Objective:To investigate the expression of HERG1 and Kv3.4 in normal oral mucosa(NOM)and oral lichen planus (OLP).Methods:20 OLP and 1 6 NOM specimens were collected and immunohistochemically stained(IHC)by SP method for the detection of HERG1 and Kv3.4 protein expression.Results:HERG1 and Kv3.4 were negative or weak-positive in the sapmles of bas-al layer of NOMand non-erosive OLP,but positive in basal layer,spinous layer and granular layer of erosive OLP.The expression of HERG1 and Kv3.4 was higher in OLP than in NOM tissues (P <0.05);and higher in erosive OLP than in non-erosive OLP(P <0.05).In OLP HERG1 expression was positively related to Kv3.4(P <0.05).Conclusion:HERG1 and Kv3.4 may be related to the development of OLP.

Objective To evaluate the relationship between cold hyperalgesia and trafficking of transient receptor potential melastatin 8 (TRPM8) to cell membrane in the dorsal root ganglion (DRG) of rats with neuropathic pain (NP).Methods Ninety-six healthy male Sprague-Dawley rats,aged 10-12 weeks,weighing 250-280 g,were divided into sham operation group (S group,n=48) and NP group (n =48) using a random number table.NP was produced by chronic constriction injury to the sciatic nerve.The number of paw lifts on the cold plate and mechanical paw withdrawal threshold (MWT) were measured on 1 day before operation and 1,4,7,10 and 14 days after operation.Rats were sacrificed after behavioral testing,and ipsilateral DRGs of the lumbar segment (L46) were dissected tor detection of the expression of TRPM8 in total and membrane proteins by Western blot,and the ratio of TRPM8 expression in the membrane protein to that in the total protein (m/t ratio) was calculated.Results Compared with group S,the number of paw lifts on the cold plate was significantly increased,the MWT was decreased,the expression of TRPM8 in total and membrane proteins was up-regulated,and m/t ratio was increased on postoperative days 4,7,10 and 14 in group NP (P＜0.05 or 0.01).In group NP,the number of paw lifts on the cold plate was gradually increased with the prolongation of time after operation and reached the peak on postoperative day 10,maintaining at the peak until postoperative day 14;the MWT was gradually decreased and reached the lowest level on postoperative day 10,maintaining at the lowest level until postoperative day 14;the expression of TRPM8 in total and membrane proteins and m/t ratio were gradually increased with the prolongation of time after operation and reached the peak on postoperative day 10,maintaining at the peak level until postoperative day 14 (P＜0.01).Conclusion The mechanism underlying the development of cold hyperalgesia is related to enhanced trafficking of TRPM8 to cell membrane in DRGs of rats with NP.

OBJECTIVEThis study aims to test the accuracy and precision of iWitness photogrammetry for measuring the facial tissues of mannequin head.

METHODSUnder ideal circumstances, the 3D landmark coordinates were repeatedly obtained from a mannequin head using iWitness photogrammetric system with different parameters, to examine the precision of this system. The differences between the 3D data and their true distance values of mannequin head were computed.

RESULTSOperator error of 3D system in non-zoom and zoom status were 0.20 mm and 0.09 mm, and the difference was significant (P 0.05). Image captured error of 3D system was 0.283 mm, and there was no significant difference compared with the same group of images (P>0.05). Error of 3D systen with recalibration was 0.251 mm, and the difference was not statistically significant compared with image captured error (P>0.05). Good congruence was observed between means derived from the 3D photos and direct anthropometry, with difference ranging from -0.4 mm to +0.4 mm.

CONCLUSIONThis study provides further evidence of the high reliability of iWitness photogrammetry for several craniofacial measurements, including landmarks and inter-landmark distances. The evaluated system can be recommended for the evaluation and documentation of the facial surface.

Better governance of private hospitals is key to deepening the health reform.In view of the present development of private hospitals in Hubei province,this paper expounded the characteristics development of private hospitals and the governance of specific measures in the province and analyzed the defects found in the legal system and mechanism.The authors also raised such recommendations as perfecting the legal system and improving the mechanism of information disclosure and evaluation.

