AIM:To clarify the impact of heart shock factor 4b (Hsf4b) K65R mutation on the regulation of downstream protein expression .METHODS:Non-functional Lys mutant plasmid pWZL-blast-HA-Hsf4b/K65R was genera-ted by replacing single , homologous amino acids using KOD-Plus-Mutagenesis-Kit.Mouse lens epithelial mLEC stable cell lines expressing Hsf4b or Hsf4b/K65R were constructed by lentivirus infection .The expression of Hsf4b in the mutant and the wildtype mLEC cells was confirmed by Western blotting .The expression of Hsf4b downstream proteins such as heat shock protein ( Hsp)70, Hsp90, Hsp27 and CryAB was examined by Western blotting and real-time PCR.RESULTS:The results of PCR and DNA sequencing confirmed the successful construction of mLEC Hsf 4b/K65R mutant.The K65R mutation didn’t influence Hsf4b expression in the mLEC cells.After K65R mutation in Hsf4b, the expression levels of Hsp27 and CryAB were down-regulated and the expression of Hsp 70i and Hsp90a upregulated.CONCLUSION: pWZL-blast-HA-Hsf4b/K65R can be used to construct a stable cell line by infecting with lentivirus .Hsf4b/K65R mutation influ-ences the regulation of downstream heat shock proteins .

BACKGROUND:Three-dimensional finite element simulation has been widely used in biomechanics. However, there is little research in knee joint replacement. The study of femoral prosthesis is less.
OBJECTIVE:To analyze the optimal positioning parameters of knee replacement and femoral prosthesis implantation, and to verify the efficacy of Gemini-PS knee prosthesis in total knee arthroplasty.
METHODS: (1) Knee finite element model in total knee replacement was constructed. Orthogonal experiment was performed in femoral osteotomy positioning parameters. The shift amount A, external rotation degree B, and valgus degree C of femoral prosthesis were selected as the relevant factors for orthogonal experiment. Three values were taken for each parameter to establish orthogonal tables. Nine experimental combined knee replacement finite element models were created and analyzed with finite element. Analysis of variance and range analysis were used by optimization. (2) Totally 42 patients (47 knees) with knee osteoarthritis received total knee arthroplasty with Gemini-PS knee joint prosthesis. Knee joint function was evaluated after replacement with Hospital for Special Surgery knee score and American Knee Society knee score. Knee pain was assessed with Visual Analogue Scale score.
RESULTS AND CONCLUSION:(1) Minimal polyethylene substrate surface compressive stress peak was translation 0 mm, external rotation of 3°, valgus 6°; the peak stress was 15.9 MPa. Among the influential factors, the effects of internal and external translation were larger than that of external rotation angle, and greater than that of valgus angle. The optimal positioning parameter combination was pan 0 mm, external rotation 3°, valgus 6° by range analysis and variance analysis. Orthogonal experiment was valid by simulation. (2) 42 patients were folowed up for 12-36 months. One patient affected subcutaneous fat liquefaction of the lower segment of the incision. Hospital for Special Surgery knee score and American Knee Society knee score were significantly higher after implantation (P < 0.05). Visual Analogue Scale score was significantly lower after replacement (P < 0.05). X-ray films showed that osteolysis, dislocation or loosening was not found. The recovery of knee joint was good after operation. (3) These findings

