OBJECTIVETo develop an HPLC method for determination of gallic acid, hydroxysafflor yellow A, cinnamic aldehyde and piperine in Tibetan medicine Dangzuo, and to compare the content of four active components in Dangzuo of different Tibetan regions.

METHODThe separation was carried out on a Waters XTerra RP-C18 column ( 4.6 mm x 250 mm, 5 microm). The mobile phases were methanol and water, all contained 0.1% glacial acetic acid, for gradient elution. The gradient program was as follows: 0-22.5 min, methanol was changed from 5% to 50%; 22.5-40 min, changed to 80% 80:20. The flow rate was 1.0 mL x min(-1). The detection wavelength was 270 nm. The reference wavelength was 500 nm.

RESULTThe linear ranges of gallic acid, hydroxysafflor yellow A, cinnamic aldehyde and piperine were 0.040-0.640 microg (r = 0.999 8), 0.090-1.440 microg (r = 0.999 9), 0.031-0.500 microg (r = 0.999 9 ) and 0.092-41.477 microg (r = 0.998 9), respectively. The average recoveries (n = 6) were 97.42% (RSD 1.9%), 97.55% (RSD 2.9%), 98.69% (RSD 0.96%) and 96.72% (RSD 4.0%), respectively. The content ranges of gallic acid, hydroxysafflor yellow A, cinnamic aldehyde and piperine in Dangzuo samples of different Tibetan regions were 0.11341.69 mg x g(-1), 0.889-1.51 mg x g(-1), 0.000-40.606 mg x g(-1) and 1.96-2.73 mg x g(-1), respectively.

CONCLUSIONThe method is a simple and effective for quality control of Tibetan medicine Dangzuo.

Acrolein ; analogs & derivatives ; analysis ; isolation & purification ; Alkaloids ; analysis ; isolation & purification ; Benzodioxoles ; analysis ; isolation & purification ; Chalcone ; analogs & derivatives ; analysis ; isolation & purification ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; Gallic Acid ; analysis ; isolation & purification ; Medicine, Tibetan Traditional ; Piperidines ; analysis ; isolation & purification ; Plant Components, Aerial ; chemistry ; Plant Extracts ; analysis ; isolation & purification ; Polyunsaturated Alkamides ; analysis ; isolation & purification ; Quality Control ; Quinones ; analysis ; isolation & purification ; Reference Standards ; Spectrophotometry, Ultraviolet ; methods

In the present study, through a functional strategy, a metagenome library of the marine microbes from the surface water of the South China Sea was screened for beta-glucosidase and six positive clones were obtained. One of these clones, pSB47B2, was subcloned and further analysed in sequence. The result showed that there was an open reading frame for a novel beta-glucosidase, which was nominated as bgl1B. Using pET22b(+) as vector and Escherichia coli BL21(DE3) as host, Bgl1B was overexpressed recombinantly with high yield obtained and substantial enzymatic activity detected. The recombinant protein (rBgllB) was purified by Ni-NTA affinity chromatography and further biochemically characterized. The results indicated that, with pNPG as substrate, the optimum pH and temperature for the hydrolytic activity of rBgl1B were about 6.5 and 40 degrees C respectively. Under the optimum conditions, rBgl1B hydrolyzed pNPG with an activity up to 39.7 U/mg, Km and Vmax being 0.288 mmol/L and 36.9 micromol/min respectively. In addition, rBgl1B could also hydrolyze cellobiose, with a Km of 0.173 mmol/L and a Vmax of 35 micromol/min. However, we did not detect evident hydrolytic activity of rBgl1B to lactose, maltose, sucrose, and CMC. The enzymatic activity of rBgl1B to pNPG was stimulated to certain degrees by low concentration of Ca2+ or Mn2+, whereas it exhibited significant tolerance against high Na+. Distinguished from most of the beta-glucosidases derived from fungi, which display the highest activities under acidic conditions, rBgl1B exhibited relatively higher activity and stability at pH between 7.0 and 9.0.
Amino Acid Sequence ; Cloning, Molecular ; Enzyme Stability ; Escherichia coli ; genetics ; metabolism ; Metagenome ; genetics ; Metagenomics ; methods ; Molecular Sequence Data ; Recombinant Proteins ; biosynthesis ; genetics ; isolation & purification ; Seawater ; microbiology ; beta-Glucosidase ; biosynthesis ; genetics

