ObjectiveTo investigate the mechanism through which miR‑21 improves chronic renal fibrosis in mice via targeted modulation of the phosphatase and tensin homolog （PTEN）/protein kinase B （AKT）/mammalian target of rapamycin （mTOR） pathway. MethodsThirty‑two chronic kidney disease model mice were randomly divided into four groups （n=8 each group）： model group， miR‑21 overexpression group， miR‑21 inhibition group， and miR‑21 inhibition + MK‑2206 group. Eight healthy mice were included as the control group. The miR‑21 overexpression， miR‑21 inhibition， and miR‑21 inhibition + MK‑2206 groups received tail‑vein injections of lentivirus （50 μL， 1×10⁸ TU per mouse） once weekly for three weeks. The control and model groups were injected with an equal volume of empty vector （LV‑NC）. The miR‑21 inhibition + MK‑2206 group additionally received gavage of the AKT/mTOR pathway inhibitor MK‑2206 （480 mg/kg） once weekly for three weeks. The expressions of miR‑21， 24 h urinary protein， serum creatinine （Scr）， blood urea nitrogen （BUN）， and renal tissue levels of collagen Ⅰ， collagen Ⅲ， α‑smooth muscle actin （α‑SMA）， and PTEN protein， as well as p‑AKT/AKT and p‑mTOR/mTOR ratios， were compared among groups. HE staining was used to observe pathological changes in renal tissue， and Masson staining was used to observe the degree of renal fibrosis. A dual‑luciferase assay was performed to verify the targeting relationship between miR‑21 and PTEN. ResultsCompared with the model group， miR‑21 expression in renal tissue increased in the miR‑21 overexpression group （P<0.05） and decreased in the miR‑21 inhibition group （P<0.05）. Compared with the model group， the miR‑21 overexpression group showed increased 24 h urinary protein， Scr， BUN， and renal tissue expression of collagen Ⅰ， collagen Ⅲ， and α‑SMA （all P<0.05）， while these indicators decreased in the miR‑21 inhibition group （P<0.05）. Compared with the miR‑21 inhibition group， the miR‑21 inhibition + MK‑2206 group exhibited lower 24‑h urinary protein， Scr， BUN， and renal tissue expression of Collagen Ⅰ， Collagen Ⅲ， and α‑SMA （all P<0.05）. Compared with the model group， the miR‑21 overexpression group showed decreased PTEN protein expression （P<0.05） and increased p‑AKT/AKT and p‑mTOR/mTOR ratios （P<0.05）， while the miR‑21 inhibition group showed increased PTEN expression （P<0.05） and decreased p‑AKT/AKT and p‑mTOR/mTOR ratios （P<0.05）. Compared with the miR‑21 inhibition group， the miR‑21 inhibition + MK‑2206 group had lower p‑AKT/AKT and p‑mTOR/mTOR ratios （P<0.05）， with no significant difference in PTEN protein expression. HE and Masson staining showed normal kidney structure and almost no fibrosis in the control group. The model group exhibited glomerular enlargement， capillary loop adhesion， and focal fibrosis. The miR-21 overexpression group showed severe destruction of glomerular structure， accompanied by extensive fibrosis and renal tubular atrophy. The pathological changes and degree of fibrosis were alleviated in the miR-21 inhibition group. The miR-21 inhibition + MK-2206 group showed only mild pathological changes and mild fibrosis， with the interstitium being largely normal. Compared with PTEN-WT + NC mimics 1， the relative luciferase activity in the PTEN-WT + miR-21 mimics group decreased （P<0.001）. There was no statistically significant difference in relative luciferase activity between PTEN-WT + NC mimics group and PTEN-MUT + miR-21 mimics group. ConclusionmiR‑21 may improve renal function indicators and alleviate renal fibrosis in chronic kidney disease mice via targeted modulation of PTEN and subsequently inhibiting the AKT/mTOR pathway.

IGF-1 is involved in the growth and development of mammals,but its role in sebaceous gland excretion,B16 cell proliferation,and migration in the skin has not been reported yet.This study aims to reveal the function of IGF-1 by detecting its expression in animal skin.Using nano-body screening and purification methods,IGF-1 nanobodies(IGF-1-VHH)were obtained.Using IGF-1-VHH as the primary antibody,Western blot and immunohistochemistry were used to detect the expression of IGF-1 in bovine skin and alpaca acne.IGF-1-VHH was added to melanoma B16 cell culture medium,and CCK-8 and scratch healing methods were used to detect the effects of IGF-1 nanobodies on B16 cell proliferation and migration,as well as their possible molecular mech-anisms.The results showed that the obtained IGF-1-VHH could be applied in immunohistochemis-try and Western blot detection methods,with strong IGF-1 positive expression signals in bovine sebaceous glands and alpaca acne.IGF-1-VHH binds to IGF-1 in cells and has a certain inhibitory effect on the proliferation and migration of B16 cells by regulating the expression of RAS,ERK,and RAF.In summary,IGF-1 maybe involved in the secretion and excretion of sebum in animal skin.IGF-1-VHH inhibits the proliferation and migration of B16 cells by binding to IGF-1,provi-ding a new theoretical basis for ensuring normal physiological function of the skin and clinical di-agnosis and treatment of melanoma.

