Objective To establish a mouse model of chronic gratt-versus-host disease (GVHD) after allogeneic hematopoietic stem cell transplantation using chemotherapy preconditioning regimens. Methods The donor mouse was BALB/ cH-2kd and the recipients mouse was C57BL/6H-2kb. The pretreatment of recipient mouse was performed using different doses of chemotherapy drugs, busulfan (BU) 20 mg/(kg • d) ×4 d + cyclophosphamide (CTX) 150 mg/(kg • d) × 2 d, or BU 20 mg/ (kg • d) ×4 d + CTX 100 mg/(kg • d) × 2 d, and then the recipient mouse was injected with different doses of spleen cells (6×107 or 4 × 107 cells) and/or same dose of bone marrow cells (2× 107 cells) to establish the model of chronic GVHD. Hematopoietic chimerism analysis, clinical score and histopathology were used to evaluate the model. Results The high level of recipient-donor mixture chimera was established by pretreated by BU 20 mg/ (kg • d) ×4 d + CTX 150 mg/ (kg • d) × 2d and 2× 107 bone marrow mononuclear cells +6×107 spleen mononuclear cells . The onset of GVHD was concentrated at the 30-90 d after injecting spleen cells of the donor mouse. The clinical scores and incidence rate of chronic GVHD were significantly increased in the large numbers of cells and high doses of chemotherapy transplantation group compared with those in smaller numbers of cells and lower doses of chemotherapy groups (P<0.05). Pathological changes such as cellular and structural abnormalities and inflammatory cell infiltration were noted in the intestines, liver, skin and spleen of the model mice. Conclusion Bone marrow mononuclear cells (2×107 cells) + spleenmononuclear cells (6 × 107 or 4 × 107 cells) can be used to establish a stable chronic GVHD mouse modll via preconditioning with BU and CTX, which paves a way for clinical treatment of chronic GVHD.

Objective To evaluate the efficacy and safety of ustekinumab in the therapy of plaque psoriasis.
Methods Literatures published up to November 2013 were collected from Cochrane library, MEDLINE, and PubMed which were related with ustekinumab for plaque psoriasis. The efficacy was estimated using relative risk of Psoriasis Area and Severity Index (PASI) 75 response rate at the week 12 endpoint in clinical trials, and adverse effects were also analyzed. Meta-analysis was carried out by using Review Manager 5.1.
Results Six randomized control trials consistent with the inclusion criteria were selected and reviewed. Ustekinumab 45 mg group and 90 mg group could get better therapeutic effect compared with the placebo group (all P<0.00001). Furthermore, ustekinumab 90 mg group was more effective than ustekinumab 45 mg group (P=0.01). Adverse effects in the 6 trials were mentioned including headache, upper respiratory tract infection, nasopharyngtis, infection, serious infection, cardiovascular events, and malignant tumors. There were no statistically significant differences of these adverse effects among three groups (all P>0.05), except that infection rate in ustekinumab 45 mg group was higher than the placebo group (P=0.02).
Conclusions Ustekinumab is an effective and safe therapeutic method for plaque psoriasis. However, further longer time analysis of safety is needed.

Objective:To observe the clinical effect of mild moxibustion for knee osteoarthritis (KOA) and to explore the role of mild moxibustion in relieving pain,reducing stiffness and improving joint dysfunction in patients with KOA.Methods:Eighty patients with KOA were randomly allocated into either a moxibustion group or a medication group by the random number table,with 40 cases in each group.The moxibustion group used mild moxibustion at Neixiyan (EX-LE 5),Dubi (ST 35),Xuehai (SP 10) and Liangqiu (ST 34),30 min each time,3 times a week;the medication group was given celecoxib capsule (celebrex),0.2 g each time,once a day.Both groups were treated for 4 weeks.The visual analog scale (VAS) and Western Ontario and McMaster Universities osteoarthritis index (WOMAC)scores were evaluated before and after treatment.The efficacy of the two groups was compared after treatment.Results:After treatment,the overall efficacy of the moxibustion group was significantly different from that of the medication group (P＜0.05).The VAS and WOMAC scores of the two groups were lower than those before treatment (both P＜0.01).The changes in the VAS and WOMAC scores after treatment in the moxibustion group were significantly different from those in the medication group (both P＜0.05).After treatment,in single item of WOMAC,the changes in pain and joint dysfunction in the moxibustion group were more statistically significant than those in the medication group (both P＜0.05).Conclusion:Mild moxibustion and oral celebrex can reduce the VAS and WOMAC scores of patients with KOA.Mild moxibustion is superior to oral celebrex in relieving pain and improving joint function.