Objective:To analyze the clinical characteristics of patients with femoral neck fracture in Tianjin Hospital.Methods:Data of femoral neck fracture patients discharged from Tianjin Hospital from January 1, 2016 to December 31, 2017 were retrospectively analyzed. The gender, age, fracture type, injury mechanism and treatment method were collected.Results:Total of 2,150 eligible patients including 736 males and 1414 females were selected. There were significant differences in the gender distribution in different age groups ( χ2=91.63, P<0.001). Among young patients, males were more than females, while among middle-aged and elderly patients, females were more than males. The main type of fracture was displaced type (75.56%, 1,646/2,150), the displaced rate was the lowest in young patients (55.17%, 64/116), while it was the highest in old patients (81.91%, 1,159/1,415). The main injury mechanism of the three age groups were falls [young 61.21% (71/116), middle 80.29% (497/619), eldly 91.24% (1,291/1,415)]. There was a significant difference in the treatment methods between different age groups ( χ2=1,057.11, P<0.001). The main treatment method for young and middle-aged patients was internal fixation, no matter whether the fracture was displaced or not. The proportion of undisplaced patients with internal fixation (86.22%, 169/196) was higher than that of displaced patients with internal fixation (62.88%, 266/423) in middle-aged group ( χ2=34.93, P<0.001). In the elderly, more patients used internal fixation in undisplaced group, while more patients used hemiarthroplasty in displaced group. The age of old femoral neck fracture (median age was 74) was older than that of fresh fracture (median age was 70) ( Z=-2.777, P=0.005). And the displaced rate of patients with old femoral neck fracture (92.41%, 73/79) was higher than that of the fresh ones (75.95%, 1,573/2,071)( χ2=11.48, P=0.001). The patients with old femoral neck fracture usually adopt total hip replacement, while the proportion of three kinds of operation (internal fixation, hemiarthroplasty, total hip replacement) were similar among the fresh ones, and the internal fixation was the most. Conclusion:Femoral neck fracture is the most common in elderly women. The patients with displaced fracture are more than that with undisplaced fracture. Falling is the main injury mechanism in both young and old people. Internal fixation is commonly used in young and middle-aged patients, while arthroplasty is often used in elderly patients.

Objective To screen the human proteins interacting with human cytomegalovirus（HCMV）UL130 from human fetus brain cDNA library by GAL4 two-hybrid system 3 technique and analyze the corresponding coding sequences.Methods The ＂bait plasmid＂（named as pGBKT7-UL130）was constructed.By using HCMV UL130 as the bait,a human fetus brain cDNA library was screened and the proteins interacting with UL130 protein were searched.The positive clones were sequenced and analyzed by bioinformatic methods.Results Nine clones interacting with HCMV UL130 were identified,two of them were synaptosome-associated protein（SNAP）.Conclusion Some proteins interacting with HCMV UL130 in human fetus brain cDNA library were successfully screened.SNAP might play an important role in HCMV infection pathogenesis.

Objective Using yeast two-hybrid system to screen the proteins which can interact with the human cytomegalovirus (HCMV) UL128 which have two difference transcription structure from human fetus brain cDNA library, and compare the difference with structure and function of interacting proteins. Methods Two fragments of UL128 were amplified by 3'RACE and 5'RACE technology, the length are 519 bp and 642 bp, respectively. The "bait plasmid" (named as pGBKT7-UL128-519 bp and pGBKT7-UL128-642 bp) was constructed successfully. Using pGBKT7-UL128-519 bp and pGBKT7-UL128-642 bp as a bait, a human fetus brain cDNA was screened and the proteins interacting with UL128-519 bp and UL128-642 bp encoded protein were searched, and the positive clones were sequenced and analyzed by bioinformatic methods. Results EFEMP2 interacting with HCMV UL128-519 bp were identified, THY-1 interacting with HCMV UL128-642 bp were identified. Conclusion EFEMP2 and THY-1 proteins interacting with HCMV UL128-519 bp and UL128-642 bp in human fetus brain cDNA library were successfully screened, but same proteins weren't found from the proteins interacting with UL128-519 bp and UL128-642 bp protein, UL128-519 bp and UL128-642 bp protein may be play an different effect in the process of infect by HCMV.