Objective To investigate the expression of the Kv3 .4 protein and Kv3 .4 mRNA in various stages of oral carcino‐genesis .Methods The expression of Kv3 .4 protein and Kv3 .4 mRNA were detected by immunohistochemistry (SP method) and RT‐PCR technique respectively in the oral carcinogenesis of SD rat which were induced by 4NQO .Results With the aggravation of the epithelial dysplasia ,Kv3 .4 protein and Kv3 .4 mRNA expression increased gradually .They were strongly positive in oral squa‐mous cell carcinoma .Conclusion The expression of Kv3 .4 protein and Kv3 .4 mRNA levels increased consistently with the aggra‐vation of the epithelial dysplasia .

Objective To prepare polyclactic-co-glycolic acid(PLGA) polymer ultrasound contrast agent loading sudan black dye (SB-PLGA) and study the enhancement effect of it on rabbit lymph nodes imaging and as a sentinel lymph node (SLN) biopsy in the feasibility and value of the indicator.Methods SB-PLGA ultrasound contrast agent was made by double emulsion and freeze-drying methods.Thigh subcutaneous rabbit VX2 tumor model of eight lymph nodes were set up with the foot pad subcutaneous injection of contrast agents to get contrast-enhanced ultrasound imaging of the popliteal lymph node and the second inguinal lymph node stations.Then the blue stained lymph nodes were resected by popliteal lymph node dissection and the second station lymph nodes were also observed after 30 mins respectively,lymph nodes were examined by frozen sections HE staining in parallel.Results SB-PLGA microbubbles had a tight size distribution.Ultrasound imaging can be significantly enhanced lymph node imaging,the second station lymph node imaging was not obvious.Popliteal lymph node dissection for lymph node staining,30minutes later,frozen sections HE staining were seen within the lymph node contrast agent present in the lymphatic sinus and a large entry of macrophages.Inguinal lymph node dissection showed the second station of lymph nodes no blue staining.Conclusions SB-PLGA ultrasound contrast agent is good for sentinel lymph node imaging and biopsy indicator.Contrast agent retention and accumulation in the lymph nodes by the uptake of macrophages may be associated with the mechanisms of lymph node enhanced imaging.

Objective To prepare a kind of superparamagnetic iron oxide (SPIO) and doxorubicin loaded multifunctional polymer microbubbles (MPMBs),and to explore its potential application as an ultrasound(US)/magnetic resonance (MR) imaging contrast agent in vitro.Methods The MPMBs and normal polymer microbubbles (PMBs) were made by double emulsion and freeze-drying methods.The physical property,drug encapsulation efficiency and the drug-loading efficiency of MPMBs were determined,and the release property and US/MR imaging enhancement of MPMBs were observed.Results The MPMBs had a regular shape and narrow size distribution.The drug encapsulation efficiency was (60.20±2.69) ％,and the drug-loading efficiency was (6.02 ± 0.27) ％.The in vitro release experiment showed that ultrasound can promote the release of doxorubicin in MPMBs.US imaging in vitro showed that the enhancement of MPMBs was better than PMBs,and MR imaging in vitro conformed that MPMBs could well enhance MR imaging.Conclusions The MPMBs is a multifunctional contrast agent with the treatment function as well as US/MR dual-mode imaging enhancement effect.

