Humanin(HN,its analogue [Gly14]-Humanin,HNG)was originally identified as an endogenous peptide that protects neuronal cells from apoptosis induced by various types of Alzheimer's disease-related insults.But the relative low content of this peptide in its natural sources limits its further characterization.An expression vector pET32a/HNG was corstructed and transformed it into E.coli BL21 trxB(DE3).HNG was expressed as a fusion protein in the soluble fraction and was purified by nickel affinity chromatography.Subsequently,the purified fusion protein was cleaved by enterokinase and was further purified by reverse-phase HPLC.A 23 mg recombinant HNG(rHNG)from 1 L bacterial culture was purified.The molecular weight of rHNG determined by ESI-MS was 2876.5 Da which was the expected size for correctly processed peptide.The N-terminal amino acid sequence of rHNG determined by Edman degradation method is identical to the theoretical sequence.Neuroprotective bioassay studies of rHNG exhibited its potential neuroprotective effect comparable to that of the natural HNG peptide.

OBJECTIVETo compare the differences in the characteristic medicinal cupping therapy between the traditional cupping device and the innovated cupping device.

METHODSFifty patients of neck and low back pain were selected. The self-comparison was adopted. The cupping therapy was applied to the acupoints located on the left or right side with the traditional cupping device and the innovated cupping device. The cupping sites were centered at bilateral Quyuan (SI 13) and Dachangshu (BL 25). The cups were retained for 10 min. The traditional cupping device was the glass with smooth border, 100mL. The innovated cupping device was the vacuum-sucking cup. The operative time, medicinal leakage, comfort and cupping marks were observed for the two different cupping devices.

RESULTSThe operative time with the innovated medicinal cupping device was shorter obviously compared with the traditional one at Quyuan (SI 13) and Dachangshu (BL 25, both P<0. 05). The comfort with the innovated medicinal cupping device was remarkably improved as compared with the traditional one at the two acupoints (both P<0. 05). The medicinal leakage was similar between the two different devices during the cupping operation (both P>0. 05). The cupping marks with the innovated medicinal cupping device were much deeper than those with the traditional one after cupping therapy.

CONCLUSIONThe innovated cupping device is more convenent and comfortable in operation during the characteristic medicinal cupping therapy.

Acupuncture Points ; Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Low Back Pain ; therapy ; Male ; Medicine, Chinese Traditional ; instrumentation ; methods ; Meridians ; Middle Aged ; Neck Pain ; therapy ; Young Adult

Objective To administer a lower dose according to different indoor temperatures to give play to the effect of characteristic decoction cup hot compress and be able to save the cost of medicine. Method Twenty subjects were allocated, according to different indoor temperatures, to four groups, 5 cases each. Each subject received 10 sessions of decoction cupping with different doses. The decoction in all the cups was 45℃ before cupping. The temperature of decoction in the cups was measured after cupping. The average value of temperatures of the same doses at the same indoor temperature was calculated after cupping. The graph of average temperature curve of decoction in the cups was produced according to the indoor temperature after cupping with different doses. Finally obtained were the lowest doses of decoction with hot compress effect in the cup required at different indoor temperatures. Result To produce the effect of characteristic decoction cup hot compress, the decoction in the cup must be more than 55 mL in a indoor temperature of 10℃~14℃, more than 45 mL in a indoor temperature of 15℃~19℃ and more than 25 mL in a indoor temperature of 20℃~24℃. Only 20 mL of decoction was needed in indoor temperatures above 25℃. Conclusion A dose of decoction is flexibly selected according to a seasonal change in indoor temperature. That can not only have the effect of hot compress but also save the dosage.

