By means of preparative HPTLC and column chromatography over silica gel and Sephadex LH-20, nine diterpenoids were isolated and purified from the whole plants of Scutellaria strigillosa. Based on the physico-chemical properties and spectral data, their structures were elucidated as: 6-O-acetyl-7-O-nicotinoylscutebarbatine G(1), 6-O-nicotinoyl-7-O-acetylscutebarbatine G(2), 6,7-di-O-nicotinoylscutebarbatine G(3), scutebarbatine K(4), scutebarbatine B(5), 6-O-acetylscutehenanine A(6), 6-O-nicotinoylbarba- tin A(7), 6,7-di-O-acetoxylbarbatin A(8), scutebarbatine F(9). Compound 1 is a new diterpenoid, and compounds 2-9 were isolated from Scutellaria strigillosa for the first time.
Diterpenes ; chemistry ; Drugs, Chinese Herbal ; chemistry ; Magnetic Resonance Spectroscopy ; Molecular Structure ; Scutellaria ; chemistry ; Spectrometry, Mass, Electrospray Ionization

Objective To investigate the clinical efficacy of Qishen Xiaodian Decoction combined with laser acupoint irradiation for treatment of recurrent Hen?ch-Sch?nlein purpura (HSP) in children. Methods Totally 120 cases of children with recurrent HSP were divided into treatment group and control group according to the digital random table method, with 60 cases in each group. The control group was given anti-allergy, hormones, immunosuppressive agents and other conventional treatment for 4 weeks, while the treatment group was treated with Qishen Xiaodian Decoction based on conventional treatment, 1 dose each day, morning and night (age 4–5 take 1/3 dose, age >5–10 take the half dose, and age >10–14 take the whole dose), for 4 weeks. Zusanli, Xuehai, and Sanyinjiao acupoints were under laser vertical irradiation, 12 min for each time, once a day for 2 weeks. The disappearing time of the main symptoms and the total effective rate of the two groups were compared. Peripheral blood contents of Th17 and Treg cells and serum interleukin-17 (IL-17), and transforming growth factor β1 (TGF-β1) level before and after the treatment were observed. The recurrence rate in 6 months and in 12 months of the two groups were compared. Results The disappearing time of rash, abdominal pain and joint swelling pain and kidney damage of the treatment group were less than those of the control group (P<0.01). The total effective rate was 95％(57/60) in treatment group, significantly higher than control group 80％ (48/60), with statistical significance (P<0.05). Compared with before treatment, Th17 cell content, serum IL-17 and TGF-β1 decreased, and Treg cell content increased after treatment of the two groups (P<0.01). Compared with the control group, contents of Th17 cell and serum IL-17 of the treatment group decreased (P<0.01). The recurrence rate was lower than that of the control group in 6 and 12 months after treatment, with statistical significance (P<0.05). Conclusion Based on the conventional western medicine treatment, Qishen Xiaodian Decoction combined with laser acupoint irradiation on recurrent HSP in children can quickly relieve symptoms and reduce the recurrence rate.

OBJECTIVETo investigate the effect of apoptosis of CD4+ CD25+ regulatory T cells (Tregs) on proliferation as well as secretory function of effector T cells (Teff) and potential influence of Xuebijing injection on them in septic rats.

METHODSA sepsis model was reproduced by cecal ligation puncture (CLP), and Wistar rats were randomly divided into the control group (n = 8), sham-operated group (n = 8), CLP group (n = 8), and Xuebijing injection treatment group (n = 8). CD4+ CD25+ Tregs in each group were separated by immunomagnetic beads isolate system on day 3, the apoptosis rate, expression of forkhead/winged helix transcription factor p3 (Foxp3) and cytotoxic T-lymphocyte-associated antigen 4 (CTLA-4) on Tregs were analyzed by flow cytometry, and secretion levels of interleukin (IL)-10 from Tregs were measured by ELISA. Following co-culture of CD4+ CD25+ Treg with CD4+ CD25- T cells (1:1) for 68 hours, proliferative activity of Teff was determined by MTT, and IL-2/sIL-2R alpha levels were measured by ELISA.

