Caloric restriction (CR) is now a popular lifestyle choice due to its ability in experimental animals to improve lifespan, reduce body weight, and lessen oxidative stress. However, more and more emerging evidence suggests this treatment requires careful consideration because of its detrimental effects on the skeletal system. Experimental and clinical studies show that CR can suppress bone growth and raise the risk of fracture, but the specific mechanisms are poorly understood. Reduced mechanical loading has long been thought to be the primary cause of weight loss-induced bone loss from calorie restriction. Despite fat loss in peripheral depots with calorie restriction, bone marrow adipose tissue (BMAT) increases, and this may play a significant role in this pathological process. Here, we update recent advances in our understanding of the effects of CR on the skeleton, the possible pathogenic role of BMAT in CR-induced bone loss, and some strategies to mitigate any potential side effects on the skeletal system.

OBJECTIVE: To explore the genotype mutation characteristics of patients with glucose-6-phosphate dehydrogenase(G6PD) deficiency in Wuhan. METHODS: A total of 1 321 neonates with positive screening and outpatients were received G6PD mutation detection, 12 kinds of common G6PD mutation in Chinese people was detected by using multicolor melting curve analysis (MMCA) method, for those with negative results, the enzyme activity and clinical information were analyzed, sequencing was recommended after informed consent when it is necessary. RESULTS: Among 1321 patients, a total of 768 mutations were detected out, with a detection rate of 58.1%. A total of 18 types of G6PD genotypes were identified, including c.1388G>A, c.1376G>T, c.95G>A, c.1024C>T, c.871G>A, c.392G>T, c.487G>A, c.1360C>T, c.1004C>A, c.517T>C, c.592C>T, c.94C>G, c.152C>T, c.320A>G, c.1028A>G, c.1316G>A, c.1327G>C and c.1376G>C, including 683 male hemizygotes, 3 female homozygotes, 80 female heterozygotes and 2 female compound heterozygous. CONCLUSION A total of 18 types of G6PD mutations are identified in the reaserch, and c.94C>G, c.1028A>G and c.1327G>C are first reported in Chinese population. The most common G6PD mutation types in Wuhan are c.1388G>A, c.1376G>T, c.95G>A.
Asians/genetics* ; Female ; Genotype ; Glucosephosphate Dehydrogenase/genetics* ; Glucosephosphate Dehydrogenase Deficiency/genetics* ; Heterozygote ; Humans ; Infant, Newborn ; Male ; Mutation

OBJECTIVE：To st udy preventive effect and mec hanism of ginsenoside Rg 1 on focal cerebral ischemia-reperfusion injury（CIRI）model rats. METHODS ：Totally 78 SD rats were randomly divided into sham operation group ，model group ， butylphthalide control group （positive control ，10 mL/kg），ginsenoside Rg 1 low-dose，medium-dose and high-dose groups （10， 20，40 mg/kg），with 13 rats in each group. Administration groups were give relevant medicine intraperitoneally ，sham operation group and model group were given constant volume of normal saline intraperitoneally ，once a day ，for consecutive 7 d. After medication，except for the sham operation group ，focal CIRI model was induced by middle cerebral artery occlusion （MCAO） method in other groups. After modeling ，neurological deficit scoring was performed according to the modified neurological difict scoring standard ； TTC staining was used to detected the percentage of cerebral infarction of rats ；the cerebral water content was measured by dry/wet weight method ；serum contents of IL- 1β and IL-6 were detected by ELISA ；the protein expressions of p-p 38 MAPK and p-NF-κB p65 in cerebral tissue were determined by immunohistochemistry and Western blotting assay. RESULTS ： Compared with sham operation ，neurological deficits score ，percentage of cerebral infarction and cerebral water content ，serum contents of IL- 1β and IL-6，positive expression numbers of cells and protein expressions of p-p 38 MAPK and p-NF-κB p65 in cerebral tissue were increased significantly in model group （P＜0.05 or P＜0.01）. Compared with model group ，above index levels of administration groups were all decreased significantly （P＜0.05 or P＜0.01），and the effect of ginsenoside Rg 1 had a dose-dependent trend ；there was no significant difference of all above indexes between ginsenoside Rg 1 middle-dose，high-dose groups and butylphthalide control group （P＞0.05）. CONCLUSIONS ：Ginsenoside Rg 1 has a certain preventive effect on focal CIRI model rats ，the mechanism of which may be associated with down-regulating the protein expression of p-p 38 MAPK and p-NF-κB p65，inhibiting the release of inflammatory factors such as IL- 1β and IL-6.