Neurotensin receptor-1 (NTR1), which can stimulate the intracellular cascade signal pathway, belongs to the large superfamily of G-protein coupled receptors. NTR1 is related to the occurrence and development of several kinds of diseases. In order to screen the inhibitors for the cancers associated with NTR1 protein, we established a CHO (Chinese hamster ovary) cell line in which human neurotensin receptor-1 was highly expressed. The method is to construct the recombinant plasmid which was lysed with the hNTR1 gene and transfect it into CHO cells. After selected with G418, the cell line was evaluated by Western blotting analysis and calcium flux assays. Through the calcium flux assays on FlexStation 3, we got the EC50 value of neurotensin peptide which is the natural NTR1 agonist, and the IC 50 value of SR48692 which is the known NTR1 antagonist. The established human CHO/NTR1 cell line can be used to study the profile of NTR1 biological activity and further screen of NTR1 antagonists and agonists.
Animals ; CHO Cells ; Calcium Signaling ; Cricetinae ; Cricetulus ; Humans ; Pyrazoles ; pharmacology ; Quinolines ; pharmacology ; Receptors, Neurotensin ; antagonists & inhibitors ; genetics ; metabolism ; Transfection

Humanin (HN, its analogue [Gly14]-Humanin, HNG) was originally identified as an endogenous peptide that protects neuronal cells from apoptosis induced by various types of Alzheimer's disease-related insults. But the relative low content of this peptide in its natural sources limits its further characterization. An expression vector pET32a/HNG was corstructed and transformed it into E. coli BL21 trxB (DE3). HNG was expressed as a fusion protein in the soluble fraction and was purified by nickel affinity chromatography. Subsequently, the purified fusion protein was cleaved by enterokinase and was further purified by reverse-phase HPLC. A 23 mg recombinant HNG (rHNG) from 1 L bacterial culture was purified. The molecular weight of rHNG determined by ESI-MS was 2876.5 Da which was the expected size for correctly processed peptide. The N-terminal amino acid sequence of rHNG determined by Edman degradation method is identical to the theoretical sequence. Neuroprotective bioassay studies of rHNG exhibited its potential neuroprotective effect comparable to that of the natural HNG peptide.

OBJECTIVETo evaluate the effectiveness and safety of the meridian three-combined therapy for treatment of ordinary psoriasis.

METHODSA multi-central, randomized and positive drug controlled trial was adopted, and 233 cases were divided into an observation group of 116 cases and a control group of 117 cases. The observation group was treated with thread embedding at points, blood-letting puncture on the back of ear and auricular point pressing (i.e. meridian three-combined therapy). For thread embedding, 3-4 local points such ear points as Fei (CO14), Gan (CO12), Pizhixia (AT4), Shenmen (TF4) , cephalic and symmetric points of severe parts of the limb skin were selected according to the skin lesion position, and the treatment was given once each two weeks. For ear point tapping and pressing, 3-5 points were selected in each session. And the control group was treated with oral administration of Di yin Tablets, 5 tablets each time, twice each day. After treatment of 6 weeks, the clinical therapeutic effects, the score of skin lesion area, the scores for skin lesion severity and safety were compared in the two groups.

RESULTSThe markedly effective rate was 57.8 % in the observation group and 51.3% in the control group with no significant difference between the two groups (P>0.05); after treatment the scores for both the skin lesion area and the skin lesion severity were significantly decreased in the two groups compared with those before treatment (P<0.01), and with a significant difference between the two groups (P>0.05). And the incidence rate of the adverse reaction in the observation group was significantly lower than that in the control group.

CONCLUSIONThe meridian three-combined therapy is effective and safe for treatment of ordinary psoriasis.

Acupuncture Points ; Acupuncture Therapy ; Adolescent ; Adult ; Aged ; Bloodletting ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Humans ; Male ; Meridians ; Middle Aged ; Psoriasis ; drug therapy ; therapy ; Young Adult

