BACKGROUND:Transient receptor potential protein channel is distributed in many kinds of tissue and cells in human body, and takes part in many physiologic functions and pathological changes of human body. However, up to now, transient receptor potential melastatin 8 (TRPM8) has not been reported in human cartilage.
OBJECTIVE:To study the expression of TRPM8, a subfamily member of transient receptor potential gene, in human knee joint cartilage.
METHODS:Fifteen samples were selected from patients after amputatioin due to severe trauma. Total protein was extracted from 300 mg cartilage tissue of every patient. To detect the distribution of TRPM8 in human
cartilage, cartilage sections were col ected in the same region. The expression and distribution of TRPM8 in human cartilage was detected by western blot analysis and immunofluorescence respectively.
RESULTS AND CONCLUSION:Western blot analysis showed that specific TRPM8 band was visible at molecular weight of 130 000, and as the age increased, TRPM8 expression in human cartilage was gradual y decreased. Immunofluorescence staining results showed that TRPM8 was evenly located in the cellmembrane of chondrocytes. TRPM8 is expressed in the human knee cartilage, specifical y in member of chondrocytes.

Objective To provide evidence for the application of nutritional intervention strategy by analyzing perioperative nutritional status of retroperitoneal liposarcoma patients.Methods 60retroperitoneal liposarcoma patients were divided into primary group (27 cases) and relapse group (33cases).These patients were put on nutritional risk evaluation using nutritional risk screening (2002)(NRS2002) within 24 hours of admission,and the serum albumin (ALB) value was recorded.Results Preoperative NRS2002 score showed that relapse group had a higher chance of cacotrophy.Serum albumin level from different time point showed that patients in relapse group were more prone to cacotrophy (respectively t =9.174,4.720,10.950,8.772,10.960,all P ＜ 0.05) and subject to slower recovery of gastrointestinal function [(86 ± 15) h vs.(61 ± 6) h] (t =7.903,P ＜ 0.05),prolonged antibiotic use [(15.5 ± 5.6) d vs.(7.3 ± 0.7) d],t =7.55,P ＜ 0.05),the average length of hospital stay was longer [(25.5±6.0) d vs.(14.1 ± 1.7)d] (t =9.571,P＜0.05),and higher inhospita cost [(13.2±5.6) × 104 yuan vs.(6.8 ± 0.9) × 104 yuan] (t =5.871,P ＜ 0.05).Though there was no difference between the two groups in incision complication rates (x2 =2.625,P ＞ 0.05)Conclusions Retroperitoneal liposarcoma patients has perioperative nutritional risk.Compared with the primary group,patients from relapse group have higher chance to got cacotrophy.NRS2002 scoring system is simple,it could be used in predicting nutritional risk for those patients.

Objective To evaluate the therapeutic effect of minimally invasive retractor reduction and fixation through sinus tarsi approach together with external application of Chinese medicine for displaced intra-articular calcaneal fractures. Methods From October of 2009 to June of 2013, 52 patients with displaced intra-articular calcaneal fractures (Sanders type Ⅱ, Ⅲ) were enrolled into the study. The patients received minimally invasive retractor reduction through sinus tarsi approach and fixation with small plate and cannulated screws, and after the operation were given external washing with Chinese herbal formula Shenxing Fang ( mainly composed of Herba Lycopodii, Herba Speranskiae Tuberculatae, Rhizoma Sparganii, Rhizoma Curcumae, Herba Asari, Radix Aconiti Preparata, Radix Aconiti Kusnezoffii Preparat). Follow-up was carried out for the evaluation of the therapeutic effect of displaced intra-articular calcaneal fractures ( Sanders type Ⅱ, Ⅲ). Results Forty cases (involving 44 feet) received the postoperative follow-up for 12-18 months (average being 14.2 months). The Maryland foot scores were 95 points for type Ⅱ displaced intra-articular calcaneal fractures, and 86 points for type Ⅲ fractures. Satisfactory results were achieved in the height, width and length of the calcaneus as well as the B?hler's and Gissane angle ( P<0.01). Conclusion Minimally invasive retracter reduction and fixation through sinus tarsi approach together with external application of Chinese medicine exert satisfying effect for the treatment of displaced intra-articular calcaneal fractures.

Objective To study the effect of lung cancer prevention in Baoji city, one of the cities with high lung cancer morbidity in China. Methods To investigate the mortality of lung cancer from 1992 to 1996 in Baoji and compare it with the data of Baoji in 1970s and the current data in whole China. Information of lung cancer pa- tients was collected from the Malignant Tunout Monitoring Station in Baoji. To investigate by sampling survey the status of smoking of Baoji population based on the principle of clustering random sampling. Information about air pollution in 1980s and 1990s was obtained from the Environmental Monitoring Station in Baoji. ResultsComparing with 1970s, the crude mortality rates of lung cancer in Baoji in 1992～ 1996 increased both in males and females,but the range was narrower than that in whole China. Ajusted mortality rates of lung cancer didn＇t change in females but decreased in males. However, either crude or adjusted mortality rate was lower than the average level of the whole country. The status of air pollution has been improved dramatically since the early 1980s in Baoji. So far, except to- tal suspended particles (TSP), NOx and SO2 levels have reached the Criteria of Air Quality of China. But the smoking rate was still higher. ConclusionThe prevention of lung cancer in Baoji is effective, but the role of measures needs further study. Baoji should not be considered as a city with high lung cancer morbidity in our country at present.