Abstract: Objective　To explore the correlation between monocyte to high-density lipoprotein cholesterol ratio (MHR) and insulin resistance (IR) in male patients with type 2 diabetes mellitus (T2DM) combined with metabolic-related fatty liver disease (MAFLD). Methods A total of 454 male patients with T2DM combined with MAFLD in National Metabolic Management Center (MMC) of the Affiliated Hospital of Jiangsu University from May 2018 to July 2020 were enrolled. The general clinical data of subjects were collected, blood routine and biochemical indexes were tested, homeostasis model insulin resistance index (HOMA-IR) was calculated, visceral fat area (VFA) and subcutaneous fat area (SFA) were measured. Accordingtothe MHR quartile, patients were divided into group Q1 (MHR≤0.38), group Q2 (0.380.64) to compare the differences in measured indicators above. In addition, patients were divided into two groups according to HOMA-IR, HOMA-IR<2.5 and HOMA-IR≥2.5, and the differences in MHR were compared. Results The patients were divided into four groups according to MHR:group Q1 (n=115), group Q2 (n=110), group Q3 (n=120) and group Q4 (n=109). Fasting insulin (FINS) were respectively 6.17(4.20,9.76), 7.73(4.94,10.66), 8.92(5.32,11.33) and 9.13(5.25,12.27) mU/L, 2-hour postprandial insulin were 22.75(12.87,39.59), 27.55(16.44,39.77), 30.98(17.46,43.11) and 31.28(18.54,45.92) U/L. HOMA-IR were 3.12(1.63,4.25), 3.72(2.26,4.66), 3.87(2.48,5.44) and 3.95(2.42,5.31). Neutrophil (Neu) were 3.10(2.60,3.70), 3.20(2.50,3.93), 3.60(2.80,4.28), 4.20(3.30,5.00)×109/L. Subcutaneous fat area (SFA) were (181.27±53.60), (192.64±62.41), (199.53±61.40) and (203.69±71.51) cm2. They all increased gradually. However, the levels of high-density lipoprotein cholesterol (HDL-c) [1.18(1.06,1.35), 1.02(0.86,1.17), 0.96(0.80,1.03) and 0.80(0.69,0.92) mmol/L] and low-density lipoprotein cholesterol (LDL-c) [(3.00±0.79), (2.76±0.83), (2.67±0.85) and (2.59±0.92) mmol/L] decreased gradually. Pearson's or Spearman's correlation analysis showed that MHR was positively correlated with FINS, 2-hour postprandial insulin (2hINS), HOMA-IR, VFA and SFA (r=0.190, 0.153, 0.184, 0.114, 0.127, P<0.05). The coronary heart disease history, systolic blood pressure,diastolic blood pressure,fasting plasmaglucose (FPG), FINS, alanine aminotransferase (ALT), aspartate aminotransferase (AST), blood uric acid (Ur), body mass index (BMI), VFA, SFA and MHR of patients in group HOMA-IR≥2.5 were higher than group HOMA-IR<2.5 (P<0.05). Conclusion　MHR is positively correlated with IR in male patients with T2DM combined with MAFLD, and as MHR increases, the degree of IR is higher.

OBJECTIVETo observe the efficacy of He-wei Jie-gu (Chinese characters) decoction on femoral intertrochanteric fractures after internal fixation in elderly.

METHODSFrom January 2010 to January 2012, 90 elderly patients with osteoporotic intertrochanteric fracture were randomly divided into two groups. The patients in experimental group were treated with open reduction and internal fixation, He-wei Jie-gu decoction and Caltrate D in oral, including 18 males and 27 females with an average age of (77.8±11.3) years old ranging from 65 to 89 years old;the time from injured to operation was 2 to 9 days with an average of (6.6±2.1) days; according to Evans-Jensen classification, 17 cases were in type I, 8 in type II, 15 in type III, 3 in type IV, 2 in type V. Meanwhile, the patients in control group were treated with open reduction and internal fixation, Caltrate D in oral, including 16 males and 29 females with an average age of (81.2±9.6) years old ranging from 67 to 91 years old; the time from injury to operation was 2 to 10 days with an average of (6.8±2.6) days; according to Evans-Jensen classification, 15 cases were in type I, 10 in type II, 14 in type III, 4 in type W, 2 in type V. The VAS score, hip BMD value before and after treatment, fracture healing and the Harris function score after operation were recorded and compared.

RESULTSAll patients were followed up from 3 to 9 months with an average of (5.6±2.8) months. Compared with the control group, the VAS score of the experimental group had an obviously reduction (F=98.138, P<0.01) after treatment; the hip BMD value at 3 months after treatment in the experimental group was higher than that of the control group (P<0.05); the fracture healing rate of the experimental group was higher than that of the control group (P<0.05); Harris score in 3 months of follow-up in the experimental group was higher than that of the control group (t=2.542, P<0.05); all the differences between two groups were statistically significant (P<0.05).

CONCLUSIONHe-wei Jie-gu decoction can reduce postoperative pain and increase bone mineral density after internal fixation of femoral intertrochanteric fracture in elderly, promote the fracture healing and functional recovery in the hip joint.

