Objective To elucidate the molecular mechanisms underlying the effects of Kanggan Mixture(KGM)on key targets in rats with acute lung injury,network pharmacology and in vivo micro-CT experiments were employed.Methods Network pharmacology was utilized to forecast the target genes and principal pathways involved in the intervention of KGM in acute lung injury(ALI).Lipopolysaccharide(LPS)-induced ALI rat models were utilized,and micro-computed tomography(micro-CT)was employed to evaluate the extent of lung injury in vivo.Experiments were conducted to verify the intervention mechanism of KGM on ALI rats.Results The findings revealed that 190 chemical constituents were identified from KGM,and 579 potential targets and 204 pathways associated with KGM's impact on ALI were predicted.The principal components of KGM,such as quercetin,luteolin,kaempferol,betulin,and lupenone,exhibit anti-viral,anti-inflammatory,and immunomodulatory properties by targeting TP53,AKT1,SRC,EP300,and STAT3,and modulating the FoxO signaling pathway,TNF signaling pathway,PI3K-Akt signaling pathway,and MAPK signaling pathway,demonstrating an influence on acute lung injury.Micro-CT results suggest that KGM can improve lung texture enhancement and lung injury in ALI rats,with an increase in end-expiratory lung volume(inspiratory phase-expiratory phase).The HE and W/D ratio results indicate that KGM can improve lung tissue injury and reduce the lung tissue wet/dry weight ratio(P<0.01).Blood cell analysis results show that the anti-inflammatory agent can decrease the WBC(white blood cell count)and N%(neutrophil percentage)in ALI rats'blood(P<0.01),and increase lymphocytes(P<0.05).Real-time quantitative PCR,WES,and immunohistochemistry results suggest that KGM can decrease the mRNA expression,protein distribution,and protein expression levels of TP53,AKT1,SRC,EP300,and STAT3 in lung tissue of ALI rats(P<0.05).Conclusion KGM has a certain intervention effect on acute lung injury,mainly achieved through the core targets STAT3,EP300,SRC,AKT1,and TP53.

Objective To investigate the effects of galectin-3(Gal3)on autophagy in high glucose-induced human telomerase reverse transcriptase-immortalized retinal pigment epithelial(hTERT-RPE)cells.Methods The hTERT-RPE cells cultured in vitro were randomly divided into a normal group(group C),a high glucose group(group HG),a high glu-cose+si-NC group(group HG+si-NC)and a high glucose+si-Gal3 group(group HG+si-Gal3).Reverse transcrip-tion polymerase chain reaction(RT-PCR)was used to detect the relative mRNA expression levels of Gal3,intercellular ad-hesion molecule(ICAM)-1,interleukin(IL)-6 and tumor necrosis factor(TNF)-α in hTERT-RPE cells of each group.The expression levels of IL-1 and IL-6 in the supernatant of hTERT-RPE cells were measured by enzyme-linked immunosorbent assay(ELISA).The expression level of reactive oxygen species(ROS),the activity of superoxide dismutase(SOD)and the content of malondialdehyde(MDA)in hTERT-RPE cells were analyzed by dichlorofluorescin diacetate(DCFH-DA)staining,the colorimetric method and the microplate method,respectively.The protein expression levels of Gal3,the ratio of microtubule associated protein 1 light chain 3(LC3)type Ⅱ to type Ⅰ(LC3 Ⅱ/LC3 Ⅰ),Beclin 1,and autophagy-associat-ed 5(ATG5)and 7(ATG7)in hTERT-RPE cells from each group were detected by Western blot analysis.After the hTERT-RPE cells in each group were transfected with double fluorescent mRFP-GFP-LC3 plasmids alone and with RFP-LAMP1 and GFP-LC3 plasmids jointly,the changes of autophagy flow in hTERT-RPE cells were detected by laser confocal microscopy.Results Compared with the group C,the group HG showed an increase in the expression of Gal3,IL-1,IL-6,TNF-α,ICAM-1,P62,ROS and MDA content(all P＜0.05).However,the expression of LC3 Ⅱ/LC3 Ⅰ,Beclin1,ATG7,ATG5,the SOD activity,and the number of red(autolysosomes)and yellow fluorescence spots(autophagosomes)of the double fluo-rescent mRFP-GFP-LC3 plasmid were all lower in the group HG than those in the group C(all P＜0.05).There was no sig-nificant difference in the number of yellow fluorescent spots(autolysosomes)co-located by RFP-LAMP1 and GFP-LC3 plas-mids between groups C and HG(P＞0.05).There were no significant differences in the expression levels of the above-mentioned indexes in hTERT-RPE cells between groups HG and HG+si-NC(all P＞0.05).The expression levels of Gal3,IL-1,IL-6,TNF-α,ICAM-1,P62,ROS and MDA content in hTERT-RPE cells of the group HG+si-Gal3 were lower than those of the group HG+si-NC(all P＜0.05).Compared with those in the group HG+si-NC,the expression levels of LC3 Ⅱ/LC3 Ⅰ,Beclin1,ATG7,ATG5,SOD activity,and the number of red and yellow fluorescent spots of the double fluo-rescent mRFP-GFP-LC3 plasmid were increased in the group HG+si-Gal3(all P＜0.05).There was no significant differ-ence in the number of yellow fluorescent spots co-located by RFP-LAMP1 and GFP-LC3 plasmids between groups HG+si-GaL3 and HG+si-NC(P＞0.05).Conclusion Gal3 is significantly elevated in hTERT-RPE cells induced by high glu-cose,resulting in impaired autophagy flow.It produces a direct regulatory effect on the formation of autophagosomes,and plays a highly active role in oxidative damage and expression of inflammatory factors in hTERT-RPE cells induced by high glucose.