OBJECTIVE: To investigate the effect of Chinese herbal formula Ermiao Powder (, EMP) on the expression of cholinergic anti-inflammatory pathway in rats with rheumatoid arthritis (RA). METHODS: Seventy-two rats were randomly divided into 6 groups according to body weight, including normal control group, collageninduced arthritis (CIA) group, three doses EMP groups, and methotrexate (MTX) group (n=12 per group). All of the rats except for those in the normal control group were given multipoint subcutaneous injection of bovine type II collagen to establish a CIA model. Three EMP groups received a high- (4.5 g/kg), medium- (3.0 g/kg), and low- (1.5 g/kg) doses of EMP by intragavage, respectively. MTX group was injected intraperitoneally MTX at 0.9 mg/kg once a week as the positive control. The administration was 3 consecutive weeks. Joint swelling, arthritis index, and body weight changes in different experimental groups of rats were tested. The joint damage was evaluated by masson staining. Quantitative real-time polymerase chain reaction, Western blot, and immunohistochemistry (IHC) were performed to evaluate the expression of CHRNA7, encoding α7 nicotinic acetylcholine receptor in the cholinergic anti-inflammatory pathway, in different tissues and their localization in the spleen and joints. RESULTS: CHRNA7 expression levels were significantly higher in the joints and spleens of CIA group than those in normal control group (both P<0.05). Moreover, the CHRNA7 mRNA and protein levels in the spleen and joints of MTX and three doses of EMP groups were significantly lower than CIA group (all P<0.05). Compared with the MTX group, treatment with low-dose EMP resulted in significant reduction of CHRNA7 mRNA and protein expression levels (P<0.05 or P<0.01). IHC showed positive signals of CHRNA7 in the white pulp and red pulp of the spleens of rats; CHRNA7 was expressed on fibroblast-like synoviocytes, macrophages, and endothelial cells in the joints of rats, and the expression in the joints of low-dose EMP group was significantly lower than that in the CIA group (P<0.01). CONCLUSIONS Cholinergic anti-inflammatory pathway was involved in the generation of the inflammatory reaction in CIA rats, and EMP exerted therapeutic effect on RA through cholinergic anti-inflammatory pathway.

OBJECTIVE：To comp are cytotoxicity and anti-inflammatory effects of raw Aconitium kusnezoffii and A. kusnezoffii processed with Terminalia chebula . METHODS ：Using H 9c2 cardiomyocytes isolated from rat as subjects ，CCK-8 assay was used to detect the effects of 0.5，1，2，4，6，8，10 mg/mL raw A. kusnezoffii and A. kusnezoffii processed with T. chebula on cell inhibition rate after cultured for 4，8，12，24 h. Hoechst 33258 staining was used to observe the effects on cell morphology characteristics after treated with 2，4，6 mg/mL raw A. kusnezoffii and A. kusnezoffii processed with T. chebula . Using macrophages RAW264.7 cells as subjects ，CCK-8 assay was used to detect the effects of 0.05，0.1，0.25，0.5，0.75，1，1.5，2 mg/mL raw A. kusnezoffii and A. kusnezoffii processed with T. chebula on cell survival rate after cultured for 24 h. ELISA assay was used to detect the effects of 0.05，0.1，0.25，0.5 mg/mL raw A. kusnezoffii and A. kusnezoffii processed with T. chebula on the release of NO ， TNF-α and IL-6 in RAW 264.7 inflammation cells induced by LPS. RESULTS ：When the mass concentration was 0.5，1 mg/mL， neither raw A. kusnezoffii and A. kusnezoffii processed with T. chebula had no inhibitory effect on H 9c2 cells. When the mass concentration was 2 mg/mL，the inhibitory effects of A. kusnezoffii processed with T. chebula on H 9c2 cells was higher than that of raw A. kusnezoffii （P＜0.05 or P＜0.01）；fluorescence intensity of cells treated for 24 h was stronger than that of raw A. kusnezoffii，but its nucleus was intact. When the mass concentration was 4-10 mg/mL，the inhibitory rate of A. kusnezoffii processed with T. chebula on H 9c2 cells at different time points （except for 24 h culture of 8，10 mg/mL）was significantly lower than raw A. kusnezoffii （P＜0.05 or P＜0.01）. The characteristics of cell morphology also showed that the fluorescence intensity of raw A. kusnezoffii group at 4，6 mg/mL was stronger than that of A. kusnezoffii processed with T. chebula group，and the cell nucleus fragmentation was more serious in the raw A. kusnezoffii group. 0.05-0.5 mg/mL raw A. kusnezoffii and A. kusnezoffii processed with T. chebula had no toxicity to RAW264.7 cells. 0.25，0.5 mg/mL raw A. kusnezoffii and 0.1，0.25，0.5 mg/mL A. kusnezoffii processed with T. chebula showed significant inhibitory effect on the release of NO ，0.05，0.1，0.25，0.5 mg/mL raw A. kusnezoffii and A. kusnezoffii processed with T. chebula showed significant inhibitory effect on the release of TNF-α and IL-6 in RAW 264.7 cell（P＜0.05 or P＜ 0.01）. The inhibitory effects of A. kusnezoffii processed with T. chebula at the same concentration on the release of NO was better than that of raw A. kusnezoffii ，but poorer than raw A. kusnezoffii in the inhibitory effects on the release of TNF-α and IL-6. CONCLUSIONS：The toxicity of A. kusnezoffii can be reduced after processed with T. chebula ，especially the toxicity of it in medium or high concentration and short-term use is lower than that of raw A. kusnezoffii . At the same time ，the anti-inflammatory effect of A. kusnezoffii processed with T. chebula is comparable to that of raw A. kusnezoffii at the same concentration.