Objective To evaluate the effect of dexmedetomidine on necroptosis during liver injury in septic rats.Methods Eighteen SPF adult male Sprague-Dawley rats,weighing 200-220 g,were divided into 3 groups (n=6 each) using a random number table:sham operation group (group SH),sepsis group (group SEP) and dexmedetomidine group (group DEX).Sepsis was induced by cecal ligation and puncture in chloral hydrate-anesthetized rats in SEP and DEX groups.Dexmedetomidine 5 μg/kg was injected via the caudal vein at 1 h before operation in group DEX.Blood samples were collected from the caudal vein at 6 h after operation for determination of serum aspartate amino-transferase (AST) and alanine aminotransferase (ALT) concentrations.The rats were then sacrificed and livers were removed for determination of the level of reactive oxygen species (ROS) in liver tissues (using chemiluminescence assay) and expression of receptor-interacting protein 1 (RIP1),RIP3,mixed lineage kinase domain-like (MLKL),high-mobility group box 1 protein (HMGB1) and dynamin-related protein 1 (Drpl) in liver tissues (by Western blot).Results Compared with group SH,the serum AST and ALT concentrations were significantly increased,the expression of RIP1,RIP3,MLKL,HMGB1 and Drpl in liver tissues was up-regulated,and the level of ROS in liver tissues was increased in SEP and DEX groups (P＜0.05).Compared with group SEP,the serum AST and ALT concentrations were significantly decreased,the expression of RIP1,RIP3,MLKL,HMGB1 and Drp1 in liver tissues was down-regulated,and the level of ROS in liver tissues was decreased in group DEX (P＜0.05).Conclusion The mechanism by which dexmedetomidine attenuates liver injury may be related to inhibition of necroptosis in septic rats.

Objective To evaluate the role of D-serine in nerve cell apoptosis induced by multiple exposures to sevoflurane anesthesia in newborn mice and its relationship with glycogen synthase kinase-3 beta (GSK-3β).Methods Thirty healthy male C57B/L6 mice,aged 6 days,weighing 3.5-4.5 g,were divided into 3 groups (n =10 each) using a random number table:control group (group C),multiple exposures to sevoflurane anesthesia group (group S) and D-serine group (group D).On postnatal days 6,7 and 8,3％ sevoflurane in 30％ oxygen was inhaled for 2 h starting from 10:00 daily,and normal saline 0.1 ml and D-serine 500 mg/kg were intraperitoneally injected at 30 min before inhalation in S and D groups,respectively.In group C,30％ oxygen was inhaled for 2 h starting from 10:00 daily,and normal saline 0.1 ml was intraperitoneally injected at 30 min before inhalation.The animals were sacrificed after the end of oxygen or sevoflurane inhalation on postnatal day 8,and the brains were removed for determination of the expression of phosphorylated GSK-3β (pGSK-3β) and activated caspase-3 in brain tissues by Western blot.Results Compared with group C,the expression of pGSK-3β in brain tissues was significantly down-regulated,and the expression of activated caspase-3 in brain tissues was up-regulated in group S (P＜ 0.05),and no significant change was found in the parameters mentioned above in group D (P＞0.05).Compared with group S,the expression of pGSK-3β in brain tissues was significantly up-regulated,and the expression of activated caspase-3 in brain tissues was down-regulated in group D (P＜0.05).Conclusion D-serine is involved in the nerve cell apoptosis induced by multiple exposures to sevoflurane anesthesia through inhibiting the activation of GSK-3β in newborn mice.

Objective:A method that is based on microfluidic cell chip technology was developed for the first time to analyze CD14+monocyte myeloperoxidase (MPO) expression in myelomonocytic leukemia (M4) patients. CD14+monocyte MPO expression in M4 patients was preliminarily discussed. Methods:a. The chip was prepared by using polydimethylsiloxane as the host material and by secondary foam molding. b. A total of 48 clinically diagnosed M4 patients and 52 patients with normal myelogram were included as the test and control groups, respectively. c. A method based on the microfluidic cell chip approach was established to detect CD14+mono-cytes and to determine the positive rate and degree of MPO expression in the cells. d. The microfluidic cell chip technique was used to compare CD14+monocyte MPO expression in M4 patients with that in the control. Results:a. The designed microfluidic single cell analysis chip allowed the entry of granulocytes into the corresponding microfluidic channels. Thus, blood cells were separated. Numer-ous ghost corpuscles surrounded the separated white blood cells (WBCs). WBC morphology did not show obvious changes. b. The posi-tive rate of MPO expression and the activity of CD14+monocytes in the bone marrow of M4 patients were significantly higher than those in the bone marrow of the control (P<0.05). Conclusion:A method based on microfluidic single cell technology was developed for the first time to analyze the MPO expression in CD14+monocytes. CD14+monocyte MPO activity in M4 patients was significantly higher than in the control. CD14+monocyte MPO activity can be used as an auxiliary examination marker for clinical diagnosis.