[Recommendations on the diagnosis and treatment of osteoarthritis in China].

Osteoarthritis (OA) is the most common form of arthritis and the leading cause of old age disability, affecting an estimated 302 million people worldwide. OA is seriously overlooked in the world. The awareness of OA and the popularization of standardized diagnosis and treatment are all lacking. Knees, hips, and hands are the most commonly affected joints in OA. Based on the experience of diagnosis and treatment, consensus and guidelines, we formulated this diagnosis and treatment standard in order to standardize the diagnosis and treatment of OA. We hope that our standard can reduce misdiagnosis and mistreatment and improve the prognosis of OA.

[Expression of annexin A1 in peripheral blood cells of Naïve rheumatoid arthritis patients and its influencing factors].

Objective: To explore the expression of Annexin a1 (AnxA1) mRNA in peripheral blood mononuclear cells (PBMC) from Naïve rheumatoid arthritis (RA) patients and healthy controls and to investigate its influencing factors. Methods: Thirty Naïve RA patients and 36 healthy controls were recruited from the First Affiliated Hospital of Harbin Medical University between May 2013 and January 2015.All RA patients were fulfilled the classification criteria of ACR in 1987.Clinical characteristics and laboratory indexes were collected.Real time polymerase chain reaction (RT-PCR) was used to measure AnxA1 and Foxp3 mRNA expression respectively.Serum TNF-α, IL-17 and IL-10 levels in each group was determined separately by enzyme linked immunosorbent assay (ELISA). Furthermore, correlations between the expression of AnxA1 mRNA and clinical characteristics, laboratory parameters, Foxp3 mRNA, serum IL-17, TNF-α, IL-10 level in Naïve RA patients were analyzed. Results: The expressions of AnxA1 mRNA in PBMC of Naïve RA patients were lower than that of healthy controls [0.12 (0.05-0.30) versus 1.66 (0.40-2.68), P<0.05]. The Foxp3 mRNA expressions in PBMC of Naïve RA patients were also lower than that of healthy controls [0.77 (0.39-1.89) versus 2.93 (2.65-3.49), P<0.05], while serum TNF-α, IL-17, and IL-10 levels of Naïve RA patients were higher than those of healthy controls (P<0.05). The expression of AnxA1 was positively associated with the expression of Foxp3 mRNA (r(s)=0.513, P<0.05) and negatively associated with the serum IL-17 level (r(s)=-0.381, P<0.05). But there were no correlations between expression of AnxA1 in PBMCs from Naïve RA patients with the TNF-α, IL-10 level, clinical characteristics (sex, age, disease duration and disease activity) and the laboratory indexes (RF, Anti-CCP antibody, ESR and CRP). The expression of Foxp3 mRNA and the serum IL-17 level was the independent influencing factor. Conclusions: The reduced expression of AnxA1 may associate with the reduced expression of Foxp3 and the increased IL-17 level in Naïve RA patients.It suggested that AnxA1 may play a key role in immune regulation and anti-inflammatory process in the pathogenesis of RA.

Application of a functional marker for the effect of cryoprotectant agents on gorgonian coral (Junceella juncea and J. fragilis) sperm sacs.

BACKGROUND: The establishment of coral sperm repositories which retain good post-rewarming viability and fertility play a vital role in species conservation. OBJECTIVE: This study aimed at obtaining baseline information regarding the effects of cryoprotectant agents (CPAs) on gorgonian coral (Junceella juncea and J. fragilis) sperm sacs. METHODS: The adenosine triphosphate assay was used to determine the energy level of the gorgonian sperm sacs as an indicator of sperm viability after exposure to cryoprotectants. RESULTS: The 'no observed effect concentrations' (NOECs) of methanol, dimethyl sulfoxide (DMSO), polypropylene glycol (PG), ethylene glycol (EG) and glycerol for J. juncea sperm sacs were 3 M, 3 M, 1 M, 2 M and 1 M respectively after 20 min exposure; whilst the NOECs for J. fragilis oocytes were 2 M, 3 M, 1 M, 2 M and 2 M, respectively. Methanol and DMSO had the least impact. PG was the most toxic CPA after 10 min exposure. ATP content of J. juncea and J. fragilis sperm sacs did not differ significantly from the control with incubation times of 10-20 min with 2 M EG. However, ATP content dropped significantly after exposing sperm sacs to 2 M EG for 40 min with average values of 2.34 +/- 0.12 and 1.97 +/- 0.48 microg/ml respectively. ATP content for J. juncea and J. fragilis sperm sacs was significantly decreased to 1.79 +/- 0.31 and 2.40 +/- 0.36 microg/ml after 20 min incubation in 2 M PG when compared to the control with 2.98 +/- 0.16 and 4.14 +/- 0.42 microg/ml respectively. Normalized ATP content for sperm sacs of two different gorgonian coral after incubation in methanol, DMSO, PG, EG and glycerol showed that J. juncea sperm sacs were slightly less tolerant to CPAs compared to J. fragilis sperm sacs. CONCLUSIONS: DMSO or methanol can be considered as efficient CPAs for gorgonian sperm sacs cryopreservation. The ATP luminescence assay provided sensitive and rapid quantification of mitochondrial activity in gorgonian coral sperm sacs. The study on the impact of CPA will contribute to the development of a cryopreservation protocol for coral sperm conservation.