RESULTSThe apoptosis rate of Tregs in control group was 12.03% +/- 0.89%, which was not significantly different from sham-operated group 9.48% +/- 2.17%. The apoptosis rate of Tregs in CLP group 5.87% +/- 0.44% was lower than that in control group (P < 0.01), and treatment with Xuebijing injection markedly enhanced the apoptosis of Tregs 27.29% +/- 2.48%. Compared to CLP group, expression of Foxp3, CTLA-4, and the secretion of IL-10 of Treg were significantly lowered in Xuebijing injection group (all P < 0.01). The Teff proliferative activity in response to ConA, and IL-2 levels of Teff in CLP group were significantly suppressed compared with control group (P < 0.01), and secretion of sIL-2R alpha in the supernatants was much higher than that of the control group. In comparison to the CLP group, inhibitory rate of Teff proliferative activity and the sIL-2R alpha levels were significantly decreased, while the secretion of IL-2 was increased in Xuebijing injection group (P <0.01).

CONCLUSIONCD4+ CD25+ Tregs could markedly upregulate the suppressive function on Teff in sepsis, and treatment with Xuebijing injection effectively enhanced apoptosis of Tregs, thereby down-regulating the suppression on Teff.

Animals ; Apoptosis ; Cell Proliferation ; Disease Models, Animal ; Drugs, Chinese Herbal ; therapeutic use ; Male ; Rats ; Rats, Wistar ; Sepsis ; drug therapy ; immunology ; T-Lymphocytes ; immunology ; pathology ; T-Lymphocytes, Regulatory ; immunology ; pathology

OBJECTIVETo retrospectively study medium-term follow-up outcomes of total hip arthroplasty (THA) for patients with ankylosing spondylitis (AS).

METHODSFrom January 2000 to December 2008, 67 patients (88 hip joints) with AS were treated with all ceramic interface THA. And 55 patients (74 hips) were finally followed up. Among them, there were 30 males and 25 females, with an average age of 32.6 (ranged 19 to 58) years old. Sixty-one hips were treated with biological prosthesis and 13 hips were treated with hybrid prosthesis. Fifty-five patients were followed up at least 5 years, with an average of (75.2 +/- 8.6) months. Clinical symptoms and radiography information were evaluated after follow-up.

RESULTSHarris hip score were significantly improved from 30.8 +/- 7.0 preoperatively to 85.2 +/- 5.5 at the last follow-up (P<0.01). The hip movement range increased from (21.2 +/- 8.5) degrees preoperatively to (142.0 +/- 10.2) degrees postoperatively (P<0.01). The 5-year survival of prosthesis was 95.9%. One patient were renovated because of internal wall broken caused by injury, 1 was renovated for infection, 1 was renovated for fracture arround femoral stem prostheses, and 1 was treated with conservative treatment by dislocation. Three cases with abnormal sound were cured with non-operation. 7 cases with heterotopic ossification were not treated, 2 cases with thigh pain received conservative treatment. Bone dissolve around prosthesis, loose and sink of femur and acetabulum prosthesis were occurred in other cases.

CONCLUSIONTHA for the treatment of AS is a reliable method, which has a satisfied medium-term follow-up outcomes.

Adult ; Arthroplasty, Replacement, Hip ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Spondylitis, Ankylosing ; surgery ; Treatment Outcome ; Young Adult

OBJECTIVETo investigate the enhancing effect of Chinese medicine-Xuebijing injection on lipopolysaccharide (LPS) -induced apoptosis of CD4+ CD25+ regulatory T cells (Tregs) and polarization of helper T cells (Th).

METHODSCD4+ CD25+ Tregs collected from rat spleen in vitro by immunomagnetic beads assay were divided into the control group, anti-CD3/CD28 group, anti-CD3/CD28 + LPS group, anti-CD3/CD28 + "Xuebijing injection" group and anti-CD3/CD28 + LPS + "Xuebijing injection" group. Tregs apoptosis rate and expression of winged helix transcription factor (Foxp3) in Tregs were detected by flow cytometry on 3rd post culture day. CD4+ CD25- T cells were co-cultured with CD4+ CD25- Tregs (1:1) for 68 hours with canavalin A stimulation. Interferon gamma (gamma-IFN), interleukin (IL)-4 and IL-17 in supernatants, which respectively was secreted by Th1, Th2 and Th17, were measured by ELISA.