IGF-1 is involved in the growth and development of mammals,but its role in sebaceous gland excretion,B16 cell proliferation,and migration in the skin has not been reported yet.This study aims to reveal the function of IGF-1 by detecting its expression in animal skin.Using nano-body screening and purification methods,IGF-1 nanobodies(IGF-1-VHH)were obtained.Using IGF-1-VHH as the primary antibody,Western blot and immunohistochemistry were used to detect the expression of IGF-1 in bovine skin and alpaca acne.IGF-1-VHH was added to melanoma B16 cell culture medium,and CCK-8 and scratch healing methods were used to detect the effects of IGF-1 nanobodies on B16 cell proliferation and migration,as well as their possible molecular mech-anisms.The results showed that the obtained IGF-1-VHH could be applied in immunohistochemis-try and Western blot detection methods,with strong IGF-1 positive expression signals in bovine sebaceous glands and alpaca acne.IGF-1-VHH binds to IGF-1 in cells and has a certain inhibitory effect on the proliferation and migration of B16 cells by regulating the expression of RAS,ERK,and RAF.In summary,IGF-1 maybe involved in the secretion and excretion of sebum in animal skin.IGF-1-VHH inhibits the proliferation and migration of B16 cells by binding to IGF-1,provi-ding a new theoretical basis for ensuring normal physiological function of the skin and clinical di-agnosis and treatment of melanoma.

Objective To compare the efficacy and safety of entecavir versus adefovir dipivoxil in the treatment of HBeAg positive chronic hepatitis B ( CHB) . Methods Ninety?six cases with HBeAg positive CHB were divided into ETV group and ADV group according to different medication. In addition to conventional treatment,ETV group received entecavir 0. 5 mg/d,ADV group received adefovir dipivoxil 10 mg/d. HBV DNA negative conversion rate,alanine aminotransferase ( ALT) recurrence rate and HBeAg negative conversion rate in 24 weeks,48 weeks and 96 weeks were compared as well as the adverse reactions and liver function in 96 weeks. Results HBV DNA negative conversion rates in ETV group were significantly higher than those in ADV group in 24 weeks,48 weeks and 96 weeks (24 weeks:64. 6％(31/48) vs. 41. 7％(20/48);48 weeks:83. 3％(40/48) vs. 52. 1％(25/48);96 weeks:97. 9％(47/48) vs. 62. 5％(30/48),χ2 =5. 06,10. 72,18. 96,P<0. 05) . ALT recurrence rates in ETV group were significantly higher than those in ADV group at 24 weeks,48 weeks ( 24weeks:77. 1％( 37/48 ) vs. 54. 2％( 26/48 );48weeks:85. 4％( 40/48 ) vs. 62. 5％( 30/48 ) ,χ2=5. 59,6. 54,P<0. 05). There was no significant difference in ALT complication rate at 96 week(χ2=0. 71,P>0. 05) . There was no significant difference in HBeAg negative conversion rate between the two groups through treatment(χ2=0. 07, 0. 22, 0. 44, P>0. 05 ) . After 96 weeks, ALT in both groups decreased significantly ( t =13. 56,11. 85,P<0. 05) ,while ALT in ETV group was significantly lower than that in ADV group ( ( 31. 8 ±8. 6) U/L vs. (38. 5±7. 5) U/L,t=4. 07,P<0. 05). AST in both groups decreased significantly(t=41. 27, 33. 68,P<0. 05),while AST in ETV group was significantly lower than that in ADV group ( (30. 3±6. 5) U/L vs.(37.6±7.1)U/L,t=5.25,P<0.05).TBIL in both groups decreased significantly(t=28.92,22.23,P<0. 05),while TBIL in ETV group was significantly lower than that in ADV group ( (13. 5±3. 3) μmol/L vs. (18. 7±3. 9) μmol/L,t=7. 05,P<0. 05). GGT in both groups decreased significantly (t=16. 99,13. 97,P<0.05),while GGT in ETV group was significantly lower than that in ADV group ( (35.6±10.4)U/L vs. (59. 7±12. 5)U/L,t=10. 27,P<0. 05). There was no significant difference in adverse reaction between the two groups (χ2=1. 96,P>0. 05) . Conclusion Entecavir has a higher rate of HBV DNA negative conversion rate, ALT recurrence rate and HBeAg negative conversion rate in the treatment of HBeAg positive CHB. It is an ideal antiviral drug.

