Objective:To explore the regulatory effect of glutathione S-transferase P1(GSTP1) on the radiosensitivity of mouse Lewis lung cancer (LLC) cells.Methods:GSTP1-shRNA lentivirus and negative control lentivirus were used to respectively infect the LLC cells, and stable transgenic strains were selected. Real-time PCR and Western blot were conducted to quantitatively measure the expression levels of GSTP1 mRNA and protein in the LLC cells to verify the knockdown effect. The cell counting kit-8(CCK-8) assay was used to detect cell viability after irradiation. The colony formation assay was utilized to assess the cell proliferation ability after irradiation. Flow cytometry was performed to assess the level of cell apoptosis after irradiation. The tumor-bearing mice were established and irradiated to detect the changes in the tumor volume after irradiation. TUNEL staining was employed to detect the level of tumor apoptosis after irradiation. Immunofluorescence was used to detect the number of CD 4+ CD 8+ T cells in the tumor after irradiation. Results:Real-time PCR and Western blot showed that after shRNA lentivirus interference, the expression levels of GSTP1 mRNA and protein were significantly down-regulated. Down-regulation of GSTP1 reduced cell viability and proliferation, and increased the rate of cell apoptosis after irradiation. The tumor volume of the tumor-bearing mice after irradiation in the GSTP1 knockdown group was significantly smaller than that in the NC group, whereas the tumor apoptosis rate was significantly higher and the number of infiltrating CD 4+ CD 8+ T cells in the tumor was remarkably higher compared with those in the control group. Conclusion:Knockdown of GSTP1 can significantly increase the radiosensitivity of LLC cells and enhance the infiltration of lymphocytes in tumor tissues.

Objective: To observe the changes of retinal microstructure in lamellar macular hole (LMH) after vitrectomy. Methods: A retrospective clinical observational study. Forty patients (41 eyes) with LMH and received vitrectomy in Ophthalmology Department of Peking University People’s Hospital from January 2014 to September 2018 were included in this study. Among them, 14 patients (15 eyes) were males and 26 patients (26 eyes) were females, with an average age of 67.8±8.6 years. There were 37 eyes with a lens and 4 eyes with an IOL. There were 29 eyes with LMH of tractional type, 7 eyes of degenerative type, and 5 eyes of mixed type. All patients underwent BCVA and OCT examinations. The BCVA examination was performed using the international standard visual acuity chart, which was converted into logMAR visual acuity. The average logMAR BCVA was 0.57±0.27; the mean macular retinal thickness (CRT) was 192.3±108.9 μm, the mean macular thickness (MRT) was 427.5±110.2 μm. Among the 29 eyes of tractional type, there were 17 eyes with retinal cavity, 8 eyes with macular retinoschisis, and 3 eyes with incomplete ellipsoid zone. Among the 7 eyes of degenerative type, there were 5 eyes with lamellar hole-associated epiretinal proliferation (LHEP), 5 eyes with retinal cavity, and 5 eyes with incomplete ellipsoid zone. Among the 5 eyes of mixed type, 2 eyes with LHEP, 1 eye with macular epiretinal membrane, and 4 eyes with incomplete ellipsoid zone. The average follow-up time after surgery was 12.8±5.2 months. Among them, 10 eyes were followed up for equal or greater than 24 months. After the surgery, the same equipment and method before the surgery were used for relevant examination. The changes of BCVA, CRT, and MRT before and after surgery were observed. Continuous variables were compared by t test. Results: At the last follow-up, the mean logMAR BCVA was 0.37±0.26. Compared with before surgery, the difference was statistically significant (t=5.98, P<0.01). The mean CRT and MRT were (245.2±90.8) and (347.0±46.7) μm, respectively. Compared with before surgery, the differences were statistically significant (t=-2.49, -5.24; P<0.05, <0.01). CRT and MRT changed greatly within 6 months after surgery, and then tended to be gentle. Among the 3 eyes with incomplete ellipsoid zone of tractional type before surgery, ellipsoid zone recovered in 2 eyes and partially recovered in 1 eye. Among the 17 eyes with retinal cavity and 8 eyes with macular retinoschisis before surgery, there were still 4 eyes with retinal cavity, but all the retinoschisis were disappeared. Among the 5 eyes with retinal cavity of degenerative type before surgery, there were still 2 eyes with retinal cavity and all the eyes with incomplete ellipsoid zone. Among 10 eyes with a follow-up time of equal or greater than 24 months, the macular ganglion cell complex partially atrophied in 6 eyes, and the nerve fiber layer separated in 2 eyes. There was no full-thickness macular hole after surgery. Conclusion For most LMH patients, vitrectomy can effectively improve the visual acuity and promote the recovery of retinal microstructure.