Background With the development of phacoemulsification surgery,the minimization of the size of the incision in order to reduce the damaging of eye tissue and postoperative complications becomes a focus.Objective This study aimed to assess the clinical efficacy of coaxial phacoemulsification type cataract surgery through a 1.8 mm microincision with foldable intraocular lens (IOL) implantation in eyes with age-related cataract.Methods Informed consent was obtained from each patient at the beginning of this study.A serial case observation study was designed.Thirty-two eyes of 32 patients with age-related cataract were enrolled in the Inner Mongolia Autonomous Region People' s Hospital from April 2012 to May 2012,including 8 eyes of grade Ⅱ hard nucleus cataract,16 eyes of grade Ⅲ hard nucleus cataract and 8 eyes of grade Ⅳ hard nucleus cataract.Ocular axial length was measured by an A-mode ultrasonic apparatus,and IOL diopter was calculated using the SRK-Ⅱ formula.Under ocular surface anesthesia,a 1.8 mm clear corneal tunnel incision was made at the 10-11 o' clock position,and then an auxiliary incision was made at the 2 o' clock position.The opaque lens was extracted by routine phacoemulsification.An Akreos MI60 IOL was implanted through the 1.8 mm incision.The time and level of ultrasonic power required for phacoemulsification,postoperative visual acuity and incidence of postoperative complication were assessed.Postoperative examinations were scheduled at 1 day,1 week and 1 month after the surgery.Results All the operations proceeded smoothly.The mean phaco-time was (7.0±3.6) seconds and the mean ultrasonic power level was (15.3 ±6.1)％ among the different grades of cataract groups.The number of eyes presenting an uncorrected distance visual acuity of ≥ 0.5 was 18 (56.25％),28 (87.50％) and 30 (93.75％) 1 day,1 week and 1 month after surgery,respectively.The number of eyes with a best corrected distance visual acuity of ≥ 0.5 and 0.8 were 31(96.88％) and 26 (81.25％),respectively,1 month after surgery.Negligible changes were detected in the anterior chamber depths,and no thermal damage was found at the incision during the operation.Conclusions Coaxial phacoemulsification and IOL implantation through a 1.8 mm microincision is safe and effective.Thorough planning and precise execution are necessary.

Objective:To investigate effects of hydrogen sulfide (H2S) on inducible nitric oxide synthase (iNOS) in kidneys of Type 1 diabetic rats.Methods:Thirty-two male SD rats were randomly divided into four groups:A normal control (NC) group,a diabetes mellitus (DM) group,a NaHS (NaHS+DM) group,and a NaHS control (NaHS) group (n=8 per group).Type 1 diabetes was induced by a single intraperitoneal injection of streptozotocin (55 mg/kg).After successful establishment of models,the rats in NaHS+DM and NaHS groups were injected with NaHS solution (56 μmol/kg) intraperitoneally.Eight weeks later,the activities of total nitric oxide synthase (T-NOS) and iNOS,as well as the level of nitric oxide (NO) were detected in serum and renal tissues,respectively,The activity of glutathione peroxidase (GSH-Px) was determined in renal tissues,The ultrastructures of renal tissues were observed by transmission electron microscope,The protein expression ofiNOS in renal tissues was detected by Western blot.Results:Compared with the NC group,there was no significant difference in the various indexes in the NaHS group (P＞0.05).However,in the DM group,the activities of T-NOS and iNOS,and the level of NO were all increased significantly in serum and renal tissues,while the activity of GSH-Px was decreased in renal tissues.Under the electronic microscope,the thickening of the glomerular capillary basement membrane,the proliferation of mesangial matrix,and the foot fusion were observed.The protein expression ofiNOS was increased obviously in renal tissues in the DM group (P＜0.01).Compared with the DM group,the activities of T-NOS and iNOS and the level of NO were all decreased in serum and renal tissues,while the activity of GSH-Px was increased in renal tissues in the NaHS+DM group (P＜0.01).The renal ultrastructural damages were ameliorated obviously.The protein expression ofiNOS was decreased significantly (P＜0.01).Conclusion:H2S exerts a protective effect on kidney injury in type 1 diabetic rats.The mechanism might be related to inhibition of iNOS activity and protein expression,in turn leading to reduction of NO content in renal tissues.