OBJECTIVE：To compare the efficacy and safety of vancomycin given by continuous infusion vs. intermittent infusion，and to provide evidence-based reference for clinical drug use. METHODS ：Retrieved from PubMed ，the Cochrane Library，Embase，Wanfang database ，CNKI and VIP databases ，ranomized controlled trials （RCT）and cohort studies about vancomycin given by continuous infusion （trial group ）vs. intermittent infusion （control group ）were collected during the inception to Apr. 2020. After literature screening and data extraction ，the qualities of RCTs were evaluated by using bias risk evaluation tool recommended by Cochrane system evaluator manual 6.0. The qualities of cohort studies were evaluated by NOS ；Rev Man 5.3 software was used to perform Meta-analysis and publication bias analysis. RESULTS ：A total of 20 studies were included （3 RCTs and 17 cohort studies ），involving 2 380 patients in total. Results of Meta-analysis showed that ，target concentration attainment rate [RR ＝1.24，95％CI（1.12，1.38），P＜0.000 1] and attainment rate of target clinical efficacy [RR ＝1.20，95％CI（1.04，1.38）， P＝0.01] of trial group was significantly higher than those of control group. The incidence of nephrotoxicity [RR ＝0.56，95％CI （0.45，0.70），P＜0.000 01] was significantly lower than control group. There was no statistical significance in the therapeutic efficiency [RR ＝1.02，95％CI（0.95，1.10），P＝0.53]，drug treatment duration [MD ＝－0.50，95％CI（－1.40，0.39），P＝0.27] or mortality [RR ＝1.03，95％CI（0.78，1.35），P＝0.83] between 2 groups. The results of publication bias showed that the probability of publication bias was high when the incidence of nephrotoxicity was used as the index. CONCLUSIONS ：Vancomycin continuous infusion can improve the attainment rate of target concentration and target clinical efficacy ，reduce the incidence of nephrotoxicity ， but can not improve the treatment efficiency. Due to the inconsistent results of publication bias analysis ，the above conclusion needs to be interpreted carefully.

OBJECTIVE： To compare low-polarity volatile constituents in supercritical CO2 extract from the roots and stem of Ilex asprella and its effects on the proliferation of IEC-6 in vitro， and to provide reference for making full use of wild resources of I. asprella and expanding its medicinal parts. METHODS： The low-polarity volatile constituents were extracted from the root and stem of I. asprella with supercritical fluid CO2 extraction（SFE-CO2）. The chemical constituents were analyzed by GC-MS. IEC-6 cells were treated with different concentrations of supercritical CO2 extracts （0， 1， 5， 10， 20， 40， 60， 80， 100 μg/mL） from roots or stems of I. asprella. MTT assay was used to detect the relative viability， and cell proliferation curve was drawn and EC50 of each extract were calculated. RESULTS： Sixty-two and forty-six low-polarity volatile constituents were identified from supercritical CO2 extract in the roots and stem of I. asprella with GC-MS； there were 24 common constituents totally， mainly including pelargonic acid（14.18％ and 6.14％），octanoic acid（10.59％ and 4.35％），hexanoic acid（8.63％ and 10.86％），paeonol（7.79％ and 6.00％），2-methyl-3-phenyl-propanal（6.3％ and 0.58％），acetic acid（1.72％ and 33.77％） in root and stem， respectively. The results of cell culture in vitro showed that when the concentration of supercritical CO2 extract from the roots and stems of I. asprella was lower （≤60 μg/mL）， it could significantly promote the proliferation of IEC-6 cells and their EC50 were 16.35， 20.20 μg/mL， respectively； when the concentration of the extract was higher （≥80 μg/mL）， it showed cytotoxicity and inhibited the proliferation of IEC-6 cells. CONCLUSIONS： There are similar species of volatile constituents in roots and stems of I. asprella and similar in vitro bioactivity of the supercritical CO2 extracts to IEC-6 cells. The short-chain fatty acids may be the active ingredient to promote cell proliferation， while paeonol may be the cytotoxic active ingredient.