Objective To compare the effects of arthroscopic surgery plus one or more posterior small incisions and the sinus tarsi approach in the treatment of calcaneal fracture.Methods A retrospective analysis was conducted of the 85 patients with calcaneal fracture who had been treated from January 2017 to June 2017 at Department of Foot & Ankle Surgery,Guangzhou Orthopaedic Hospital.They were 43 men and 42 women,32 to 58 years of age (average,46.0 years).Arthroscopic surgery plus one or more posterior small incisions was performed in 40 of them while the sinus tarsi approach was used in the other 45 cases.The 2 groups were compared in terms of operation time,fracture healing time,incision complications and functions of the affected foot by the American Orthopedic Foot Ankle Society (AOFAS) ankle-hindfoot scores.Results There were no significant differences in the preoperative general data between the 2 groups,showing they were comparable (P ＞ 0.05).The average follow-up period for all the patients was 8 months (from 6 to 12 months).For the arthroscopic surgery group and sinus tarsi approach group,the fracture healing time was 8.6 ± 2.4 weeks and 8.9 ± 1.8 weeks,and the AOFAS ankle-hindfoot scores were 82.5 ± 5.6 and 85.1 ± 4.0,respectively,showing no significant differences between them (P ＞ O.05).The operation time in the arthroscopic surgery group (43.6 ±5.4 min) was significantly less than in the sinus tarsi approach group (56.5 ±6.4 minutes),and the rate of complications in the former[2.5％ (1/40)] significantly lower than in the latter[15.6％ (7/45)] (P ＜0.05).Conclusion Arthroscopic surgery plus one or more posterior small incisions may be a fine treatment for calcaneal fractures because postoperative incision complications can be reduced.

In order to further research into the EEG information of rats epileptic model, we applied different nonlinear dynamic methods. After having analyzed the EEG signal of rat falling sickness by means of approximate entropy and correlation dimension, we adopted a the new method, unstable periodic orbits, which was used to analyze complex activity of neurons system to look for the change regularity of change in the EEG signal in the whole course of rat's falling sickness. We found period 1 orbits and period 2 orbits to be statistically significant in the data of ictal time of epilepsy. At the same time, we found period 1 orbits to be statistically significant in the data of preictal time of epilepsy.
Animals ; Disease Models, Animal ; Electroencephalography ; Entropy ; Epilepsy ; physiopathology ; Nonlinear Dynamics ; Rats ; Signal Processing, Computer-Assisted

Objective:To select the differential prognostic lactic acid metabolism-related genes(LRGs)of the head and neck squamous cell carcinoma(HNSCC)to construct the LRGs prognostic model of HNSCC,and to clarify the potential mechanism.Methods:The HNSCC gene expression and clinical data were obtained from The Cancer Genome Atlas(TCGA)and Gene Expression Omnibus(GEO)Databases,the LRGs were identified through GeneCards Database,and R software was used to screen out the LRGs of HNSCC;univariate Cox regression analysis was used to identify prognosis-related genes;two different subtypes were identified based on the prognostis-related LRGs;Kaplan-Meier(K-M)curve analysis was used to compare the prognosis of the patients between two groups;CIBERSORT algorithm was used to perform the immuno-correlation analysis between two groups;multivariate Cox regression analysis and LASSO regression analysis were used to construct the prognostic model;receiver operating characteristic curve(ROC)and K-M survival curve were used to assess the relationship between LRGs and survival and prognosis of the HNSCC patients.The prognostic model was validated by GSE27020,GSE41613,and GSE65858 datasets.The experiment were grouped based on risk score,and immune-related analysis and tumor score analysis were performed.Results:The TCGA Database differential analysis results showed that 1 196 LRGs were identified from HNSCC samples;univariate Cox regression analysis selected 27 differentially expressed genes(DEGs)associated with the prognosis of the HNSCC patients.Two different LRGs subtypes(Group 1 and Group 2)were identified according to the prognosis-related genes.The K-M survival curves results showed that the overall survival(OS)of the patients in Group 2 was significantly higher than that in Group 1,and the immune cell expression amount of the patients in Group 2 was also higher than that in group 1.The multivariate Cox regression and LASSO regression analysis results screened out 9 LRGs,including hypoxanthine phosphoribosyltransferase 1(HPRT1),amyloid precursor protein(APP),glycogen phosphorylase L(PYGL),urokinase-type plasminogen activator(PLAU),cannabinoid receptor 2(CNR2),stanniocalcin 2(STC2),nucleotide binding oligomerization domain-like receptor protein 1(NLRP1),integrin-linked kinase(ILK),and forkhead box B1(FOXB1);the prognostic model was constructed.The K-M and ROC curve results indicated that the expression levels of above 9 genes were associated with the survival and prognosis of the HNSCC patients,providing good 1-year,2-year,and 3-year survival prediction effect,and the area under ROC curve(AUC)values were all greater than 0.650.Furthermore,the predictive ability of the prognosis model was validated in GSE27020,GSE41613,and GSE65858 datasets.The patients classified based on the risk scores had distinguishable immune statuses.Conclusion:The differentially expressed LRGs of HNSCC screened by bioinformatics methods are related to the survival and prognosis of the HNSCC patients;the prognostic model constructed by 9 LRGs can predict the survival status and treatment response of the HNSCC patients.