Objective To investigate and analyze the military operation ability and influence factors of soldiers alongthe border in an extremely cold environment and to provide a scientific basis for effectively improving the combat effective -ness of the army stationed the cold regions.Methods According to relevant standards, recruits(the length of service <1month) and old soldiers(the length of service >1 year) were randomly selected to investigate their military operationfactors, physical condition, sleep condition, mental health and cognitive ability.Results Cold was the main factor whichaffected military operation capability in cold regions.According to physical standards,border troops were in poor physicalcondition.Results of SCL-90 showed that the total score, the number of positive items , the score of each factor of the 130recruits were significantly lower than those of the Chinese army model and 46 veterans (P <0.05).There was nosignificant difference in these aspects between the 46 veterans and Chinese army model.Recruits were in better sleepcondition and had better instantaneous memory(P <0.05).Birthplace had no effect on these factors .Recruits from Centraland South China were in poorer sleep condition than those from cold regions (P <0.05).Results of SCL-90 showed that thetotal score,the factors of somatization, coercion, depression and anxiety of communication veterans were significantly higherthan those of the Chinese army model (P <0.05).The sleep condition of communication veterans was poorer than that ofmotorized infantry and patrol veterans (P <0.05).Conclusion Cold is the main factorthat impacts the ability of militaryoperations in cold regions.The physical work capacity of border guards in cold regions was significantly below the militarystandard, so the level of military training should be strengthened .Research on cold-protection equipment for special tasks should be strengthened.

Objective: To discuss the surgical treatment of carcinoma of the pancreas with superior mesenteric artery (SMA) invasion. Methods: Vascular resection and reconstruction of the portal vein and hepatic artery were performed in 2 cases. Results: The portal vein (PV), superior mesenteric vein (SMV) and superior mesenteric artery (SMA) were involved. During surgery, the invaded superior mesenteric vessels were localized. Case 1 underwent pancreaticoduodenectomy with vascular resection, SMA partial resection and reconstruction. Case 2 was submitted to total pancreatic resection with simultaneous vascular resection of spleen artery and SMA reconstruction. No perioperative mortality occurred. Conclusion: Regional pancreaticoduodenectomy with superior mesenteric artery resection is a safe and effective surgical treatment for pancreatic carcinoma.

Objective To investigate the effect of plateau training on military operation ability ,evaluate the real situa-tion of combat forces on the plateau , and to provide data for effectively improving the combat effectiveness of the army stationed on the plateau .Methods According to the standards , the levels of high altitude acclimatization , physical work capacity (VO2max, PWC170) and mental work capacity (digit span, digit symbol, pursuit aiming and visual reaction time) were measured to assess the military operation ability of plateau training troops , including troops that entered the plateau from the plain(plateau-entering troops), troops stationed on the plateau (plateau troops) and troops stationed in the plain ( plain troops ) .Results The troops on the plateau all reached basic acclimatization .The aerobic capacities of these three types of troops were of the standard medium level , and the maximum oxygen uptake ( VO2max ) of the plain troops was greater than that of that of plateau-entering troops(P<0.01), and the VO2max of the plateau troops was greater than that of plateau-entering troops(P<0.01).Compared with the plain troops, the VO2 max and PWC170 of the plateau-entering troops decreased by 25.7%and 27.7%respectively.There was no significant difference in the digit span , digit symbol, pursuit aiming between these 3 kinds of troops , but the visual response of the plateau-entering troops was prolonged ( P<0.01), while the pursuit aiming of the plateau troops was lower than that of the plateau-entering troops(P <0.05). Conclusion Acclimatization-promoting measures and plateau training can contribute to high altitude acclimatization formation, but cannot overcome the objective adverse effects of the plateau on physical fitness .Two months of plateau training fails to improve physical fitness.Therefore, special attention should be paid to the research on scientific and effective measures to improve physical fitness on the plateau .