OBJECTIVE: To compare the outcomes of endoscopic combined ultrasound-guided access (EUGA) with the conventional ultrasound-guided access (UGA) to achieve percutaneous renal access in endoscopic combined intrarenal surgery (ECIRS). METHODS: A retrospective review of 53 patients undergoing ECIRS to treat upper urinary tract calculi between January 2017 and October 2019 was con-ducted. All of the cases were of complex upper urinary tract stones larger than 2 cm in diameter. The com-plex stone situations, such as multiple renal calyces calculi or staghorn calculi necessitated ECIRS. Under general anesthesia, the patients were placed in the galdakao-modified supine valdivia (GMSV) position, thus allowing both antegrade and retrograde accesss. The patients were divided to UGA and EUGA groups according to the protocol of achieving percutaneous renal access. In 28 cases, endoscopic combined ultrasound-guided accesss were obtained. Puncture and dilation were performed under direct flexible ureteroscopic visualization, while percutaneous renal access of 25 cases were performed with the conventional technique employing ultrasound guidance. Demographic and perioperative information, such as stone burden, presence of hydronephrosis and number of calyces involved was compared. Primary outcomes included total operative time, renal access time, repeat puncture, hemoglobin level, perioperative complications, and stone-free rate. RESULTS: No major intra-operative complication was recorded in all the 53 ECRIS. No significant difference was observed between the groups in age and gender. There was no significant difference in body mass index[BMI (29.21±3.14) kg/m² vs.(28.53±2.56) kg/m²], stone burden (37.68±6.89) mm vs. (35.53±6.52) mm, number of calyces involved 2.72±0.68 vs. 2.86±0.71, presence of hydronephrosis (56.0% vs. 46.4%), total operative time (93.0±12.2) min vs. (96.8±14.2) min, hemoglobin level reduction (6.56±2.16) g/L vs. 97.54±2.64) g/L, stone-free rate (92.0% vs. 92.8%), hospital stay (5.52±0.59) d vs. (5.64±0.62) d, perioperative complication rate (8.0% vs. 7.2%). Two patients in EUGA group experienced perioperative complications (one urinary tract infection and one hematuria) while two patients in UGA group experienced perioperative urinary tract infection. None in both groups received blood transfusion. The patients undergoing EUGA had shorter renal access time [(4.0±0.7) min vs. (6.8±2.6) min, P < 0.01] and less repeat puncture (0 vs. 4 cases, P < 0.05). CONCLUSION EUGA is an optimal technique to establish percutaneous renal access in ECIRS, which minimizes access time and repeated procedures.
Humans ; Kidney Calculi ; Nephrostomy, Percutaneous ; Retrospective Studies ; Treatment Outcome ; Ultrasonography, Interventional ; Ureteroscopy

OBJECTIVETo detect the character of surgical treatment of massive soft tissue sarcoma in the shoulder girdle and analyze the impact factor to the result.

METHODSSeven patients with massive soft tissue sarcoma in the shoulder girdle were treated in our department between 2005 and 2009. There were 4 males and 3 females. All the patients were referred to our hospital after local recurrence post-operatively. The mean age was 43.8 years old (range 14 - 75). The maximum diameter of the tumor varied from 10 to 16 centimeters. All the patients were performed surgery, wide margin in 4 cases and marginal margin in 3 cases. Five were performed tumor resection and reconstruction with latissimus dorsi muscle flap transfer and skin graft. One was reconstructed with advanced skin flap and skin graft. The other one was treated with skin graft. The diagnosis included 3 malignant fibrous histiocytomas, 1 low grade myxoid fibrosarcoma, 1 Primitive neuroectodermal tumor, 1 rhabdomyosarcoma, 1 dermatofibrosarcomas protuberans. The MSTS score system was used to evaluate the shoulder function.

RESULTSSeven patients were followed up with long time. The mean follow up was 29 months (range 10 to 46 months). Two patients suffered local recurrence and one died of pulmonary metastasis 6 months after the second surgery for local recurrence. One patient suffered pulmonary metastasis. The last four patients were disease-free at the end of follow-up. The function of shoulder girdle was satisfactory. The mean MSTS score was 28.

CONCLUSIONSSoft tissue sarcomas in the shoulder girdle are easy to be misdiagnosed and mistreated. Wide surgical margin was the key impact factor to the local recurrence of soft tissue sarcoma in the shoulder girdle. The surgical margin and invasion of the tumor are the key factor to the prognosis. The soft tissue defect after surgery is often reconstructed by muscle flap transfer or skin flap transfer. The latissimus dorsi muscle flap transfer is often used.

Adolescent ; Adult ; Aged ; Female ; Humans ; Male ; Middle Aged ; Prognosis ; Retrospective Studies ; Sarcoma ; diagnosis ; surgery ; Shoulder ; pathology ; Soft Tissue Neoplasms ; diagnosis ; surgery ; Treatment Outcome ; Young Adult

OBJECTIVETo investigate the detection limit of multicolor combinational probe coding real-time PCR (MCPC-PCR) in detection of Salmonella and Staphylococcus aureus suspended in the food samples, and to apply MCPC-PCR to detect the samples of food poisoning.