Objective:To investigate the clinical features and multimodal imaging features of eyes with perifoveal exudative vascular anomalous complex (PEVAC).Methods:A retrospective case study. From February 2014 to November 2020, 7 eyes of 7 patients with PEVAC diagnosed by ophthalmology examination in Department of Ophthalmologyof Peking University People's Hospital were included in this study. There were 6 males and 1 female. The age was 60.1±9.1 years. All were monocular. The chief complaints of visual deformation and vision loss were 3 and 1 cases, respectively. All patients underwent best corrected visual acuity (BCVA), fundus color photography, optical coherence tomography (OCT), fundus fluorescein angiography (FFA). BCVA examination was performed using the standard logarithmic visual acuity chart, which was converted to logarithm of the minimum angle of resolution (logMAR) visual acuity. OCT angiography (OCTA) and indocyanine green angiography (ICGA) were performed in 4 and 2 eyes, respectively. Three eyes were treated with intravitreal injection of anti-vascular endothelial growth factor (VEGF) combined with local laser photocoagulation. Two eyes were treated with laser photocoagulation alone. The follow-up time was 16.7±19.1 months. During follow-up, relevant examinations were performed with the same equipment and methods as at the initial diagnosis. The multimodal imaging characteristics and treatment response of the affected eyes were observed.Results:The baseline logMAR BCVA was 0.33±0.19 (0.20-0.80). All eyes showed isolated hemangiomatous lesions in the macular fovea with rigid retinal exudation, and 2 adjacent isolated hemangiomatous lesions were observed in 1 eye. FFA and ICGA examination showed that all eyes with macular hemangiomatous lesions showed clear boundary and strong fluorescence in the early stage. No other retinal or choroidal vascular abnormalities were observed. On OCT examination, circular lumen-like structures with strong reflective wall near the fovea were observed in the macular region of all eyes, accompanied by intraretinal cystic lumen. The macular central retinal thickness (CMT) was 326±125 (207-479) μm. In the four eyes examined by OCTA, blood flow signals were observed in the circular lumenoid structures with strong reflective walls adjacent to the fovea. Blood flow signals were observed in the superficial capillary layer (SCP) and deep capillary layer (DCP) of the retina in 3 eyes. SCP showed blood flow signal in 1 eye. In 4 eyes treated with intravitreal injection of anti-VEGF drugs, there was no significant improvement in the intraretinal capsule space after treatment. Subretinal fluid absorption, retinal cystoid edema persisted, and rigid exudation decreased in 1 eye. CMT decreased and BCVA increased in 5 eyes treated with laser photocoagulation or laser photocoagulation alone. At last follow-up, logMAR BCVA was 0.16±0.06 (0.10-0.20) and CMT was 212±34 (154-252) μm. Compared with baseline, the difference of BCVA was statistically significant ( t=2.661, P=0.037). Conclusions:The fundus of PEVAC patients is characterized by solitary or multiple solitary hemangiomatous lesions in the macular fovea. Round lumenoid structures with strong reflective walls, with or without intraretinal cystic lumen, rigid exudate, and subretinal fluid, in which blood flow signals can be seen in OCT.

