OBJECTIVE To establish characteristic chromatogram of Yao medicine Kadsura longipedunculata and the method for the content determination of its main component anwulignan， and evaluate the anti-inflammatory activity of anwulignan. METHODS HPLC method was performed with acetonitrile-0.5% phosphoric acid solution as the mobile phase for gradient elution. The characteristic chromatogram of K. longipedunculata was established and similarity was evaluated by Similarity Evaluation System for Chromatographic Fingerprint of TCM （2012 edition）. The content of anwulignan in K. longipedunculata was determined. Lipopolysaccharide induced RAW264.7 macrophages were selected as inflammatory cell model to investigate the effects of anwulignan on the levels of tumor necrosis factor-α （TNF-α）， interleukin-1β （IL-1β） and IL-6. RESULTS The similarities of characteristic chromatogram for 10 batches of K. longipedunculata ranged 0.901-0.994， and 9 common peaks were determined； 3 components were identified， such as changnan schisantherin E， kadsulactone A， anwulignan. The contents of anwulignan were （0.72±0.05）-（1.21±0.03） mg/g（n＝3）. Anwulignan of 0.125-0.5 μg/mL greatly decreased the levels of TNF-α， IL-1β and IL-6 in the supernatant of inflammatory model cells （P＜0.05 or P＜0.01）. CONCLUSIONS HPLC characteristic chromatogram of K. longipedunculata and the method for the content determination of anwulignan are all established， and anwulignan may be the active ingredient of anti-inflammatory effect in K. longipedunculata.

Infectious bone defect is bone defect with infection or as a result of treatment of bone infection. It requires surgical intervention, and the treatment processes are complex and long, which include bone infection control,bone defect repair and even complex soft tissue reconstructions in some cases. Failure to achieve the goals in any step may lead to the failure of the overall treatment. Therefore, infectious bone defect has been a worldwide challenge in the field of orthopedics. Conventionally, sequestrectomy, bone grafting, bone transport, and systemic/local antibiotic treatment are standard therapies. Radical debridement remains one of the cornerstones for the management of bone infection. However, the scale of debridement and the timing and method of bone defect reconstruction remain controversial. With the clinical application of induced membrane technique, effective infection control and rapid bone reconstruction have been achieved in the management of infectious bone defect. The induced membrane technique has attracted more interests and attention, but the lack of understanding the basic principles of infection control and technical details may hamper the clinical outcomes of induced membrane technique and complications can possibly occur. Therefore, the Chinese Orthopedic Association organized domestic orthopedic experts to formulate An evidence-based clinical guideline for the treatment of infectious bone defect with induced membrane technique ( version 2023) according to the evidence-based method and put forward recommendations on infectious bone defect from the aspects of precise diagnosis, preoperative evaluation, operation procedure, postoperative management and rehabilitation, so as to provide useful references for the treatment of infectious bone defect with induced membrane technique.

Objective:To observe and analyze the multimodal imaging characteristics of fundus in patients with sympathetic ophthalmia (SO).Methods:A retrospective study. From October 2012 to December 2021, 28 patients (36 eyes) diagnosed SO in the Department of Ophthalmology, Beijing Tongren Hospital were inclued in the study. There were 19 males (25 eyes) and 9 females (11 eyes), with the mean age of 51.61±12.02 years. There were 8 exciting eyes and 28 sympathizing eyes. The time to onset after trauma or surgery was 46.10±107.98 months. All patients underwent examinations including vision test, color fundus photograph, optical coherence tomography (OCT), fundus fluorescence angiography (FFA), indocyanine green angiography (ICGA). Angio-OCT (OCTA) was performed on 3 eyes and fundus autofluorescence (AF) was performed on 8 eyes. The early and late phase were defined respectively as ≤2 months and >2 months. Their multimodal imaging characteristics were summarized.Results:In 8 exciting eyes, subretinal fibrosis with