[Depth of getting qi in clinical practice].

Based on the type of build being fat, medium and thin to determine the depth of getting Qi (DGQ) in acupoints, and based on the aforesaid three types of build to verify the difference of DGQ of neck, trunk, upper-limb and lower limb with T test. The result showed that the DGQ of fat man group was deeper, the DGQ of thin man group was more superficial (P < 0.01), the DGQ of neck was more superficial than that of trunk, upper and lower limb, and the standard deviation (SD) of neck DGQ was +/- 0.1 cm, that of trunk, upper and lower limbs was +/- 0.2cm. Following finding has been observed, the DGQ of patient with nervousness and allergic constitution showed more superficial, while the DGQ of cancer and apoplexy patients showed deeper than that of ordinary patients.

[Latin square vs twin crossover design for mouse blood glucose assay to estimate insulin potency].

According to parallel line analysis, a Latin square design was used for estimating insulin potency in mouse blood glucose assay. Four to six groups of 4 x 4 Latin squares were used to estimate 80%, 100% and 120% standard preparations and the recovery rates were 95-106%. Meanwhile, in comparison with twin crossover design, 13 batches of variant preparations including some beyond expiry dates were checked by Latin square design. The results showed that the potencies were about the same in two designs. The average fiducial limit rates were still less than 25% but 40% or so of animal numbers were saved. Therefore, Latin square design in a precise and accurate assay for the estimation of insulin.

[Multivariate twin crossover design parallel line assay for estimating potency of insulin].

Based on twin crossover design, a multivariate assay design was used in parallel line analysis to estimate the potency of insulin. This paper describes how to make the experimental design. After the analysis of variance according to assay, the potency and fiducial limits were calculated by the method (2.2). It has been applied by multivariate assay to estimate 80, 100 and 120% standard preparation and the recovery rate were between 94 and 108%. We also tested various batches and agent products. The average fiducial limits rates were less than 20%. In comparison with twin crossover analysis as usual, multivariate assay appeared to provide a more efficient utilization of experimental data, to increase weight of every animal for a more accurate estimation of potencies and to save on animal numbers 30-40%.

Site-specific mutagenesis of Drosophila alcohol dehydrogenase: evidence for involvement of tyrosine-152 and lysine-156 in catalysis.

Amino acid sequence comparisons reveal that tyrosine-152 and lysine-156 of Drosophila alcohol dehydrogenase (ADH) are conserved in homologous dehydrogenases, suggesting that these residues are important in catalysis. To test this hypothesis, we used site-directed mutagenesis to substitute tyrosine-152 with phenylalanine, histidine, or glutamic acid or to substitute lysine-156 with isoleucine. All of these mutants are catalytically inactive. Two mutants were active: A cysteine mutation of tyrosine-152 has 0.25% of wild-type ADH activity, while an arginine substitution of lysine-156 retains 2.2% of wild-type ADH activity. Kinetic analysis shows that the cysteine mutant increases Km(ethanol) 56-fold and Km(propan-2-ol) 100-fold, while Km(NAD) values are essentially unaltered. The arginine mutant also shows the significant enlargement of Km(ethanol), but not of Km(NAD). Furthermore, the cysteine mutant and arginine mutant have different substrate specificity and behave differently on competitive inhibition than wild-type ADH. These results suggest that both tyrosine-152 and lysine-156 have essential roles in catalysis by Drosophila ADH.