Objective To investigate the clinical effect of percutaneous hollow screw combined with tension band internal fixation for comminuted patellar fracture after arthroscopic reduction.Methods 105 patients with comminuted patellar fracture in our hospital from April 2013 to September 2017 were selected and divided into observation group (n =53) and control group (n =52) according to different treatment schemes.The control group was treated with conventional open reduction and Kirschner wire combined with tension band internal fixation,while the observation group was treated with arthroscopic assisted reduction and percutaneous cannulated screw combined with wire tension band internal fixation.The operation status and postoperative fracture healing time of the two groups were compared.One-year follow-up was performed after operation.The incidence of complications,the excellent and good rate of operation,and the preoperative and postoperative pain scores (VAS) and knee were compared.Hospital for special surgery knee score (HSS),Activity of Daily Life Scale (ADL),quality of life score (the Mos 36-item Short Form Health Survey,SF-36) were compared.Results The observation group had fewer fluoroscopy times during operation,shorter incision length and shorter healing time after operation than the control group (P ＜ 0.05);the incidence of complications in the observation group was 3.85％,lower than 20.00％ in the control group,and the excellent and good rate of operation was 96.15％,higher than 82.00％ in the control group (P ＜ 0.05);the VAS scores of the two groups were lower than those of the control group one year after operation,and the scores of HSS,ADL and SF-36 were lower than those of the control group.The preoperative improvement was higher in the observation group than in the control group (P ＜ 0.05).Conclusions Arthroscopic reduction combined with percutaneous hollow screw and tension band internal fixation for patellar comminuted fracture can reduce surgical trauma and accelerate the rehabilitation of patients.It can reduce the incidence of complications,relieve pain,improve knee function,daily living ability and quality of life,and the effect is remarkable.

BACKGROUND:Currently,there is a lack of large sample studies to analyze the bone metabolism level of patients with femoral head necrosis of different etiologies and stages,which is not conducive to the development of better necrosis-promoting repair strategies. OBJECTIVE:To study the bone metabolism of patients with osteonecrosis of the femoral head with different etiologies and Association Research Circulation Osseous(ARCO)stages. METHODS:A retrospective study was performed on 401 patients diagnosed with osteonecrosis of the femoral head as the trial group,and 81 healthy subjects as the control group.The trial group could be divided into three groups according to different etiologies:steroid-induced osteonecrosis of the femoral head,alcoholic osteonecrosis of the femoral head and traumatic osteonecrosis of the femoral head,and were divided into stages Ⅱ/Ⅲ/Ⅳ according to different ARCO stages.Seven bone metabolism-related indicators of all subjects were collected,including bone metabolism-regulating hormone 25-hydroxyvitamin D and bone conversion markers:N-terminal propeptide of type Ⅰ procollagen,degradation product of type Ⅰ collagen,n-terminal middle molecular fragment of osteocalcin,general biochemical markers of bone metabolism:serum calcium,serum phosphorus,serum alkaline phosphatase.The bone metabolism levels of each group were compared and the independent factors associated with osteonecrosis of the femoral head were determined by binary Logistic regression analysis. RESULTS AND CONCLUSION:Compared with the control group,levels of degradation product of type Ⅰ collagen,N-terminal propeptide of type Ⅰ procollagen,n-terminal middle molecular fragment of osteocalcin,serum phosphorus and alkaline phosphatase in the trial group were significantly increased(all P＜0.05).Based on the presence or absence of the disease,according to binary Logistic regression analysis,degradation product of type Ⅰ collagen,N-terminal propeptide of type Ⅰ procollagen,and n-terminal middle molecular fragment of osteocalcin were independent factors associated with osteonecrosis of the femoral head.The levels of degradation product of type Ⅰ collagen and N-terminal propeptide of type Ⅰ procollagen in three groups of patients with different etiologies were higher than normal reference values.The bone metabolism-regulating hormone 25-hydroxyvitamin D and serum calcium in the alcoholic osteonecrosis of the femoral head group were higher than those in the other two groups(P＜0.05).The level of bone metabolism-regulating hormone 25-hydroxyvitamin D in steroid-induced and traumatic osteonecrosis of the femoral head groups was lower than the normal value.There were no significant differences in seven bone metabolism-related indicators in patients with ARCO stages Ⅱ,Ⅲ and Ⅳ osteonecrosis of the femoral head(all P＞0.05),but degradation product of type Ⅰ collagen and N-terminal propeptide of type Ⅰ procollagen in these three groups were higher than normal reference values.Bone metabolism-regulating hormone 25-hydroxyvitamin D in patients with ARCO stage Ⅱ and ARCO stage Ⅳ was lower than the normal reference value.It is concluded that the bone metabolism level of osteonecrosis of the femoral head patients was abnormal.The degradation product of type Ⅰ collagen and N-terminal propeptide of type Ⅰ procollagen of osteonecrosis of the femoral head patients with different etiologies and ARCO stages were all higher than the normal reference value,and they were in a state of high bone turnover.Degradation product of type Ⅰ collagen,N-terminal propeptide of type Ⅰ procollagen and n-terminal middle molecular fragment of osteocalcin may be risk factors for the pathogenesis of osteonecrosis of the femoral head.