mutifocal retinal atrophy and pigmentation was noted in 5 eyes (62.50%, 5/8), the other 3 eyes showed sunset glow fundus (37.50%, 3/8). In 28 sympathizing eyes, in the early phase, the fundus photograph showed shallow retinal detachment with optic disc edema in 9 eyes (32.14%, 9/28); in the late phase, peripapillary yellowish-white subretinal lesions in 11 eyes (39.29%, 11/28). In the late course of the disease, there were yellow-white lesions around the optic disc (peridisc) and peripheral subretinal area in 11 eyes (39.29%, 11/28). Dalen-Fuchs nodules were found in 10 eyes (35.71%, 10/28). On OCT, multiple serous retinal detachment and irregular choroidal folds were noted in the early phase; hill-like subretinal hyperreflective elevation was noted in peripapillary area and subfovea with presence of cystic spaces in the intraretina in the late phase. FFA examination showed"pinpoint-like" strong fluorescence in the early stage, and "multi-lake-like" fluorescein accumulation and leakage in the late stage; "map-like" weak fluorescence around the disc in the early stage of the disease, dot-like strong fluorescence lesions in each quadrant of the peripheral retina, and fluorescence in the late stage of the disease course. enhanced. ICGA examination showed that the FFA strong fluorescence lesions in the middle and late stages were weak fluorescence. FAF examination, point-like strong and weak autofluorescence lesions with unclear boundaries. Nine sympathizing eyes with subretinal yellow-white lesions has vision without light-0.1 (significantly decreased vison), while 8 eyes with sunset glow fundus was 0.5-1.0 (mildly decreased vison).Conclusions:SO could not only show the semblable features of acute phases of Vogt-Koyanagi-Harada syndrome, but also the yellowish-white lesions in the peripapillary area, macula and periphery. Most of the eyes with peripapillary lesions has a significantly decreased vison, while the eyes with sunset glow fundus has a mildly decreased vison.

Objective: To investigate the effects of c-Met inhibitor AMG-102 on the proliferation and apoptosis of laryngeal squamous carcinoma Hep-2 cells and the underlying mechanism. Methods: Laryngeal squamous carcinoma cell line Hep-2 cells were treated with 2.5, 5 and 10 μmol/L AMG-102, respectively. The proliferation activities of Hep-2 cells were detected by 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2H-tetrazolium bromide (MTT). The apoptotic rate of Hep-2 cells was detected by flow cytometry analysis and Hoechst staining. The mRNA expression levels of apoptosis-related genes were detected by real-time quantitative polymerase Chain reaction (RT-qPCR), and the protein expressions of c-Met/PI3K/AKT pathway were detected by western blot. Results: Compared with the control group, the proliferation rates of Hep-2 cells treated with 2.5, 5 and 10 μmol/L AMG-102 for 24 hours were (89.8±1.1)%, (79.8±1.0)% and (69.1±1.2)%, respectively; for 48 hours were (76.8±2.0)%, (60.2±1.1)% and (49.8±1.2)%, respectively; for 72 hours were (50.1±2.0)%, (41.5±1.1)% and (33.6±1.0), respectively, with significant differences (all P<0.05). The apoptotic rates of Hep-2 cells treated with 2.5, 5 and 10 μmol/L AMG-102 for 48 hours were (16.09±1.53)%, (27.51±2.02)% and (36.57±1.42)%, respectively, which were significantly higher than (3.62±0.10) % in the control group (all P<0.05). After treated with 2.5, 5 and 10 μmol/L AMG-102 for 48 hours, the relative expression levels of Bcl-2 mRNA in Hep-2 cells were 0.58±0.13, 0.38±0.12 and 0.20±0.13, respectively; the relative protein expression of p-Met were 80.0±3.8, 50.6±4.2 and 28.5±1.3, respectively; the relative protein expression of p-PI3K were 87.1±0.9, 54.2±1.2 and 21.0±1.2, respectively; the relative protein expression of p-AKT were 98.7±5.6, 56.9±3.2 and 32.2±4.3, respectively; which were significantly lower than those in the control group (all P<0.05). The relative expression levels of Bax mRNA were 1.78±0.13, 2.37±0.14 and 3.05±0.13, respectively, and the relative expression levels of caspase-3 mRNA were 1.98±0.14, 2.47±0.14 and 3.15±0.13, respectively, which were significantly higher than those in the control group (all P<0.05). Conclusion c-Met inhibitor AMG-102 could inhibit the proliferation and induce apoptosis of laryngeal squamous carcinoma Hep-2 cells by regulating the c-Met/PI3K/Akt pathway.