Objective:To assess the efficacy and safety of the treat-and-extend (TAE) regimen and pro re nata (PRN) regimen of intravitreal conbercept in polypoidal choroidal vasculopathy (PCV) patients.Methods:A non-randomized controlled study was performed.Ninety-one patients (91 eyes) diagnosed with treatment-na?ve PCV from October 2016 to January 2019 at Department of Ophthalmology, Peking University People's Hospital were enrolled.All the patients received the intravitreal injection of 0.5 mg conbercept.After the initial treatment, the patients were divided into 3+ PRN group and 3+ TAE group according to their willingness.The follow-up time was one year.All the eyes underwent visual acuity test with ETDRS chart, optical coherence tomography (OCT) examination, fundus fluorescein angiography (FFA) and indocyanine green angiography (ICGA). Best corrected visual acuity (BCVA), central retinal thickness (CRT), maximum retinal thickness (MRT), pigment epithelium detachment (PED) height, the number and area of polypoidal lesions, the area of retinal hemorrhage and the area of branching vascular network (BVN) were recorded.Treatment interval and injection frequencies during the one-year follow-up were compared between the two groups.This study adhered to the Declaration of Helsinki.The study protocol was approved by Peking University People's Hospital (No.2020PHB250-01). Written informed consent was obtained from each patient.Results:One-year after treatment, the BCVA improvement in the 3+ PRN group and 3+ TAE group was 5.0(-2.0, 15.0) and 6.0(-1.0, 14.0) letters, respectively, showing no significant difference ( Z=-0.352, P=0.725). No significant differences were found in CRT, MRT and PED height between the two groups ( Z=-0.145, -0.529, -0.985, all at P>0.05). There was no significant difference in polypoidal lesions number, polypoidal lesions area, the number of eyes with different degrees of polyp regression, BVN area and retinal hemorrhage area between the two groups ( Z=-0.502, -0.300, -0.047, -0.265, -1.243, all at P>0.05). After the one-year follow-up, the mean injection frequency of 3+ PRN group was (7.6±0.9) times, which was lower than (8.4±2.0) times of 3+ TAE group, showing a significant difference ( t=2.432, P=0.019). The mean follow-up frequency was (11.3±1.5) times of 3+ PRN group, which was significantly higher than (10.1±1.7) times of 3+ TAE group ( t=3.403, P=0.001). For the 3+ TAE group, 17.1%(6/35) of patients achieved an extension interval of 12 weeks after the first 3 doses, and 48.5%(17/35) of patients achieved an extension interval of 8 weeks or more, with a mean maximum extension interval of (9.5±2.0) weeks.During the follow-up, 10 patients in 3+ PRN group and 8 patients in 3+ TAE group received photodynamic therapy as a rescue treatment. Conclusions:The 3+ PRN and 3+ TAE regimens of intravitreal injection of conbercept combined with photodynamic therapy as a rescue treatment have similar efficacy in visual and anatomical outcomes for PCV patients.3+ TAE regimen has a higher treatment frequency and fewer follow-up visits.

Poria is an important medicine for inducing diuresis to drain dampness from the middle energizer. However, the specific effective components and the potential mechanism of Poria remain largely unknown. To identify the effective components and the mechanism of Poria water extract (PWE) to treat dampness stagnancy due to spleen deficiency syndrome (DSSD), a rat model of DSSD was established through weight-loaded forced swimming, intragastric ice-water stimulation, humid living environment, and alternate-day fasting for 21 days. After 14 days of treatment with PWE, the results indicated that PWE increased fecal moisture percentage, urine output, D-xylose level and weight; amylase, albumin, and total protein levels; and the swimming time of rats with DSSD to different extents. Eleven highly related components were screened out using the spectrum-effect relationship and LC-MS. Mechanistic studies revealed that PWE significantly increased the expression of serum motilin (MTL), gastrin (GAS), ADCY5/6, p-PKAα/β/γ cat, and phosphorylated cAMP-response element binding protein in the stomach, and AQP3 expression in the colon. Moreover, it decreased the levels of serum ADH, the expression of AQP3 and AQP4 in the stomach, AQP1 and AQP3 in the duodenum, and AQP4 in the colon. PWE induced diuresis to drain dampness in rats with DSSD. Eleven main effective components were identified in PWE. They exerted therapeutic effect by regulating the AC-cAMP-AQP signaling pathway in the stomach, MTL and GAS levels in the serum, AQP1 and AQP3 expression in the duodenum, and AQP3 and AQP4 expression in the colon.