Objective To elucidate the molecular mechanisms underlying the effects of Kanggan Mixture(KGM)on key targets in rats with acute lung injury,network pharmacology and in vivo micro-CT experiments were employed.Methods Network pharmacology was utilized to forecast the target genes and principal pathways involved in the intervention of KGM in acute lung injury(ALI).Lipopolysaccharide(LPS)-induced ALI rat models were utilized,and micro-computed tomography(micro-CT)was employed to evaluate the extent of lung injury in vivo.Experiments were conducted to verify the intervention mechanism of KGM on ALI rats.Results The findings revealed that 190 chemical constituents were identified from KGM,and 579 potential targets and 204 pathways associated with KGM's impact on ALI were predicted.The principal components of KGM,such as quercetin,luteolin,kaempferol,betulin,and lupenone,exhibit anti-viral,anti-inflammatory,and immunomodulatory properties by targeting TP53,AKT1,SRC,EP300,and STAT3,and modulating the FoxO signaling pathway,TNF signaling pathway,PI3K-Akt signaling pathway,and MAPK signaling pathway,demonstrating an influence on acute lung injury.Micro-CT results suggest that KGM can improve lung texture enhancement and lung injury in ALI rats,with an increase in end-expiratory lung volume(inspiratory phase-expiratory phase).The HE and W/D ratio results indicate that KGM can improve lung tissue injury and reduce the lung tissue wet/dry weight ratio(P<0.01).Blood cell analysis results show that the anti-inflammatory agent can decrease the WBC(white blood cell count)and N%(neutrophil percentage)in ALI rats'blood(P<0.01),and increase lymphocytes(P<0.05).Real-time quantitative PCR,WES,and immunohistochemistry results suggest that KGM can decrease the mRNA expression,protein distribution,and protein expression levels of TP53,AKT1,SRC,EP300,and STAT3 in lung tissue of ALI rats(P<0.05).Conclusion KGM has a certain intervention effect on acute lung injury,mainly achieved through the core targets STAT3,EP300,SRC,AKT1,and TP53.