Chloroquine (CQ) phosphate has been suggested to be clinically effective in the treatment of coronavirus disease 2019 (COVID-19). To develop a physiologically-based pharmacokinetic (PBPK) model for predicting tissue distribution of CQ and apply it to optimize dosage regimens, a PBPK model, with parameterization of drug distribution extrapolated from animal data, was developed to predict human tissue distribution of CQ. The physiological characteristics of time-dependent accumulation was mimicked through an active transport mechanism. Several dosing regimens were proposed based on PBPK simulation combined with known clinical exposure-response relationships. The model was also validated by clinical data from Chinese patients with COVID-19. The novel PBPK model allows in-depth description of the pharmacokinetics of CQ in several key organs (lung, heart, liver, and kidney), and was applied to design dosing strategies in patients with acute COVID-19 (Day 1: 750 mg BID, Days 2-5: 500 mg BID, CQ phosphate), patients with moderate COVID-19 (Day 1: 750 mg and 500 mg, Days 2-3: 500 mg BID, Days 4-5: 250 mg BID, CQ phosphate), and other vulnerable populations (.., renal and hepatic impairment and elderly patients, Days 1-5: 250 mg BID, CQ phosphate). A PBPK model of CQ was successfully developed to optimize dosage regimens for patients with COVID-19.

Methods:Clinical data of 187 patients with left-sided hepatolithiasis and underwent laparoscopically or open left-sided hepatectomy from October 2014 to October 2019 at the Second Affiliated Hospital of Anhui Medical University were retrospectively analyzed in this propensity score matching (PSM) study and were matched in terms of age, sex, body mass index, liver function, ASA score, comorbidities, history of biliary surgery, and smoking history on the ratio of 1∶1.There were 47 cases in each group and the mean age were (54.7±12.3)years old(range:34 to 75 years old) and (53.2±12.6) years old (range: 34 to 75 years old) in open and laparoscopically group respectively. The data of operation time, intraoperative blood loss, postoperative hospital-stay, complication rate, biliary fistula rate, stone clearance rate, and stone recurrence rate were compared. The quantitative data were compared using t-test or rank-sum test. Count data were analyzed with χ 2 test or Fisher test. Results:No significant difference was observed in the clinical characteristics of included 94 patients in this study(all P>0.05).The length of the postoperative hospital-stay after OLH was significantly higher than that in the LLH group((10.8±3.1) days vs.(8.5±2.2)days, t=4.085, P=0.000). LLR significantly decreased the incidence of postoperative biliary fistula compared with the OLH (6.3% vs.21.2%, χ 2=4.374, P=0.036) and the rates of postoperative complications in the OLH group was significantly higher than that in the LLH group (48.9% vs.27.6%, χ 2=4.502, P=0.034). Moreover, the stone recurrence rates in the LLH group was significantly lower than that after OLR (4.2% vs. 17.0%, χ 2=4.029, P=0.045). OLH (95 % CI: 1.55 to 10.75, P=0.004) and postoperative complications (95 % CI: 1.29 to 9.52, P=0.013) were independent risk factors for prolonged hospital stay. OLH (95 % CI: 1.428 to 44.080, P=0.018) and residual stones (95 % CI: 1.580 to 62.379, P=0.014) were independent risk factors for the occurrence of postoperative biliary fistula. Biliary fistula (95 % CI: 1.078 to 24.517, P=0.040) was an independent risk factor for the recurrence of stones. Conclusion:Compared with OLH, LLH is safe and effective for the treatment of the primary left-sided hepatolithiasis with the clinical benefits of shorter hospital stay, fewer morbidity and biliary fistula occurrence, and lower stone recurrence rates.