Animals ; Rats ; Poria ; Spleen ; Albumins ; Chromatography, Liquid ; Cyclic AMP Response Element-Binding Protein

OBJECTIVE：To establis h the UPLC fingerprint of Poria co cos aqueous extract ，and to investigate its relationship with sedative and hypnotic effect. METHODS ：Ten batches of P. cocos from different areas were extracted with water to obtain the aqueous extract. UPLC method was adopted. The determination was performed on Waters HSS-C 18 column with mobile phase consisted of 0.1％ phosphoric acid solution-acetonitrile-methanol （gradient elution ） at the flow rate of 0.4-0.2 mL/min. The detection wavelengths were set at 210 and 242 nm. The column temperature was 40 ℃，and sample size was 2 μL. The fingerprints of 10 batches of P. cocos aqueous extracts were established by using the Similarity Evaluation System of TCM Fingerprint （2012A version），and the common peaks were identified. The sedative and hypnotic effects of 10 batches of P. cocos aqueous extracts from different areas under the synergistic action of pentobarbital sodium were investigated by taking the sleeping rate ，sleep latency and sleep duration of mice as the single efficacy index. After data transformation of single efficacy index and total efficacy （single indexes calculated by analytic hierarchy process ），grey correlation analysis was used to analyze the correlation between the common peaks in fingerprint of P. cocos aqueous extract and the single efficacy index and total efficacy. RESULTS ：There were 24 common peaks in 10 batches of aqueous extract of P. cocos ， and 11 components were identified ， i.e. 16 α-hydroxydehydrotrametenolic acid （peak 6），16α-hydroxytrametendic acid （peak 7），poricoic acid B （peak 9），dehydrotumulosic acid（peak 10），poricoic acid A （peak 12），polyporenic acid C （peak 15），3-O-acetyl-16α-hydroxydehydrotrametenolic acid （peak 17），dehydropachymic acid （peak 20），pachymic acid （peak 21），dehydrotrametenolic acid （peak 22），dehydroeburicoic acid （peak 24）. Grey correlation analysis showed ，the correlation between 24 peaks and sleep duration was greater than 0.6（0.611 5- 0.811 8）；the correlation between 24 peaks and sleep latency was greater than 0.6（0.605 9-0.790 4），except for peaks 14，24 and 2；the correlation of 24 peaks between sleeping rate was greater than 0.6（0.606 4-0.721 6），except for peaks 23，19，17 and 5； the correlation of 24 peaks between total efficacy was greater than 0.6（0.619 0-0.781 2），except for peaks 2，5，19. The top 10 chromatographic peaks related to the total efficacy were peak 15（polyporenic acid C ），peak 16，peak 8，peak 11，peak 12 （poricoic acid A ）， peak 1， peak 7 （16 α-hydroxytrametendicacid）， peak 3， peak 9 （poricoic acid B ） and peak 20 （dehydropachymic acid ）. CONCLUSIONS ：UPLC fingerprint of P. cocos aqueous extract was established and 11 components were identified. Ten components such as polyporus acid C are closely related to the total efficacy of sedation and hypnosis ，which preliminarily reveal the material basis of the sedative and hypnotic effect of P. cocos .

OBJECTIVE：To study the repa ir，anti-inflammatory and analgesic effects of Compound crocodile oil burn ointment on superficial second-degree burned skin. METHODS ：The heated weight was attached to the right depilated skin of guinea pigs for 4 s to induce the model of superficial second-degree burn. After modeling ，guinea pigs were randomly divided into model group ， Jingwanhong ointment group （positive control ），formula Ⅰ，Ⅱ and Ⅲ groups of Compound crocodile oil burn ointment （volume fraction 1.5％，3％，4.5％，hereinafter），with 8 guinea pigs in each group. Except for model group ，other groups were smeared with 0.7 g/guinea pigs twice a day for 14 consecutive days. The wound healing was recorded every day ，the healing rate of wound was calculated. HE staining was used to observe the histopathological changes of the wound. The serum levels of EGF ，VEGF， SOD，MDA，TNF-α and IL-1 were detected by ELISA. Eighty Kunming mice were divided into 2 groups，and then sub-grouped into model group ，Jingwanhong ointment group （positive control ），formula Ⅰ and Ⅲ groups of Compound crocodile oil burn ointment，with 10 mice in each group. Then xylene auricle swelling method and acetic acid writhing method were used to investigate the anti-inflammatory and analgesic effects of Compound crocodile oil burn ointment. RESULTS ：In the burn repair experiment，after intervention of Compound crocodile oil burn ointment ，the wound area of guinea pigs gradually decreased ，and on the 14th day ，the wound had healed greatly ，and the wound healing rate increased significantly （P＜0.01）；serum levels of EGF and SOD were increased significantly （P＜0.01），while the levels of VEGF ，MDA，TNF-α and IL-1 were decreased significantly（P＜0.05 or P＜0.01）. The thick new epidermal layer was found in wound tissue ，and the connective tissue and neovascularization in the dermis increased significantly. In the anti-inflammatory and analgesic experiment ，after intervention of Compound crocodile oil burn ointment ，the degree of ear swelling and the times of writhing decreased significantly （P＜0.05 or P＜0.01）. CONCLUSIONS ：Compound crocodile oil burn ointment shows good skin repair ，anti-inflammatory and analgesic efficacy；the mechanism may be associated with increasing the serum levels of EGF and SOD and reducing the levels of VEGF ， MDA，TNF-α，IL-1.