Objective To observe the expression of toll like receptor 4(TLR4) Signaling and the release of inflammation factors in rat tubular epithelial cell(NRK-52E) under high glucose condition after TLR4-siRNA transfection.Methods Three TLR4-siRNA sequences were designed and synthesized.The transfection efficiency was observed by fluorescence microscope after transfection,and the expression of TLR4 mRNA was detected by real time PCR.The most effective siRNA was selected to be used for forward experiments.After transfection for 24 h,cells were stimulated with 25 mmol/L glucose and/or 10-7 mmol/L Angiotension Ⅱ (Ang Ⅱ) for 12 h,24 h; cells without stimulation were as normal control.Real-time PCR was used to analyze TLR4 and myeloid differentiation factor 88 (MyD88) mRNA expression; Western blot was used to observe TLR4/MyD88 and NF-κB protein expression.ELISA assay was used to detect the concentration of monocyte chemoattractant protein-1 (MCP-1),interleukin-6(IL-6) in cell supernatant after cells were stimulated for 24 h.Results TLR4/ MyD88 mRNA and TLR4/MyD88/NF-κB protein were highly expressed under high glucose or Ang Ⅱ co -incubated NRK-52E(P ＜ 0.01),the MCP-1 and IL-6 levels were also increased markedly compared with normal control group (P ＜ 0.01).TLR4/MyD88 mRNA and TLR4/MyD88/NF-kB protein expressions were obviously inhibited in cells that were transfected with TLR4-siRNA compared with high glucose group(P ＜ 0.01),MCP-1 and IL-6 production decreased remarkably compared with high glucose or Ang Ⅱ co-stimulated group(P ＜ 0.01).Conclusions High glucose can lead to the activation of TLR4/ MyD88/NF-kB signaling and the secretion of inflammation factors in NRK-52E,Ang Ⅱ further augments these effects.The effect can be blocked efficiently by specific siRNA gene silence.TLR4 signaling plays a pivotal role in the innate-immune inflammatory reaction in NRK-52E.

Objective To investigate the effects of angiotensin Ⅱ (Ang Ⅱ) or high glucose on the toll-like receptor 4 (TLR4) expression,inflammatory cytokines and fibrotic factors in human tubular epithelial cells (HK-2),revealing the innate immune-related pathogenesis of diabetic nephropathy (DN) which may have clinical implications.Methods Three TLR4 siRNA sequences were designed and synthetized.After transfection,the most effective siRNA was selected to use for further expriments.The experiment consisted of 2 parts.Part 1:Cells were divided into three groups:normal-glucose group (NG,5.5mmol/L glucose),mannose group (M,5.5 mmol/L glucose + 19.5 mmol/L mannose),High-glucose group (HG,25 mmol/L glucose),preliminary validated the effects of high glucose and high osmotic pressure.Part 2:Cells were divided into seven groups:NG group,HG group,Ang Ⅱ group,Ang Ⅱ + negative group,HG+ negative group,Ang Ⅱ + siRNA group and HG+ siRNA group.Real time PCR was used to analyze the mRNA expression of TLR4,myeloid differentiation factor 88 (MyD88),heat shock protein 47 (HSP47).Western blotting was used to observe the protein expression of TLR4,MyD88,HSP47,NF-κB,type Ⅳ collagen (ColⅣ).ELISA was used to detect the expression of monocyte chemotactic protein-1 (MCP-1) and interleukin-6 (IL-6).Results Compared with NG group,TLR4,MyD88,HSP47 mRNA and TLR4,MyD88,NF-κB,ColⅣ,HSP47 protein were highly expressed under high glucose or Ang Ⅱconditions (P ＜ 0.01),and the expression levels of MCP-1 and IL-6 also increased significantly (P ＜ 0.01).Compared with HG or Ang Ⅱ group,the above indicators were obviously inhibited in the TLR4 siRNA groups (P＜0.01).Comparison between blank vector transfected groups and HG group as well as Ang Ⅱ group indicated no statistic significance (P ＞ 0.05).Conclusions Both Ang Ⅱ and high glucose stimulate TLR4 expression,which result in the up-regulation of inflammatory and fibrotic factors in HK-2.Specific target of TLR4 gene silencing can block the TLR4 pathway that is activated by high glucose and Ang Ⅱ,and thus reduce the inflammatory and fibtogenic factors' release.TLR4 signal is the common innate immune response pathway which induces the release of inflammatory and fibrotic factors in HK-2 under high glucose or high angiotension conditions.