Methods:Clinical data of 187 patients with left-sided hepatolithiasis and underwent laparoscopically or open left-sided hepatectomy from October 2014 to October 2019 at the Second Affiliated Hospital of Anhui Medical University were retrospectively analyzed in this propensity score matching (PSM) study and were matched in terms of age, sex, body mass index, liver function, ASA score, comorbidities, history of biliary surgery, and smoking history on the ratio of 1∶1.There were 47 cases in each group and the mean age were (54.7±12.3)years old(range:34 to 75 years old) and (53.2±12.6) years old (range: 34 to 75 years old) in open and laparoscopically group respectively. The data of operation time, intraoperative blood loss, postoperative hospital-stay, complication rate, biliary fistula rate, stone clearance rate, and stone recurrence rate were compared. The quantitative data were compared using t-test or rank-sum test. Count data were analyzed with χ 2 test or Fisher test. Results:No significant difference was observed in the clinical characteristics of included 94 patients in this study(all P>0.05).The length of the postoperative hospital-stay after OLH was significantly higher than that in the LLH group((10.8±3.1) days vs.(8.5±2.2)days, t=4.085, P=0.000). LLR significantly decreased the incidence of postoperative biliary fistula compared with the OLH (6.3% vs.21.2%, χ 2=4.374, P=0.036) and the rates of postoperative complications in the OLH group was significantly higher than that in the LLH group (48.9% vs.27.6%, χ 2=4.502, P=0.034). Moreover, the stone recurrence rates in the LLH group was significantly lower than that after OLR (4.2% vs. 17.0%, χ 2=4.029, P=0.045). OLH (95 % CI: 1.55 to 10.75, P=0.004) and postoperative complications (95 % CI: 1.29 to 9.52, P=0.013) were independent risk factors for prolonged hospital stay. OLH (95 % CI: 1.428 to 44.080, P=0.018) and residual stones (95 % CI: 1.580 to 62.379, P=0.014) were independent risk factors for the occurrence of postoperative biliary fistula. Biliary fistula (95 % CI: 1.078 to 24.517, P=0.040) was an independent risk factor for the recurrence of stones. Conclusion:Compared with OLH, LLH is safe and effective for the treatment of the primary left-sided hepatolithiasis with the clinical benefits of shorter hospital stay, fewer morbidity and biliary fistula occurrence, and lower stone recurrence rates.

Objective: To investigate the effect of c-Met inhibitor AMG-102 on proliferation and radiosensitivity in laryngeal squamous carcinoma cells. Methods: The effects of AMG-102 on proliferation and radiosensitivity of laryngeal squamous carcinoma cell lines Hep-2 and KBV200 were detected by 3-(4, 5-dimethy-2-thiazolyl)-2, 5-diphenyl-2H tetrazolium bromide (MTT) assay and colony formation assay, respectively. The apoptosis of Hep-2 and KBV200 cells was detected by flow cytometry. The expression levels of c-Met, phospho-Met (p-Met), cleaved caspase-3 and Akt/p-Akt, Erk/p-Erk were detected by Western blot. Specific small interfering RNA targeting c-Met or plasmid of c-Met were transfected into Hep-2 and KBV200 cells to investigate the cell sensitivity to AMG-102. Results: Compared with KBV200 cells, Hep-2 cells were more sensitive to AMG-102 with IC₅₀ of 14 and 9 μmol/L, respectively. The relative expression levels of c-Met and p-Met proteins in Hep-2 cells were 194.48±0.57 and 177.76±1.53, respectively, which were significantly higher than those in KBV200 cells (171.24±1.00 and 115.37±0.56, respectively, P<0.001 for both). Exogenous hepatocyte growth factor (HGF) was added to increase the expression level of p-Met protein in KBV200 cells. The results showed that AMG-102 significantly reduced the expression of p-Met in KBV200 cells treated with HGF (P<0.001). Compared with the dimethyl sulfoxide (DMSO) group, AMG-102 treatment combined with radiotherapy significantly increased the