementation of CCHD screening in Chinese maternity hospitals, so as to give timely diagnosis and proper treatment.

[Aesthetic reconstruction strategy for postburn facial scar and its clinical effect].

OBJECTIVE: To explore the aesthetic reconstruction strategy for postburn facial scar and its clinical effect. METHODS: Three hundred and forty-two patients with postburn facial scars were hospitalized from January 2000 to December 2015. Local expanded flap or deltopectoral expanded flap was used for reconstruction according to the location and size of the facial scar. The forehead expanded flap could be chosen for the scar in dorsum nasi or inferior eyelid. The local expanded flap was chosen when the scar width was smaller than 5 cm in cheek, chin, and marginal mandible region. The expanded deltopectoral flap was chosen when the scar width was larger than 5 cm in cheek, chin, and marginal mandible region or the scar contracture was too serious to cause displacement of lips, nose, or eyelid, and the wound width was larger than 5 cm after release. The facial scars of 82 patients, with size ranged from 6.0 cm×2.5 cm to 15.0 cm×10.0 cm, were reconstructed with expanded local flaps. The facial scars of 260 patients, with size ranged from 8.0 cm×7.0 cm to 38.0 cm×13.0 cm, were reconstructed with expanded deltopectoral flaps. After expansion of 2 to 6 months, the facial scars were excised and completely released first of all. The transfer way of local flap and size of deltopectoral flap with pedicle were designed according to the size and shape of the wound. Three weeks after transfer of deltopectoral flap, flap delay procedure was conducted. One week later, the pedicle was severed from the flap to reconstruct the remaining scar. Anti-scar medicine, laser therapy, and elasticized fabric were used postoperatively on the scars in both donor and recipient sites. RESULTS: During the postoperative follow-up for 3 to 12 months, the flaps of 40 out of 82 cases reconstructed with expanded local flaps were in good color and texture. Before 2008, mild scar hyperplasia was observed in the incision of 19 patients; with application of laser after 2008, the number of patients with scar hyperplasia was decreased. During the postoperative follow-up for 3 to 12 months, the flaps of 90 out of 260 cases reconstructed with expanded deltopectoral flaps were in good color and texture. The expander was exposed from the incision in 15 patients, while it did not affect the later treatment. Nine unilateral flaps showed poor blood circulation at the distal end, and they were healed after dressing change. In the early phase, necrosis was observed in one flap after transfer, and it was healed after transplantation of free skin graft. Scar hyperplasia was observed in the chest donor site of one patient, and it was improved after laser therapy. CONCLUSIONS: Postburn facial scar could be reconstructed with local or deltopectoral flaps, following the principle of similarity. The expansion could increase the size of the flaps, reduce the thickness of the flaps, and lower the donor site damage.

[Identification and verification of the candidate proteins that interact and collaborate with ATF3 in inhibiting hepatocarcinogenesis].

OBJECTIVE: To identify and verify proteins that interact and collaborate with ATF3 in inhibiting hepatocarcinogenesis. METHODS: Immunoprecipitation (IP), co-IP and protein spectrum analysis were used to identify the protein which interacted with ATF3 in HepG2. Immunohistochemistry (IHC) and Western blot (WB) were used to detect the expression pattern of ATF3 and its candidate interacting proteins in liver tissue. RESULTS: The protein expression differences were detected by IP in two HepG2 groups. The experimental group was infected by lentiviral vector with ATF3 over-expression and the control group was infected by mock-vehicle. Several protein bands with expression diversity were analyzed by protein spectrum, which revealed several candidate proteins that may be related with ATF3. Peptide sequences were analyzed by Mascot software and NCBI database. Combined with the existing literature and our study results, Gelsolin (GSN) was identified as a protein closely interacting with ATF3 and confirmed by co-IP, IHC and WB. CONCLUSIONS: GSN is identified and verified as an interacting protein with ATF3. ATF3 may function as a suppressor of liver cancer via protein-protein interactions with Gelsolin.