Objective: To investigate the stamina,the role of antioxidant system in severe acute pancreatitis(SAP),and the effect of fish oil treatment in rats.Methods:Thirty-six male Sprague-Dawley rats(SD) were randomly divided into three groups,including SAP group(sodium chloride treatment ) (group NSG,n=12),fish oil treatment group (group FOG,n=12),and control group (group CG,n=12).SAP was induced by intergraded injection of 3.5% sodium tanrocholate to biliopancreatic duct of SD rats in group NSG and FOG.The group NSG rats were treated by subcutaneous injection of sodium chloride,while the group FOG rats were treated by subcutaneous injection of fish oil for 7 days.Then the improved tail suspension test was observed at the first,third,fifth,seventh day.12 rats in each group were respectively sacrificed after 7 day.The activity of serum antioxidant enzymes (MDA,GSH-PH) and the concentration of serum amylase were measured in each group,and the levels of threshold on the area and the total immobility time were measured in each group.The severity of pancreatitis was analyzed according to the histopathological morphology.Results: Compared to group NSG,the severity of pancreatitis was significantly decreased in group FOG.The activity of MDA was significantly increased in group NSG than that in group CG (P<0.01) ,while the activity of MDA in group FOG was decreased than that in group NSG (P<0.05) .The activity of GSH-PH was significantly decreased in group NSG than that in group CG(P<0.01),while the activity of GSH-PH was increased in group GOG than that in group NSG (P<0.05).THE level of threshold on the area was decreased in group NSG than that in group CG(P<0.01),while the level of threshold on the area was increased in group GOG than that in group NSG.(P<0.01) The total immobility time was significantly increased in group NSG than that in group CG(P<0.01),while the total immobility time was decreased in group GOG than that in group NSG(P<0.05).MDA was associated with the level of threshold on the area and the total immobility time.Conclusion: Fish oil has a positive effect on the activity of antioxidant system and behavior character in SAP rats.

Objective:To analyze the in-hospital mortality and influential factors of total hip arthroplasty (THR) in older adult patients.Methods:A total of 130 older adult patients subjected to THR in Department of Orthopedics, Huzhou Linghu People's Hospital between August 2019 and August 2021 were included in this study. In-hospital mortality was calculated. These patients were divided into death and survival groups according to whether they were dead or alive. Sex, age, smoking, drinking, disease type, complications, cardiovascular disease history, operation severity score, operative time, amount of intraoperative blood loss, postoperative osteoporosis treatment and postoperative rehabilitation training were compared between the two groups. The risk factors that influence in-hospital mortality were analyzed using logistic analysis method.Results:Among 130 patients, 9 patients died in Huzhou Linghu People's Hospital, with the mortality of 6.92%. Thus, there were 9 patients in the death group and 121 patients in the survival group. There were significant differences in age, smoking, complications, operation severity score and postoperative osteoporosis treatment between the two groups ( t = 1.70, χ2 = 5.48, χ2 = 4.09, t = 2.86, χ2 = 4.03, all P < 0.05). Multivariate logistic analysis showed that the age (≥ 85 years old), smoking (yes), complications (≥ 3), operation severity score (≥ 15 points) were the risk factors that influence in-hospital mortality. Postoperative osteoporosis treatment was the protective factor of THR. Conclusion:The in-hospital mortality of older adult patients after THR is high. Age ≥ 85 years old, smoking, complications ≥ 3, and operation severity score ≥ 15 are the risk factors of THR and may greatly affect the prognosis. More attention should be paid to older adult patients who have these risk factors.