radiosensitivity of Hep-2 cells (SER=1.28, P<0.001). However, AMG-102 had little effect on the radiosensitivity of KBV200 cells (SER=1.18, P=0.002). Compared with the 4 Gy radiotherapy alone group and the 5 μmol/L of AMG-102 alone treatment group, the apoptosis rate of Hep-2 cells in the combined treatment group was significantly increased. Meanwhile, the expression level of cleaved caspase-3 protein was also markedly increased. However, there were no significant changes in the apoptotic rate and cleaved caspase-3 expression in each treatment group of KBV200 cells. Compared with DMSO treatment group, the expression levels of p-Met, p-Akt and p-Erk were significantly decreased in the 4 Gy radiotherapy group, 5 μmol/L of AMG-102 treatment group and combined treatment group of Hep-2 cells. And the levels of p-Met, p-Akt and p-Erk in the combined treatment group were significantly lower than those in the 4 Gy radiotherapy alone group and 5 μmol/L of AMG-102 treatment alone group. By contrast, in KBV200 cells, the expression of p-Met, p-Akt and p-Erk in each group was not changed. The relative expression of p-Met in Hep-2 cells before and after radiotherapy at 30 min, 1 h, 4 h, 8 h, 24 h were 99.89±0.61, 138.62±1.00, 163.07±5.00, 87.80±1.85, 90.67±0.65 and 94.09±1.41, respectively. The level of p-Met was slightly increased after radiotherapy at 30 min and 1 h (P<0.001 for all), whereas it was significantly decreased from 4 h to 24 h after radiotherapy (P<0.05 for all). By contrast, the expression of p-Met in KBV200 cells did not change with time after radiotherapy (P>0.05). The sensitivity of Hep-2 cells to AMG-102 was decreased after silencing of c-Met, while the sensitivity of KBV200 cells to AMG-102 was not significantly changed (P>0.05). Moreover, the radiosensitivity of Hep-2 cells in c-Met knockdown group had a slightly increasing trend (SER=1.07, P=0.068). After the treatment with 10 μmol/L of AMG-102, the proliferation rate of c-Met ectopically expressed KBV200 cells was 60.05%±3.23%, It was significantly lower than that of the blank control 90.08%±1.04% and siRNA negative control (90.12%±1.01%, P<0.001). The results suggested that the overexpression of c-Met in KBV200 cells increased the radiosensitivity to AMG-102, whereas depletion of c-Met resulted in resistance to AMG-102 in Hep-2 cells. Furthermore, the radiosensitivity of KBV200 cells that overexpressed c-Met showed a decreased trend (SER=0.7, P=0.005). Conclusions c-Met inhibitor AMG-102 has a significant inhibitory effect on the proliferation of c-Met overexpressing laryngeal squamous carcinoma cells, leading to increased radiosensitivity. It suggests that molecular targeted therapy against c-Met receptor is more effective in c-Met overexpressed subtype of laryngeal squamous cell carcinoma.

Objective To evaluate preoperative three dimensional(3D)reconstruction techniques in perioperative patients of hepatocellular carcinoma (HCC) undergoing hepatectomy.Methods Fifty-eight HCC patients who had undergone hepatectomy between 2015 and 2017 were enrolled.Twenty-three patients underwent hepatectomy based on preoperative 3D reconstruction techniques,while other thirty-five patients were without using it.Results No significant statistical difference was found in clincopathological parameters of patients preoperatively.The patients who underwent hepatectomy based on 3D reconstruction techniques had less operation time (Z =-2.213,P =0.028),hepatic inflow occlusion rate,time (x2 =3.966,P =0.046;Z =-2.371,P =0.018) and blood loss (Z =-2.140,P =0.032) during operation.Totally 23 postoperative complications occurred which were Clavien-Dindo classification grade Ⅰ or Ⅱ.More complications occurred in the not using 3D technique group (x2 =6.061,P =0.014).Conclusion Preoperative 3D reconstruction technique improves the perioperative prognosis of hepatectomy in patients with hepatocellular carcinoma.