This study was aimed to investigate the effect of traditional Chinese medicine (TCM) compound "Ji-Sui-Kang (JSK)" on the differentiation of neural stem cells (NSCs).Drug serum of JSK was prepared.The primary NSCs were isolated and cultured.NSCs were identified.Fetal bovine serum (FBS) was used to induce differentiation of NSCs.Different doses of drug serum of JSK was added to intervene the differentiation.There were the high-,medium-,and low-dose group.The blank control group was also set.The β Ⅲ-tubulin and GFAP were used to detect the differentiation of NSCs.And then,it was compared with the control group without drug serum.Positive cells ofβ m-tubulin and GFAP in different visual fields were counted.And the percentage of positive cells in the total number of cells was calculated.Fluorescence microscope was used to conduct quantitative fluorescence experiments.The results showed that the cultured neurospheres were expressed of Nestin protein.The percentage ofβⅢ-tubulin positive cells in the high-,medium-and low-dose group of drug serum of JSK was significantly higher than that in the control group (P＜0.05).The percentage of GFAP positive cells in the high-,medium-and low-dose groups of drug serum of JSK was significantly lower than that in the control group (P＜0.05).And the high-dose group of drug serum of JSK was statistically significant compared with the middle-and low-dose group of drug serum of JSK (P＜0.01).It was concluded that TCM compound JSK promoted thedifferentiation of NSCs intoβⅢ-tubulin positive cells.It provided a basis for treatment of SCI with TCM.

β-glucosidase has important applications in food, pharmaceutics, biomass conversion and other fields, exploring β-glucosidase with strong adaptability and excellent properties thus has received extensive interest. In this study, a novel glucosidase from the GH1 family derived from Cuniculiplasma divulgatum was cloned, expressed, and characterized, aiming to find a better β-glucosidase. The amino acid sequences of GH1 family glucosidase derived from C. divulgatum were obtained from the NCBI database, and a recombinant plasmid pET-30a(+)-CdBglA was constructed. The recombinant protein was induced to express in Escherichia coli BL21(DE3). The enzymatic properties of the purified CdBglA were studied. The molecular weight of the recombinant CdBglA was 56.0 kDa. The optimum pH and temperature were 5.5 and 55 ℃, respectively. The enzyme showed good pH stability, 92.33% of the initial activity could be retained when treated under pH 5.5-11.0 for 1 h. When pNPG was used as a substrate, the kinetic parameters K_m, V_max and K_cat/K_m were 0.81 mmol, 291.99 μmol/(mg·min), and 387.50 s^-1 mmol^-1, respectively. 90.33% of the initial enzyme activity could be retained when CdBglA was placed with various heavy metal ions at a final concentration of 5 mmol/L. The enzyme activity was increased by 28.67% under 15% ethanol solution, remained unchanged under 20% ethanol, and 43.68% of the enzyme activity could still be retained under 30% ethanol. The enzyme has an obvious activation effect at 0-1.5 mol/L NaCl and can tolerate 0.8 mol/L glucose. In conclusion, CdBglA is an acidic and mesophilic enzyme with broad pH stability and strong tolerance to most metal ions, organic solvents, NaCl and glucose. These characteristics may facilitate future theoretical research and industrial production.
beta-Glucosidase ; Sodium Chloride ; Temperature ; Glucose ; Ethanol/chemistry* ; Ions ; Hydrogen-Ion Concentration ; Enzyme Stability ; Substrate Specificity

The incidence of each of the rare disease is very low. The complexity and diagnosis difficulty of the rare disease lead to the difficulties in the clinical research and development (R&D) of drugs for rare diseases. There is an urgent clinical need for the drug development of rare diseases in China. Encouraging R&D of new drugs, particularly the innovative drugs with China's own independent intellectural property is the basis for solving the predicament in drug shortage in China.. In order to further improve the efficiency of clinical R&D of drugs for rare diseases, the National Medical Products Administration (NMPA), Center for Drug Evaluation (CDE) issued Technical Guidance for Clinical Research and Development of Drugs for Rare Diseases. This is the first guidance for rare diseases in China that is drafted from the standpoint of the clinical technology research and development.The guidance is the scientifitc thinking and framework for the drug developing enterprises to research and develop drugs for rare disease efficiently and appropriately by following drug developing protocols and relating to the special features of rare disease.This paper presents the concepts and rationale in the guidance for the appraisal of rare disease drug research and development.