Objective To develop and explore the clinical application effect of orthopedic protective trestle for post-operative patients with lower limb fracture.Methods Totally 60 patients with lower limb fracture that hospitalized in Osteopathic Department from March to June 2015 were randomly divided into the observation group(n=30) and the control group(n=30). Patients in the control group were given Thomas trestle for the fixation while patients in the observation group were given orthopedic protective trestle for the fixation. The pain score, average hospitalization days and patients′satisfaction degree in two groups were compared. Results The pain score of patients in the observation group after 48 h was (3.50±0.63) points, which was significantly lower than the score in the control group (5.53±0.82) points (t=10.78,P<0.05);the average hospitalization days in the observation group was (16.47±2.68) d, which was significantly lower than that in the control group (22.47± 7.32) d (t=4.22,P<0.05);the satisfaction score of patients in the observation group was (8.80±0.61) points, which was significantly higher than the score in the control group (6.93±1.36) points (t=-6.85,P<0.05). Conclusions Orthopedic protective trestle for patients with lower limb fracture can relieve the pain of patients, reduce the length of average hospitalization days and improve the satisfaction of patients.

Objective To observe the effect of a triptolide-eluting stent(TES)on neointimal hyperplasia in response to vascular injury,inflammation and safety to prevent restenosis after angioplasty.Method Twelve pigs were randomly divided into three groups and received either a bare metal stent(BMS),a sirolimus-eluting stent (SES)or a TES.Each pig was treated with antiplatelet drugs after angioplasty.Biochemistry,vascular morphometry,histopathology and immunohistochemistry were analyzed at 12 weeks after angioplasty.Results The injury scores of the blood vessel were similar in all three groups.There were no differences in minimal lumen diameter or lumen area between the TES[(5.13 ±0.46)mm2;(2.65 ± 0.21)mm]and SES[(5.01±0.54)mm2;(2.65±0.25)mm]groups,but they were significantly(P＜0.01)larger than those in the BMS group[(3.76±0.61)mm2;(2.15 ±0.18)mm].The neointimal area in the TES group was smaller than that in the BMS group,but was similar to that in the SES group.The expression of VEGF,ICAM-1 and α-actinin were significantly lower in the TES group than in the BMS group.In all groups,the proliferation on both edges of the stents was insignificant.No toxicity was found in the TES group.Conclusions TES inhibits neointimal proliferation and the expression of inflammatory factors in pigs.In this study,TES safely and effectively prevented restenosis for 12 weeks.

Objective To screen the MYBPC3 gene mutations in Han Chinese patients with hypertrophic cardiomyopathy ( HCM ). Methods Sixty-six patients with HCM were enrolled for the study. The exons in the functional regions of MYBPC3 were amplified with PCR and the products were sequenced. Results Four novel mutations and four common polymorphisms were identified in this patient cohort. A Lys301fs mutation in exon10 was evidenced in a H30, and when he was 47 years old, he had the chest tightness, shortness of breath with septal hypertrophy of 18. 7mm; a Asp463stop mutation in exonl7 was detected in a H48, he was 24 years old 24-year-old when a medical examination showed ventricular septal hypertrophy of 15.4 mm; both Gly523Arg mutation in exonl8 and Tyr847His mutation in exon26 were found in a H53 with onset age 36 years old, feeling chest tightness after excise and his ventricular septal hypertrophy was 27 mm that time. MYBPC3 mutatons occurred in 4. 5% patients in this cohort. These mutations were not found in 100 non-HCM control patients. Conclusion MYBPC3 mutation is presented in a small portion of Han Chinese patients with HCM.