OBJECTIVETo study the heat processing technics of Nanhanshuishi.

METHODTo find the best processing technic, the single factor experiments and orthogonal experiments were designed basing on the processing technics summarized by consulting documents, scriptures and investigating some Tibetan hospitals, meanwhile, the content of Ca, Fe, Mn, Zn, and Cu in the processed Nanhanshuishi in single factor experiments and orthogonal experiments were detected.

RESULTThe best processing technic of Nanhanshuishi was as follows: Nanhanshuishi was crashed to 10-20 mm in diameter, the ratio of the weight of Aconiti Kusnezoffii Radix and potassium nitrate was 1: 2, and the boiling time was 3 h.

CONCLUSIONThe work in this article provided a basic processing technic data for clarifying the mechanism of processing and establishing the perfect processing technics of Nanhanshuishi.

Medicine, Tibetan Traditional ; Technology, Pharmaceutical ; Trace Elements ; analysis

Objective To compare the diagnostic efficiency of the thyroid imaging reporting and data system (TIRADS), the Bethesda system for reporting thyroid cytopathology(BSRTC) and BRAFV600E detection, and their combined use in the differentiation between benign and malignant thyroid nodules. Methods One hundred and twenty eight patients with 128 thyroid nodules who were scheduled for ultrasound-guided fine-needle aspiration biopsy (FNAB) were recruited for the study. All of them underwent ultrasound, fine-needle aspiration cytology(FNAC) examination, and BRAFV600E detection. TIRADS and BSRTC systems were adopted to judge the ultrasound and FNAC results. The receiver operating characteristic curve was established to assess the diagnostic value of each method. Results The sensitivity, specificity, and AUC of TIRADS were 74. 3%, 84. 5%, and 0. 794, respectively. BSRTC had higher specificity(98. 3%) and equal sensitivity compared to TIRADS. The sensitivity, specificity, and AUC of BRAFV600E detection were the highest ones among the three methods. Combinations of different methods could increase the diagnostic sensitivity and accuracy. The combination of FNAC and BRAFV600E detection significantly increased the diagnostic efficiency(AUC=0. 984), with sensitivity 98. 6% and specificity 98. 3%. Conclusions The diagnostic value of BRAFV600E detection in the differentiation of benign and malignant thyroid nodules is better than both TIRADS and BSRTC, and the combination of FNAC and BRAFV600E detection reaches the best diagnostic efficiency.

OBJECTIVETo evaluate the effect of preoperative transcatheter arterial chemoembolization(TACE) on apoptosis of hepatocellular carcinoma (HCC) cells.

METHODSA total of 136 patients with HCC underwent liver resection. One to five courses of TACE prior to liver resection were performed in 79 patients (TACE group), in which one to four courses of chemotherapy alone were performed in 11 patients (group A), one to five courses of chemotherapy combined with iodized oil were performed in 33 patients (group B), one to three courses of chemotherapy combined with iodized oil and gelatin sponge were performed in 23 patients group C) and one to three courses of chemotherapy combined with iodized oil, ethanol and gelatin sponge were performed in 12 patients (group D). The other 57 patients only received liver resection (non-TACE group). The extent of apoptosis was analyzed by transferase-mediated dUTP nick end labeling (TUNEL) staining. The expressions of Bcl-2 and Bax protein were detected by immunohistochemical method.

RESULTSThe apoptotic index(AI) and level of Bax protein in HCC cells were significantly higher in groups A, B, C and D than those in the non-TACE group (P < 0.05). The level of Bcl-2 protein and ratio of Bcl-2 to Bax protein of HCC cells were significantly lower in Groups A, B, C and D than those in the non-TACE group (P < 0.05).

CONCLUSIONPreoperative TACE regimens may enhance apoptosis of HCC cells by up-regulating the expression of Bax protein and down-regulating the expression of Bcl-2 protein and ratio of Bcl-2 to Bax protein expression.