Objective To explore the therapeutic effect of lipid emulsion on acute organophosphorus poisoning and its consequence of acute lung injury. Methods A total of 48 sealant - grade Sprague-Dawley (SD) rats were randomly divided into four groups A,B,C,D,namely saline control group,lipid emulsion control group,the conventional therapy group and lipid emulsion administration group. After dichlorvos (DDVP) 11 mg/kg was given by intra-peritoneal injection,if there was no loss of DDVP during the injection process,the model of poisoning was considered to be made successfully.Then the rat models in four groups were respectively treated:with normal saline (5 ml/kg) intravenous injection in group A,lipid emulsion (5ml/kg) intravenous injection in group B,atropine (5 mg/kg) and pralidoxime chloride (40 mg/kg) intramuscular injection in group C,and combined use of lipid emulsion (5 ml/kg) with atropine and pralidoxime chloride in group D after administration of DDVP by intra-peritoneal injection.The activity of cholinesterase (CHE) in blood was detected before and 0.5 h,2 h and 4 h after DDVP poisoning. The clinical manifestations,the survival of rats,the wet weight of rat' s lung and the pathological changes of the lung tissue were observed within following 24 h. The rates of survival and symptoms of rats were compared between paired groups by using the x2 test,and the mean values of biomarkers were compared paired groups by using t test. Results In groups A and B,the intensity of muscular fasciculation and salivation were more severe and appeared sooner after DDVP exposure in comparison with groups C and D leading to lower survival rates in group A and B. Compared with group C,the rate of 24 h survival was higher and the intensity of muscular fasciculation was weaker in group D ( P ＜ 0.05 ).In group A and group B,the 24-hour survival rates were 1/12 and 2/12,respectively ( P ＜ 0.05 ).The levels of CHE in blood significantly decreased after DDVP poisoning ( P ＜ 0.05 ).There was no significant difference in activity of CHE between group B and group A,and in groups C and D,the levels of CHE in blood were not significantly higher than that in the group B 0.5 h after DDVP poisoning ( P ＜ O.05 ).In groups C and D,the activity of CHE in blood was significantly higher compared with group A and B,and that in group D was higher compared with C,and that in group B was higher compared with A 2 and 4 hours after DDVP poisoning ( P ＜ 0.05 ).In groups C and D,the wet weight of rat lung was significantly lighter compared with groups A and B,and that in group D was lighter compared with C,and that in group B was lighter compared with A 24 h after DDVP poisoning P ＜ 0.05 ).The electron microscopic findings showed the combined use of lipid emulsion with atropine and pralidoxime chloride obviously lessened the lung histopathologic changes after DDVP poisoning.Conclusions The lipid emulsion combined with atropine and pralidoxime chloride can be beneficial to controlling the toxic symptoms,reduce the death rate,accelerate the resume of the activity of CHE in blood,and relieve the lung injury induced by acute organophosphorus poisoning.

Objective To discuss the effect of Montelukast (Mont) on MDA,SOD,W/D,TNF-α,IL-10 and NF-κBp65 in lung tissue of Wistar rats poisoned by paraquat (PQ) and also to observe the pathological changes of the lung tissue.Methods A total of 104 Wistar rats were divided into 3 groups in random (random number),namely PQ group (n =40),Mont group (n =40) and control group (n =24).PQ (20 mg/kg) was administered by intra-peritoneal route to rats of PQ group and Mont group and narcotics were used for 2 hours.Mont in dose of 50 mg/kg was administered intra-gastrically to rats of Mont group per day and saline instead were administered to PQ group and control group per day until they were sacrificed for experiment.Of both PQ group and Mont group,10 rats were sacrificed at each interval of 1,3,5 and 7 days respectively after modeling,whereas 6 rats of control group were sacrificed at each interval.The levels of MDA and SOD in lung tissue and W/D of lung tissue,the levels of serum TNF-α and IL-10 and the level of NF-κBp65 in lung tissue were determined.Further,the specimen of lung tissue was prepared for electron microscopy observation.Results The level of MDA in lung tissue of PQ group was (8.19 ± 0.53) nmol/mg prot,which was significantly higher than that of control group on the 7th day.The level of SOD in lung tissue of PQ group was (128.76 ± 10.18) U/mg prot,which was significantly lower than that of control group.In PQ group,the W/D of lung tissue (6.62 ±0.42),level of serum TNF-α (156.16 ± 11.13) pg/ml,level of IL-10 (43.63 ±4.44) pg/ml and level of NF-κBp65 in lung tissue (0.23 ±0.02) were significantly higher than those in control group (P ＜0.01).In Mont group on the 7th day,the level of serum TNF-α (129.99 ±13.13) pg/ml,level of serum IL-10 (34.28 ± 3.80) pg/ml and level of NF-κBp65 in lung tissue (0.20 ±0.02) were significantly lower than those in PQ group (P ＜ 0.01).In the PQ group,pathological changes of lung tissue under the light and electron microscopes were acute diffused lung injury manifested itself in hemorrhage,effusion and infiltration of inflammatory cells inside the alveolar space,and the necrosis and defluxion of Ⅰ type and Ⅱ type epithelia cells.The pathological changes in Mont group were localized with infiltration of scanty inflammatory cells,and Ⅰ type epithelia cells were intact and there was no obvious necrosis of Ⅱ type epithelia cells.Conclusions Mont has protective effects on acute lung injury caused by PQ poisoning in rats.