RESULTSTregs apoptosis rate of anti-CD3/CD28 + LPS + "Xuebijing injection" group (45.1 +/- 2.7%) was significantly higher than that of anti-CD3/CD28 + LPS group (29.4 +/- 1.6%, P < 0.01). Meanwhile, Foxp3 expressions in Tregs in above 2 groups were 95 +/- 9 and 140 +/- 18 respectively, showing statistically significant difference between them (P < 0.01). Gamma-IFN levels secreted in anti-CD3/CD28 + LPS + "Xuebijing injection" group were significantly higher than those in anti-CD3/CD28 + LPS group (P < 0.01), while IL-4 levels had an opposite tendency compared with gamma-IFN (P < 0.05), resulting in a marked increase in the ra- tio of gamma-IFN/IL-4 in anti-CD3/CD28 + LPS + "Xuebijing injection" group (P < 0.01). In anti-CD3/ CD28 + "Xuebijing injection" group, IL-17 secretion levels were significantly decreased compared with anti-CD3/CD28 group (P < 0.05).

CONCLUSIONSActivation of CD4+ CD25+ Tregs induced by LPS may mediate Th1 shift to Th2 response. "Xuebijing injection" can effectively regulate immune function of T cells, increase the LPS-induced apoptosis of CD4+ CD25+ Tregs as well as enhance the polarization of Th2 to Th1, thereby abating the suppressive state of cell-mediated immunity.

Animals ; Apoptosis ; Drugs, Chinese Herbal ; pharmacology ; Endotoxins ; Lipopolysaccharides ; Male ; Rats ; Rats, Wistar ; T-Lymphocytes, Helper-Inducer ; drug effects ; immunology ; T-Lymphocytes, Regulatory ; drug effects ; immunology

In this paper, we studied the relationship between the prostaglandin F(2alpha) (PGF(2alpha))-induced cardiac hypertrophy and calcineurin (CaN) signal transduction pathway in vivo and in vitro. Male Sprague-Dawley rats were given a single i.p. injection with monocrotaline (MCT) (60 mg/kg) and then given orally with celecoxib (20 mg/kg) or vehicle once a day for 14 d before (from d 1 to d 14) or after (from d 15 to d 28) right ventricular hypertrophy (RVH) was formed. Body weight (BW), right ventricular weight (RV), left ventricular with septum weight (LV), as well as lung weight were determined. RVH index (RVHI=RV/LV), RV/BW, and lung weight/BW were calculated and histological changes were observed with transmission electron microscope. PGF(2alpha) level, atrial natriuretic peptide (ANP) and CaN mRNA expressions, expression of CaN and its downstream effectors, NFAT(3) and GATA(4) protein were assayed by EIA kit, RT-PCR, and Western blotting, respectively. The cardiomyocyte hypertrophy in primary culture induced by PGF(2alpha) (0.1 micromol/L) was evaluated by measuring the cell diameter, protein content, and ANP mRNA as well as CaN mRNA expressions. It was found that 14 d or 28 d after MCT was given, the RVHI, RV/BW, and lung weight/BW were significantly increased by 47%, 53% and 118%, and by 64%, 94% and 156%, respectively; at the same time PGF(2alpha) levels in RV tissue were increased by 44% and by 51% with increasing RVHI, and elevated expressions of ANP and CaN mRNA, as well as CaN, NFAT(3) and GATA(4) proteins in a positive correlation manner. Furthermore, some histological injuries were found in RV tissue. Celecoxib, a cyclooxygenase inhibitor, obviously blunted the elevation of RVHI, RV/BW, and lung weight/BW no matter it was given before or after RVH. In vitro experiments showed that 0.1 micromol/L PGF(2alpha) significantly increased the cardiomyocyte diameter and protein content, and promoted ANP and CaN mRNA expressions, which was blocked by cyclosporin A, a CaN inhibitor. Our results indicate that PGF(2alpha) may be involved in cardiac hypertrophy induced by MCT in rats through CaN signal transduction pathway.
Animals ; Calcineurin ; genetics ; metabolism ; physiology ; Cells, Cultured ; Dinoprost ; metabolism ; physiology ; Hypertrophy, Right Ventricular ; chemically induced ; metabolism ; physiopathology ; Male ; Monocrotaline ; Myocytes, Cardiac ; metabolism ; pathology ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; physiology

OBJECTIVETo establish a novel method for the multiplex analysis of the methylation and single nucleotide polymorphism (SNP).

METHODSThe imprinted SNP rs220028 was chosen as a model. Genomic DNA, after being digested with methylation sensitive restriction enzyme, were typed by mutagenically separated PCR (MS-PCR). The polymorphism of restriction site was excluded by PCR-RFLP.

RESULTSBy post-digestion MS-PCR, the methylated allele was detected selectively, the maternal origin of which was confirmed by pedigree analysis; A=0.5085, G=0.4915,PIC=0.3749.