Objective:To investigate the efficacy of a proximal femoral locking plate (LPFP) versus a proximal femoral anti-rotation intramedullary nail (PFNA) in the treatment of femoral intertrochanteric fractures in older adult patients. Methods:A total of 130 older adult patients with femoral intertrochanteric fractures who received treatment in Linghu People's Hospital of Huzhou from May 2017 to June 2020 were included in this study. They were randomly assigned to undergo treatment with either a PFNA (observation group, n = 65) or an LPFP (control group, n = 65). Intraoperative blood loss, incision length, operative time, and time to fracture healing were determined in each group. At 1, 3, and 6 months after surgery, the Harris hip score was used to evaluate hip joint recovery. Coxa vara, incision infection, and internal fixation loosening were compared between the two groups. Results:Intraoperative blood loss in the observation group was less than that in the control group [(189.26 ± 48.15) mL vs. (96.47 ± 40.21) mL, t = -11.93, P < 0.001]. Incision length, operative time, and time to fracture healing in the observation group were significantly shorter than those in the control group [(4.03 ± 1.48) cm vs. (12.16 ± 1.55) cm, (72.13 ± 28.75) minutes vs. (120.34 ± 29.01) minutes, (9.89 ± 1.52) weeks vs. (13.63 ± 1.74) weeks, t = -30.59, -9.52, -13.05, all P < 0.001]. At 1 month after surgery, there was no significant difference in Harris hip score between the two groups ( t = 1.28, P > 0.05). At 3 and 6 months after surgery, the Harris hip score gradually increased in the control and observation groups ( F = 13.44, 8.26, both P < 0.001). At 3 and 6 months after surgery, Harris hip scores in the observation group were significantly higher than those in the control group [(85.17 ± 4.29) points vs. (79.50 ± 4.12) points, (95.30 ± 1.04) points vs. (87.69 ± 1.25) points, t = 7.69, 37.73, both P < 0.001]. The incidence of complications in the observation group was significantly lower than that in the control group [1.54% (1/65) vs. 10.77% (7/65), χ2 = 4.80, P = 0.029). Conclusion:Compared with LPFP, PFNA can effectively reduce intraoperative blood loss in older adult patients with femoral intertrochanteric fractures, accelerate the progress of fracture healing, promote the recovery of the hip joint, and has fewer complications. Therefore, PFNA is worthy of popularization.

Objective To explore the feasibility of tubeless 2 μm laser vaporesection in treating pediatric ureter cysts by ureteroscopy.MethodsClinical data of 33 ureter cysts patients who received tubeless 2 μm laser vaporesections by ureteroscopy were reviewed. The median age of patients was 4 years with a range from 1 to 7 years. The operations were carried out by RevoLix 2 μm laser through ureteroscopy without ureter stents and catheters indwelling.ResultsAll operations were successfully performed. And no serious complications occurred after the operations.ConclusionsTubeless transurethral 2 μm laser treatment by ureteroscopy was a superior micro-invasive surgery method for pediatrics with ureter cysts, with advantages of little blood loss, high safety, convenient operation and infrequent complications.

Objective:To investigate the application value of magnetic resonance neurography (MRN) in pelvic fractures with lumbosacral plexus injury.Methods:From January 2014 to June 2020, data of 9 patients with pelvic fractures combined with lumbosacral plexus injury who were diagnosed with MRN and were performed surgical treatment were analyzed retrospectively. Among them, there were 8 males and 1 female, aged from 18 to 54 years, with an average age of 40.6±11.0 years. There were 2 fresh fractures within 3 weeks and 7 old pelvic fractures in 9 patients. According to AO classification, there were 6 cases of C1.3 type, 3 cases of C3 type. According to Denis classification of sacral fracture, there were 8 cases of Denis II type, 1 case of Denis III. Gibbons classification of nerve injury: 2 cases of grade II, 4 cases of grade III, 3 cases of grade IV; According to the criteria by the Nervous Injury Committee of British Medical Research Council (BMRC): 2 cases of M1, 3 cases of M2, 2 cases of M3, 2 cases of M4. Before the operation, the MRN technology was used to accurately locate the location and extent of the lumbosacral nerve injury, and nerve exploration release or internal fixation combined with nerve exploration release was employed for treatment.Results:All 9 patients underwent surgery successfully. The operation time ranged from 150 to 360 min, with an average of 217.8±63.8 min; intraoperative blood loss ranged from 200 to 1 100 ml, with an average of 388.9±293.4 ml. All patients were followed up for 21.6±19.3 months. All the fractures were healed, and the healing time was 12.7±2.2 months (range, 10-18 months). At the last follow-up, Gibbons classification of nerve injury: 3 cases of grade I, 5 cases of grade II, and 1 case of grade III. According to the criteria of BMRC: the muscle strength achieved M5 in 6 cases, M4 in 2 cases, M3 in 1 case. One patient developed a urinary system infection 2 months after discharge and recovered after treatment.Conclusion:MRN technique can assist the surgeon to fully understand the compression and traction damage of the lumbosacral nerve by the fracture before the operation, and make clear qualitative and localized diagnosis of the nerve damage, making the operation more precise and minimally invasive, which is a supplement to the traditional diagnosis method of lumbosacral plexus injury.