Objective To investigate the effects of galectin-3(Gal3)on autophagy in high glucose-induced human telomerase reverse transcriptase-immortalized retinal pigment epithelial(hTERT-RPE)cells.Methods The hTERT-RPE cells cultured in vitro were randomly divided into a normal group(group C),a high glucose group(group HG),a high glu-cose+si-NC group(group HG+si-NC)and a high glucose+si-Gal3 group(group HG+si-Gal3).Reverse transcrip-tion polymerase chain reaction(RT-PCR)was used to detect the relative mRNA expression levels of Gal3,intercellular ad-hesion molecule(ICAM)-1,interleukin(IL)-6 and tumor necrosis factor(TNF)-α in hTERT-RPE cells of each group.The expression levels of IL-1 and IL-6 in the supernatant of hTERT-RPE cells were measured by enzyme-linked immunosorbent assay(ELISA).The expression level of reactive oxygen species(ROS),the activity of superoxide dismutase(SOD)and the content of malondialdehyde(MDA)in hTERT-RPE cells were analyzed by dichlorofluorescin diacetate(DCFH-DA)staining,the colorimetric method and the microplate method,respectively.The protein expression levels of Gal3,the ratio of microtubule associated protein 1 light chain 3(LC3)type Ⅱ to type Ⅰ(LC3 Ⅱ/LC3 Ⅰ),Beclin 1,and autophagy-associat-ed 5(ATG5)and 7(ATG7)in hTERT-RPE cells from each group were detected by Western blot analysis.After the hTERT-RPE cells in each group were transfected with double fluorescent mRFP-GFP-LC3 plasmids alone and with RFP-LAMP1 and GFP-LC3 plasmids jointly,the changes of autophagy flow in hTERT-RPE cells were detected by laser confocal microscopy.Results Compared with the group C,the group HG showed an increase in the expression of Gal3,IL-1,IL-6,TNF-α,ICAM-1,P62,ROS and MDA content(all P＜0.05).However,the expression of LC3 Ⅱ/LC3 Ⅰ,Beclin1,ATG7,ATG5,the SOD activity,and the number of red(autolysosomes)and yellow fluorescence spots(autophagosomes)of the double fluo-rescent mRFP-GFP-LC3 plasmid were all lower in the group HG than those in the group C(all P＜0.05).There was no sig-nificant difference in the number of yellow fluorescent spots(autolysosomes)co-located by RFP-LAMP1 and GFP-LC3 plas-mids between groups C and HG(P＞0.05).There were no significant differences in the expression levels of the above-mentioned indexes in hTERT-RPE cells between groups HG and HG+si-NC(all P＞0.05).The expression levels of Gal3,IL-1,IL-6,TNF-α,ICAM-1,P62,ROS and MDA content in hTERT-RPE cells of the group HG+si-Gal3 were lower than those of the group HG+si-NC(all P＜0.05).Compared with those in the group HG+si-NC,the expression levels of LC3 Ⅱ/LC3 Ⅰ,Beclin1,ATG7,ATG5,SOD activity,and the number of red and yellow fluorescent spots of the double fluo-rescent mRFP-GFP-LC3 plasmid were increased in the group HG+si-Gal3(all P＜0.05).There was no significant differ-ence in the number of yellow fluorescent spots co-located by RFP-LAMP1 and GFP-LC3 plasmids between groups HG+si-GaL3 and HG+si-NC(P＞0.05).Conclusion Gal3 is significantly elevated in hTERT-RPE cells induced by high glu-cose,resulting in impaired autophagy flow.It produces a direct regulatory effect on the formation of autophagosomes,and plays a highly active role in oxidative damage and expression of inflammatory factors in hTERT-RPE cells induced by high glucose.