CONCLUSIONThe multiplex analysis of methylation markers and SNP can be achieved by post-digestion MS-PCR. The imprinted SNP locus rs220028 is a potentially useful marker in screening Prader-Willi/Angelman syndrome.

DNA Methylation ; DNA Restriction Enzymes ; metabolism ; Genetic Markers ; genetics ; Humans ; Polymerase Chain Reaction ; methods ; Polymorphism, Single Nucleotide

OBJECTIVE: To establish a new method for single nucleotide polymorphism (SNP) typing based on allele specific PCR: fragment length discrepant allele specific PCR (FLDAS-PCR), and study the influence on specific extension by introducing a mismatch at the third or fourth 3'-terminal base of allele specific primers. METHODS: For SNP loci rs759117 and rs760887, two allele specific forward primers, with different length and a mismatch introduced at the third or fourth 3'-terminal base, and a public reverse primer were designed for SNP typing. The genotyping of SNP was determined by the two allele specific fragments different in size after polyacrylamide gel and silver staining. RESULTS: The different homozygote genotypes comprised a single band with different size respectively, and the heterozygote genotypes comprised two bands. Typing results were completely consistent with those by direct sequencing. Non-specific primer extension was decreased remarkably after introducing a mismatch at the third or fourth 3'-terminal base of allele specific primers, and the stringency of PCR reaction was cut down. CONCLUSION FLDAS-PCR is a simple, rapid and efficient new method for SNP typing. During FLDAS-PCR, specific primers with a mismatch at the third or fourth 3'-terminal base have more power to identify two alleles.
Alleles ; Base Pair Mismatch/genetics* ; DNA/genetics* ; DNA Primers ; Electrophoresis, Polyacrylamide Gel ; Genotype ; Humans ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length ; Polymorphism, Single Nucleotide ; Sensitivity and Specificity ; Sequence Analysis, DNA/methods*

BACKGROUNDTotal knee arthroplasties (TKR) combined with the concept of reduced trauma to tissue has been performed by many doctors. The aim of this study was to retrospectively assess the early results of a group of TKR with a mid-vastus approach, which was characterized as no patellar eversion, no disruption of suprapatellar pouch and extensor mechanism, and to compare the outcome with conventional operative techniques.

METHODSA total of 59 patients (67 knees) were followed. All patients received the same prosthesis of Genesis II posterior-stabilized total knees. Of them, 29 consecutive patients (34 knees) had a mid-vastus approach and were operated on with less invasive instruments and techniques. The mean follow-up duration was 11.6 months. Clinical evaluations were performed according to the Hospital for Special Surgery scores; radiographic assessment followed the guidelines of the Knee Society. Postoperative recovery of quadriceps strength and the extensor mechanism was also evaluated.

RESULTSNo prosthetic loosening or anterior knee pain was found at the latest follow-up; 1 patient had a superficial infection and postoperative stiffness of the knee who undertook a debridement and manipulation several months later. The angulations of tibial osteotomy were within normal range. The average preoperative and postoperative Hospital for Special Surgery scores were 57.9 points and 86.1 points respectively. The mean postoperative range of motion was 113.5 degrees . Most patients regained their quadriceps strength at the third or fourth month postoperatively.

CONCLUSIONSThe overall early results from using the mid-vastus approach were comparable with that of using a standard approach, and the exposure did not affect the accuracy of the tibial cut. The muscle maximal contraction strength recovered gradually after operation. The approach was safe and patients of this group obtained satisfactory outcomes.

Adult ; Aged ; Aged, 80 and over ; Arthroplasty, Replacement, Knee ; methods ; Female ; Humans ; Male ; Middle Aged ; Range of Motion, Articular

Solid organ transplant recipients are at increased risk for Aspergillus infections. However, the cases of Aspergillus spondylodiscitis are rare and mostly resulted from the hematogenous spread of invasive pulmonary Aspergillosis. Here, we report a case of primary spondylodiscitis in a liver transplant recipient. Six months after transplantation, a chronic and progressive lumbar back pain was presented. The patient had no fever and the white blood cell count was normal. High plasma (1→3)-beta-d-glucan (BDG) level was detected at the time of back pain. The pathogen was Aspergillus flavus. Clinical and radiological healing was achieved through posterior only debridement and voriconazole therapy.
Adult ; Aspergillosis ; blood ; diagnosis ; etiology ; Discitis ; blood ; diagnosis ; etiology ; Humans ; Liver Transplantation ; adverse effects ; Male