METHODSSeries concentration of bacterium suspension (10(1) - 10(9) CFU/ml) was prepared by using 22 simulated samples including fresh meat and cakes and then MCPC-PCR was applied to detect Salmonella and Staphylococcus aureus in 22 samples. Enrichment broth of 101 frozen samples and 5 early patients' anal swabs in food poisoning cases were detected after the DNA samples were extracted.

RESULTSThe limits of MCPC-PCR assay in detecting Salmonella and Staphylococcus aureus were about 10(2) copies/test; 101 frozen enrichment broth of samples in food poisoning cases were detected by MCPC-PCR assay, of 23 positive samples, 18 were confirmed by bacteriology techniques; 96 samples detected by MCPC-PCR and bacteriology techniques had the same results, and the coincidence rate was 95.05%. Anal swabs, collected from 5 of early patients in a food poisoning case gave a clue to be Vibrio parahaemolyticus by MCPC-PCR assay and then were perfectly consistent with bacteriology assay.

CONCLUSIONAs a method of high sensitivity and good specificity, MCPC-PCR assay can quickly and conveniently detect multiple pathogens existing in food samples, therefore we recommend it to be used in rapidly screening or simultaneous detection of food-borne diseases.

Bacteriological Techniques ; methods ; Food Contamination ; analysis ; Food Microbiology ; Molecular Probe Techniques ; Molecular Sequence Data ; Polymerase Chain Reaction ; methods ; Salmonella ; genetics ; isolation & purification ; Sensitivity and Specificity ; Staphylococcus aureus ; genetics ; isolation & purification

OBJECTIVESTo study the effect of Houxue Zhitong decoration on the expression of the mitochondria-initiated apoptosis pathway and transmembrane protein I an II of the epidural scar tissue.

METHODSA total of 60 New Zealand rabbits (weight: 2.5-3.0 kg) were randomly divided into four groups, sham operation group (D, n=15), control group (B, n=14), sodium hyaluronate group (C, n=15), Houxue Zhitong decoration group (D, n=15). Except for group A, 1.0 cm x 1.0 cm dura mater uncovered area laminectomy was performed at I (4) and I(5), covered with 0.5 ml sodium hyaluronate in group C, covered with same amount of saline in group B and D. First 2 weeks after operation, animals in group D were lavaged with 2.5 ml/kg Houxue Zhitong decoction by one a day for 14 days. Five rabbits of each group selected randomly were killed in the 2,4,8 weeks after laminectomy. The specimens were prepared for determination of the expression of Fas and FasL, at scar tissue by semiquantitative reserve transcription-polymerase chain reaction (RT-PCR). The degree of scar adhesion was evaluated according by Rydell method.

RESULTSThe adhesion area in group B was larger than of group C and D in the 4th and 8th week. However, the number of fibroblasts and inflammantory cells in group D was the least among the three groups in the 8th week. At 2, 4, 8 weeks after operation, as compared with group B the expression of Fas, FasL of group C and D were decreased (P < 0.05). Especially, at 2 weeks, as compared with group B the expression of this two cytokines of group D was significant decreased (P < 0.05), too. In group C and D the duramater adhesion was decreased (P < 0.05). The proliferation of fibroblast and fibroblastic function were inhibited (P < 0.05).

CONCLUSIONHuoxue Zhitong is able to down-regulated the expression of Fas, FasL, which inhibited the proliferation of fibroblast, the fibroblastic function and the synthesis of extracellular matrix in the epidural scar tissue. It is an effective way of reducing peridural scar formation and prevent the failed back surgery syndrome.

Animals ; Apoptosis ; drug effects ; Cicatrix ; genetics ; metabolism ; pathology ; Drugs, Chinese Herbal ; pharmacology ; Epidural Space ; pathology ; Fas Ligand Protein ; genetics ; metabolism ; Female ; Gene Expression Regulation ; drug effects ; Rabbits ; Signal Transduction ; drug effects ; fas Receptor ; genetics ; metabolism