Aged ; Aged, 80 and over ; Bone Density ; Combined Modality Therapy ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Femur ; injuries ; surgery ; Fracture Fixation, Internal ; Hip Fractures ; surgery ; Humans ; Male ; Osteoporotic Fractures ; drug therapy ; physiopathology ; surgery ; Pain, Postoperative ; drug therapy ; physiopathology

Objective To observe the formation of Staphylococcus aureus biofilm and the inhibitory and dispersive effects of betaine on the biofilm.Methods The inhibitory and dispersive effects of 0.1％ betaine on the biofilm from 20 strains of Staphylococcus aureus were examined by crystal violet assay.Results All the 20 strains of Staphylococcus aureus formed biofilm.The biofilm of methicillinsensitive Staphylococcus aureus (MSSA) was formed in 24 hours with peak value of absorbance (A590 nm) (1.99 ± 0.53).The biofilm of methicillin-resistant Staphylococcus atureus(MRSA) was formed in 48 hours with peak value of absorbance(A590 nm) (1.13 ±0.47).After adding betaine,the absorbance(A590 nm) of MSSA biofilm fell down to(1.74 ± 0.61) in 24 hours,while the absorbance(A590 nm) of MRSA biofilm fell down to(0.40 ± 0.12) in 48 hours,which was significantly reduced compared with the controls (t =2.43,5.84,P ＜ 0.05 respectively).When adding betaine after the biofilm formed,the absorbancies (A590 nm) of both MSSA and MRSA showed no significant difference compared with the controls (P ＞ 0.05).Conclusion Betaine could inhibit biofilm formation of Staphylococcus aureus at concentration of 0.1％,but it could not disperse the mature biofilm of Staphylococcus aureus.

As the increasing population aging,the incidence of age-related macular degeneration is increasing,blinding rate also increasing,so it is very important for the early diagnosis and treatment of age-related macular degeneration.There are many methods to check the aging macular degeneration,such as fundus angiography,optical coherence tomography (OCT),visual field and multifocal electroretinogram (mfERG).In recent years,many emerging ophthalmic methods have emerged and are gradually applied to clinical diagnosis,including optic coherence tomography angiography (OCTA).The function of these methods has its unique advantages,but there are also limitations.This paper will review these existing methods.

Lipase gene from Penicillium expansum(lip07) was cloned and over-expressed in Pichia pastoris.a random mutant named ep8,which contained a single amino acid substitution,was obtained by using the lip07 as an error-prone PCR template in previous study.ep8 shows higher thermostability than that of lip07,To further improve the thermostability of the lipase,the Lys of wild-type(lip07) and mutant(ep8) in 202 were substituted by Ala using the Overlap extension PCR technique respectively.The mutant genes(lip07-K202A and ep8-K202A) were subcloned into pAO815,and then transformed into the Pichia pastoris GS115 for extracelluar expression,respectively.15% SDS-PAGE analysis indicated that the molecular mass of PEL-ep8-K202A and PEL-lip07-K202A are both about 28kDa,which is same with the wild-type lipase.The Tm of PEL-ep8-K202A is 41.66℃,2.63℃ higher than that of the wild-type(39.03℃) and 1.21℃ higher than the random mutant(PEL-ep8:40.45℃);the Tm of single mutant(PEL-lip07-K202A) is 37.08℃,2℃ lower than that of the wild-type lipase.

AIM:Targeting of focal adhesion kinase (FAK) gene,we aim to construct FAK shRNA lentiviral vector and to identify its function on the growth of leukemic cells.METHODS:FAK shRNA was chemically synthesized,and inserted into a GFP-lentiviral plasmid through molecular biology methods.After packaged and concentrated,the lentiviral-FAK-shRNA-vector was transduced into a human leukemic cell line.FAK gene expression was detected by reverse transcriptional PCR and Western blotting.Cell apoptosis was measured by Annexin V labeling.RESULTS:The results showed that FAK shRNA was successfully inserted into the lentival vector,and the infection efficiency varied from 10% to 25%.Compared to the control vector (lentival vector without FAK shRNA),FAK shRNA inhibited the expression of FAK mRNA and protein by 40% and 70%,respectively.Moreover,the results of apoptosis experiment showed that the percentages of Annexin V+ cells in control vector group and FAK shRNA group were (4.19 ? 0.36) % and (8.48 ? 0.58) % respectively,the difference was statistically significant (P

?Age-related macular degeneration ( AMD) is one of the main leading causes of irreversible damage eyesight over the 50 years old people. Genetic factors play an important role in the occurrence and development of AMD. ln recent years, the studies found that complement factor H ( CFH) gene has obvious correlation with the incidence of AMD. ln this article, we reviewed the researches on the CFH in AMD.