Objective To investigate the effect of c?Met inhibitor AMG?102 on proliferation and radiosensitivity in laryngeal squamous carcinoma cells. Methods The effects of AMG?102 on proliferation and radiosensitivity of laryngeal squamous carcinoma cell lines Hep?2 and KBV200 were detected by 3?(4,5?dimethy?2?thiazolyl)?2, 5?diphenyl?2H tetrazolium bromide ( MTT ) assay and colony formation assay, respectively. The apoptosis of Hep?2 and KBV200 cells was detected by flow cytometry.The expression levels of c?Met, phospho?Met (p?Met), cleaved caspase?3 and Akt／p?Akt, Erk／p?Erk were detected by Western blot. Specific small interfering RNA targeting c?Met or plasmid of c?Met were transfected into Hep?2 and KBV200 cells to investigate the cell sensitivity to AMG?102. Results Compared with KBV200 cells, Hep?2 cells were more sensitive to AMG?102 with IC50 of 14 and 9 μmol／L, respectively. The relative expression levels of c?Met and p?Met proteins in Hep?2 cells were 194.48±0.57 and 177.76±1.53, respectively, which were significantly higher than those in KBV200 cells ( 171.24 ± 1.00 and 115.37 ± 0.56, respectively, P＜0.001 for both). Exogenous hepatocyte growth factor (HGF) was added to increase the expression level of p?Met protein in KBV200 cells.The results showed that AMG?102 significantly reduced the expression of p?Met in KBV200 cells treated with HGF ( P＜0.001). Compared with the dimethyl sulfoxide ( DMSO) group, AMG?102 treatment combined with radiotherapy significantly increased the radiosensitivity of Hep?2 cells ( SER＝1.28, P＜0.001). However, AMG?102 had little effect on the radiosensitivity of KBV200 cells (SER＝1.18, P＝0.002). Compared with the 4 Gy radiotherapy alone group and the 5 μmol／L of AMG?102 alone treatment group, the apoptosis rate of Hep?2 cells in the combined treatment group was significantly increased. Meanwhile, the expression level of cleaved caspase?3 protein was also markedly increased. However, there were no significant changes in the apoptotic rate and cleaved caspase?3 expression in each treatment group of KBV200 cells. Compared with DMSO treatment group, the expression levels of p?Met, p?Akt and p?Erk were significantly decreased in the 4 Gy radiotherapy group, 5 μmol／L of AMG?102 treatment group and combined treatment group of Hep?2 cells. And the levels of p?Met, p?Akt and p?Erk in the combined treatment group were significantly lower than those in the 4 Gy radiotherapy alone group and 5 μmol／L of AMG?102 treatment alone group. By contrast, in KBV200 cells, the expression of p?Met, p?Akt and p?Erk in each group was not changed. The relative expression of p?Met in Hep?2 cells before and after radiotherapy at 30 min, 1 h, 4 h, 8 h, 24 h were 99.89±0.61, 138.62±1.00, 163.07±5.00, 87.80±1.85, 90.67±0.65 and 94.09±1.41, respectively. The level of p?Met was slightly increased after radiotherapy at 30 min and 1 h (P＜0.001 for all), whereas it was significantly decreased from 4 h to 24 h after radiotherapy (P＜0.05 for all). By contrast, the expression of p?Met in KBV200 cells did not change with time after radiotherapy (P＞0.05). The sensitivity of Hep?2 cells to AMG?102 was decreased after silencing of c?Met, while the sensitivity of KBV200 cells to AMG?102 was not significantly changed ( P＞0.05). Moreover, the radiosensitivity of Hep?2 cells in c?Met knockdown group had a slightly increasing trend ( SER＝1.07, P＝0.068). After the treatment with 10 μmol／L of AMG?102, the proliferation rate of c?Met ectopically expressed KBV200 cells was 60.05%± 3.23%, It was significantly lower than that of the blank control 90.08%±1.04% and siRNA negative control (90.12%±1.01%, P＜0.001). The results suggested that the overexpression of c?Met in KBV200 cells increased the radiosensitivity to AMG?102, whereas depletion of c?Met resulted in resistance to AMG?102 in Hep?2 cells. Furthermore, the radiosensitivity of KBV200 cells that overexpressed c?Met showed a decreased trend (SER＝0.7, P＝0.005). Conclusions c?Met inhibitor AMG?102 has a significant inhibitory effect on the proliferation of c?Met overexpressing laryngeal squamous carcinoma cells, leading to increased radiosensitivity. It suggests that molecular targeted therapy against c?Met receptor is more effective in c?Met overexpressed subtype of laryngeal squamous cell carcinoma.