Objective:To establish the fingerprint of standard decoction of Citri Reticulatae Pericarpium and evaluate its quality. Method:According to the preparation conditions of the standard decoction,15 batches of standard decoction of Citri Reticulatae Pericarpium were prepared.HPLC was employed to determine the content of hesperidin in this standard decoction.Ultraviolet spectroscopy(UV) and infrared spectroscopy(IR) were used to establish the fingerprint of standard decoction of Citri Reticulatae Pericarpium.The correlation coefficient method and double index sequence analysis method were used to compare and analyze the spectra of different batches of this standard decoction. Result:The content of hesperidin in 15 batches of this standard decoction were 0.82%-2.60%,and the measured value of dry extract rate was 32.02%-46.11%.Compared with ultraviolet and infrared control fingerprint,the fingerprint similarities of the standard decoction of each batch were > 0.897 and > 0.942,respectively.The double index analysis results showed that the common peak ratio was more than 62.50%,variation peak ratio was less than 46.67%. Conclusion:The quality evaluation method established in this study can be used for systematic evaluation of standard decoction of Citri Reticulatae Pericarpium,and it can provide theoretical reference for the formulation of quality standard of Citri Reticulatae Pericarpium dispensing granules and other related preparations.

Objective To fabricate an antibacterial controlled drug delivery system with PEG-hydrogel and gentamicin-loaded-CSt on titanium surface,and to investigate its surface characteristics,swelling behavior,drug release behavior in vitro,antiinfection performance in vivo,and tissue biocompatibility.Methods Cross-linked starch (CSt) was synthesized first and then CSt was loaded with gentamicin (GEN) as a carrier (GEN@CSt),then 4-arm-polyethylene glycol (PEG) was added to it which was mixed by ultrasound.The surface of titanium (Ti) was covered with a layer of poly dopamine (PDA).The drug-loaded hydrogel was fixed to the titanium surface,subsequently capped by poly lactic-co-glycolic acid (PLGA) membranes,and then the Ti-PDA-PEG (GEN@CSt)-PLGA composite coating was fabricated finally.Surface morphology of the system was observed,while the swelling behavior was characterized;release behavior of the composite coating was detected;the bacteriostatic experiments were carried out with staphylococcus aureus (SAU),staphylococcus epidermidis (SEP) and escherichia coli (ECO) in vitro.The animal models of infected bone defect was established in 36 New Zealand white rabbits.These animals were randomly divided into three groups.Group 1 animals were implanted with drug-loaded composite coatings.Group 2 animals were implanted with drug-free composite coatings.Group 3 animals were implanted with bare titanium rods.The infection data were collected periodically to carry out antiinfection experiments in vivo.Another 12 rabbits were divided into the experimental group and the control group randomly.Biocompatibility of the materials was observed by histopathology after implantation of the corresponding materials into the femoral condyle.Results The composite coating adhered to the titanium surface firmly,presenting a smooth and translucent shape.The ratio of CSt/PEG affects swelling behavior varied,starch-free gels maintained an equilibrium swelling of 7.4,after the ratio reached 1 ∶ 1,the equilibrium swelling ratio remained at 3.0.In-vitro the release rate of the first 8 h was fast,and the cumulative release amount accounted for 83％ of the total in the first 7 days,lasting more than 13 d.In vitro antibacterial test,the average diameter of the inhibition ring was 3.6±0.13 cm (SAU),3.4±0.11 cm (SEP),3.7±0.10 cm (ECO).In-vivo anti-infection experiment,the infection situation of the group 1 was better than the control groups 2 and 3.The pathological results indicated that inflammatory reaction in the experimental group was basically the same as the control group.Conclusion The study successfully fabricated the antibacterial controlled drug delivery system with PEG-hydrogel and gentamicin-loaded-CSt on titanium surface.The system has a reasonable drug release behavior,and effectively inhibited the growth of bacteria in vivo and in vitro.It also has good biocompatibility to stand a promising strategy to improve the orthopedics anti-infection.

Objective To evaluate the control effect of comprehensive measures of schistosomiasis after its transmission in?terruption in Kaihua County,Zhejiang Province,so as to provide the references for further consolidation of schistosomiasis con?trol. Methods The data of Oncomelania hupensis snail survey and control,as well as environmental reform in Kaihua County were collected and analyzed from 1996 to 2015. Results From 1996 to 2015,totally 2 635 snail habitats and 102.75 hm2 area with snails were found,and 125.4 thousand snails were dissected and no one was schistosome infected. The accumulated snail control area was 4 932.98 hm2,and the area with snails was effectively reduced by the comprehensive control measures. Conclu?sion The schistosomiasis control effect could be consolidated by the comprehensive control measures emphasizing environmen?tal reconstruction,and the snail surveillance work still should be strengthened.