Objective:To explore the clinical diagnosis and treatment experience of pelvic fracture complicated with anterior sacroiliac joint dislocation.Methods:The data of 19 patients with pelvic fracture and anterior sacroiliac joint dislocation admitted from June 2013 to September 2019 were retrospectively analyzed. There were 12 males and 7 females; aged 9-67 years, with an average of 28.0 years. There were 11 cases of traffic accident injury, 6 of falling injury and 2 of crashing injury caused by machine extrusion. According to the Tile classification, there were 8 cases of type C1, 2 of type C2, and 9 of type C3; 6 cases of modified mangled extremity severity score (MESS) were greater than or equal to 7, of which 2 cases underwent hip amputation, 4 cases underwent king-steelquis semipelvectomy. 13 cases underwent limb-salvage surgery, of which 3 cases underwent external fixation surgery, and the remaining 10 cases underwent open reduction and internal fixation with steel plate. The reduction quality was evaluated according to the Matta imaging criteria of pelvis, the pain was evaluated according to the visual analogue scale (VAS), and the pelvic function was evaluated according to the Majeed score.Results:The operation time was 2 to 4 hours, with an average of 3.2 hours; the intraoperative blood loss was 400 to 1 200 ml, with an average of 768.4 ml. The 18 surviving patients were followed up for 6 to 24 months, with an average of 11.6 months. One of the 6 amputation patients died one week after the operation due to septic shock and multiple organ failure. The overall survival rate of the patients was 94.7% (18/19), and the amputation rate was 31.6% (6/19). The pelvic fractures of the 18 surviving patients were all healed, and the fracture healing time was 3 to 6 months, with an average of 4.2 months. No delayed union or nonunion occurred. At the last follow-up of the 18 surviving patients, the VAS was 2.4±1.9 points (range, 0-6 points). Among them, there were no pain in 3 cases, mild pain in 9 cases and moderate pain in 6 cases. At the last follow-up, 13 patients undergoing limb salvage treatment evaluated the quality of reduction according to Matta imaging criteria. Among them, 6 cases were excellent, 4 were good, 2 were fair, and 1 was poor. The excellent and good rate was 76.9% (10/13). The Majeed functional score was 81.8±11.9 points (range, 53-95 points). Among them, 6 cases were excellent, 5 were good, 1 was fair, and 1 was poor. The excellent and good rate was 84.6% (11/13). In the limb salvage treatment group, 4 patients with lumbosacral nerve injury caused limited hip joint movement function, and unilateral lower limb sensation and movement were weakened. After neurolysis and nutritional support treatment, the patient's sensorimotor function was significantly improved.Conclusion:The mortality and disability rate of pelvic fractures combined with anterior sacroiliac joint dislocation is high. In order to save the patient's life, early diagnosis of limb injury and early surgical intervention should be performed. If necessary, hip disarticulation or king-steelquis should be selected.

Over the past three decades， China has made significant progress in the prevention and control of viral hepatitis， and the incidence rates of new-onset pediatric hepatitis B virus infections and acute viral hepatitis in the population have reduced to a relatively low level； however， there is still a heavy disease burden of chronic viral hepatitis in China， which severely affects the health status of the population. This study systematically summarizes the achievements of viral hepatitis prevention and control in China， analyzes existing problems and challenges， and proposes comprehensive prevention and control strategies and measures to eliminate viral hepatitis as a public health threat based on the national conditions of China， in order to provide a reference for related departments in China on how to achieve the action targets for eliminating viral hepatitis as a public health threat by 2030.