Adult ; Aged ; Apoptosis ; Carcinoma, Hepatocellular ; chemistry ; pathology ; therapy ; Chemoembolization, Therapeutic ; Ethanol ; administration & dosage ; Female ; Humans ; Iodized Oil ; administration & dosage ; Liver Neoplasms ; chemistry ; pathology ; therapy ; Male ; Middle Aged ; Proto-Oncogene Proteins ; analysis ; Proto-Oncogene Proteins c-bcl-2 ; analysis ; bcl-2-Associated X Protein

Objective To study the effect of high-flow oxygen insuffiation (HFOI) via nasal cannula in reduction in re-intubation rate,length of ICU stay and improvement of respiratory function in patients at high risk of re-intubation after weaning from mechanical ventilation assessed by lung ultrasound score (LUS).Methods Single center randomized(random number) clinical trial was carried out in one intensive care units in China from May 2016 to May 2017 including critically ill patients ready for planned extubation with high-risk factors for re-intubation assessed by LUS when the LUS ≥ 14 was considered to be high risk.The comparisons of the length of ICU stay,re-intubation rate in case of respiratory failure,respiratory rate pulse rate SaO2 PaO2/FiO2 of patients at 6 h,24 h and 48 hours after extubation were made between HFOI and conventional oxygen therapy (COT) group.Results During the study period,32 patients were enrolled in the study.Of them,15 were assigned in HFOI group and 17 in COT group.The length of ICU stay (8.0±2.4)days vs.(10.9±3.5) days and re-intubation rate (6.7％ vs.23.5％) were significantly different between two groups (P＜0.05).The respiratory rate pulse rate SaO2 and PaO2/FiO2 of patients at 6 h after ex-tubation in HFOI group were improved than those in COT group (P＜0.05);and the SaO2 and PaO2/FiO2 of patients 24 h and 48 h after ex-tubation in HFOI group had much more improvement than those in COT group (P＜0.05).Conclusion Among high-risk adults who assessed by lung ultrasound score,high-flow oxygen therapy could reduce re-intubation rate length of ICU stay and improve the respiratory function.High-flow oxygen therapy may offer advantages for these patients.

OBJECTIVETo observe the fibrosis of skin after damage to the fat dome structure in skin of pig.

METHODSTotally 4 pieces of skin grafts of intermediate thickness in the size of 5 cm × 5 cm were obtained from both sides beside the spine of back in each of the 4 female red Duroc pigs with pedicle on one side with Humby knife performed by burn specialists, who were rich in clinical experience. These skin grafts were assigned as thin dermis group (TD). Pedicled tissue grafts in the size of 5 cm × 5 cm with the thickness of 1.5 mm were obtained within the wounds resulted from former incision with the same method mentioned above, and these tissue grafts were set as fat dome group (FD). The above-mentioned two groups of skin grafts were sutured back in situ immediately after completion of the former procedures. On post surgery day (PSD) 7, 14, and 21, 5 wounds were respectively selected according to the random number table for gross observation of the surgical areas. Tissue samples were obtained from corresponding surgical area deep to the deep fascia after gross observation at above-mentioned time points. Some of the tissue samples were used for observation of distribution of collagen fibers in the regions of operation of both groups of skin grafts with HE staining, and the breadth of fibrosis was measured; some of the tissue samples were used for observation of distribution of type I or III collagen fibers in the regions of incision of both two groups of skin grafts with Sirius red staining. Data were processed with two independent sample t test.

RESULTSA little scab on the edge of wounds was observed on PSD 7; all the wounds were healed on PSD 14; a few hairs were observed growing in the surgical area on PSD 21. HE staining showed that traces of incision were observed in the superficial layer of dermis and at the junction between dermis and fat dome at each time point; profuse hyperplasia of collagen fibers with parallel and orderly arrangement were observed in the region of incision of skin grafts in groups TD and FD at each time point. The breadth of fibrosis of the region of incision of skin grafts was respectively (251 ± 31), (240 ± 3 7), and (342 ± 69) µm in group TD, (239 ± 36), (286 ± 61), and (332 ± 28) µm in group FD on PSD 7, 14, 21, without significantly statistical difference (with t values respectively 0.750, -1.971, and 0.375, P values above 0.05). Sirius red staining showed that large amount of type III collagen fibers and small amount of type I collagen fibers arranging parallelly were present in the region of incision of skin grafts in groups TD and FD at each time point.

CONCLUSIONSUnder the circumstances of relatively intact restoration of dermal tissue, no excessive fibrosis was observed after simple incisional injury of fat dome in skin of pig.

Animals ; Burns ; surgery ; Dermis ; surgery ; transplantation ; Female ; Fibrosis ; complications ; Graft Survival ; Male ; Skin ; Skin Transplantation ; methods ; Skin, Artificial ; Swine ; Wound Healing