Objective To evaluate the effect of triamcinolone orbital injection combined with dexamethasone in treatment of the patients with the grade Ⅳ of thyriod associated ophthalmopathy. Methods NOSPEVS Ⅳ grade of 90 patients were randomly divided into three groups,Group A,orbital injection of triamcinolone combind with dexamethasone, Croup B,simple injection of triamcinolone,Group C,oral prednisone therapy for half a year. To review each record of visual acuity intraocular pressure palpebral fissure size, upper eyelid drop late exophthalmos eye movement and so on every 4 weeks and follow-up for one year. Results Group A and group B of exophthalmos and eye movement improved obviously,however,Group C with poor efficacy and more side effects,and Group A in 3 groups was the fastest with least number of injections and which improved eye symptons significantly with fewer complications. Conclusion Orbital injection of triamcinolone combined with dexamethasone could be more rapid and effective than a simple injection of triamcinolone or oral prednisone to improve symptoms and signs in thyroid ophthalmopathy.

Objective To evaluate the relationship of diurnal variation of antidiuretic hormone (ADH) with urinary output,serum osmolality and blood pressure in spinal cord injury (SCI) patients. MethodsThe study was prospective,random and contrastive. Twenty complete SCI patients (two females and 18 males,Complete SCI group) and ten healthy controls (two females and eight males,control group) were studied. Urinary output and osmolality in the day time (8:00-20:00) and at night (20:00-8:00) were recorded. Blood samples for the measurement of serum osmolality and ADH were drawn at 14:00 and 2:00. Results There was very significant difference in regard of urinary output between day time and night time in complete SCI Group and control Group ( P 0.05). However,ADH level increased in the healthy Group at night,with a very significant difference ( P

In a fusion of BABL/C murine spleen cells immunizated with human fetal thymocytes and P_3X_(3)Ag_(,3), myeloma cells, six monoclonal antibodies(McAb) were produced. They were termed HIT_1. HIT_2. HIT_3. HIT_4.HIT_(6-1) and HIT_(6-2), respectively. The specificity of these McAbs were analysed by indirect immunofluorescence technique and FACS.Results showed that they reacted with 80～90%thymocytes,but hardly with peripheral blood mononuclear cells and spleen cells in adults,and nonreactive with red blood cells, granulocytes and platelets, According to their reaction with the tonsil cells, we can divide these six McAbs into three groups: Groupl including HIT_1, HIT_2, and HIT_(?) McAbs reacted approximately with 1/3 tonsil cells; basically GroupⅡ including HIT_(6-1) and HIT_(6-2) McAbs gave negative reaction with tonsil cells; GroupⅢ McAb HIT_4 reacted with 15% tonsil cells, which suggested these were a heterogeneous group McAbs with different specificities. In comparision with OKT series of McAbs in thymus, peripheral blood and tonsil, HIT_(1-3) are similar to OKT_(10) and,HTT6-l and HIT_(6-2) are just like OKT_6,but HIT_4 seems to be a new McAb different frOm HIT_(1-3) and HIT_(6-1) HII_(6-2). The competitive binding assay showed that HIT_(6-1) and HIT_(6-2) labeled with FITC can be inhibited by unlabeled HIT_(6-1) and HIT_(6-2) each other and can also be blocked by OKT_6, suggesting further these antibodies recognized a same epitope on thymocytes. Cross reaction were also demonstrated on HIT_1, and HIT_2 but not on HIT_3, suggesting HIT_1 and HIT_2 recognized the same determinant and HIT_3 recognized another. So six antibodies are McAbs against T cell differentiation antigens.They are useful for research the differentiation of T cells and the classification of malignant lymphadenosis diseases.