Objective To investigate the effects of preemptive analgesia with nalbuphine on inflammatory factors in elderly patients undergoing thoracotomy. Methods Eighty elderly patients with ASA I or Ⅱ undergoing thoracotomy were randomly allocated into control group (group C:n=40) and Nalbuphine group (group N:n=40). Each patient received 20 mg of Nalbuphine intravenously before induction of anesthesia in group N and same volume of normal saline in group C. Plasma levels of IL-10, TNF-α and IL-6 were measured at different time points. The VAS was recorded and the MAP, HR, SpO2 and PaCO2 were monitored. Results Compared with group C, the levels of TNF-α and IL-6 were lower, while IL-10 was higher at T2, T3, T4 in group N(P < 0.05). The VAS, MAP, HR and PaCO2 in group N were lower than those in group C at T2, T3 and T4 (P < 0.05). Conclusion Preemptive analgesia with Nalbuphine can improve the efficacy of postoperative analgesia and effectively reduce inflammatory reaction in the old patients undergoing thoracotomy.

BACKGROUND:Currently, the enzymatic digestion combined with magnetic activated cel sorting for isolating microvascular endothelial cel s are cumbersome and do harm to cel s. Therefore, how to simplify the isolation and culture of human dermal microvascular endothelial cel s to obtain highly purified endothelial cel s in vitro becomes a hotspot.
OBJECTIVE:To explore a simple and effective cultivation method of microvascular endothelial cel s from diabetic patient skins in vitro, and to detect the cel growth.
METHODS:Diabetic patients with chronic foot wounds after amputation were enrol ed to col ect the limb proximal skin and topical skin around the wound superficial dermal tissue. Human dermal microvascular endothelial cel s were obtained using adherent method and trypsin method, fol oewd by purified utilizing trypsin digestion and repeated attachment method when passage culture.
RESULTS AND CONCLUSION:Human dermal microvascular endothelial cells were obtained successfully, Primary cultured endothelial cells completely adhered to the wall at 24 hours, entered the logarithmic phase at the 10th day, and the cell concentration reached 80%at the 12th-13th day. While the passage cells grew more actively than primary cells, and fully covered the bottom in a“cobblestone”arrangement after 5-7 days of culture. Immunohistochemical staining showed that cultured cells were positive for FVIII and CD31-associated antigens with 100%positive rate. MTT assay showed that cell growth curves of 2, 4, and 5 generations of dermal microvascular endothelial presented the invertedSshape. These results suggest that abundant highly purified human dermal microvascular endothelial cells can be obtained through the adherent method and a small amount of short-term trypsin method.

Objective To measure the organ weights , blood physiological and biochemical parameters of KK /Upj-Ay/J.Methods KK/Upj-Ay/J mice at five and ten weeks of age were selected , and the organ weights , blood physiological and biochemical parameters were observed .Results Parts of the organ weights , blood physiological and biochemical parameters of different ages and sexes were significant differences .The fasted blood glucose of KK/Upj-Ay/J mice reached 7.0mmol/L at 10 weeks of age .Conclusion The results show that the organ weights , blood physiological and biochemical parameters are affected by age and gender of KK /Upj-Ay/J mice.The fasted blood glucose reached the diabetes level at 10 weeks of age .

Objective To observe the influence of Huiyang Shengji Ointment and its modified formulae on interleukin-1 (IL-1α) and thromboxane B2 (TXB2) in diabetic rats with chronic skin ulcers, and explore the mechanism for promoting the healing of ulcer.Methods Six out of 160 rats were randomly selected as a blank group, without any further processing. The remaining rats were made diabetic model and randomly divided into five groups after 2 weeks:1 d, 3 d, 5 d, 7 d and 14 d groups. Then, these groups were further divided into normal group (Vaseline ointments), model group (Vaseline ointments), Huiyang Shengji Ointment group (whole formula Ointment), Wenyang Yiqi group (Yiqi group, modified Wenyang Yiqi formula ointments) and Huoxue Shengji group (Huoxue group, modified Huoxue Shengji formula ointments). Normal group and model group were given Vaseline ointments;whole formula group, Yiqi group and Huoxue group were given corresponding ointment. Normal group used the method of skin excision, while other groups used STZ injection-hydrocortisone interference-skin excision-foreign body embedded preparation of composite factors for chronic skin ulcer model. After the appropriate treatment period, the rats were executed and tested for the contents of IL-1α and TXB2 in serum by enzyme-linked immunosorbent assay of five time points.Results In treatment 3 d, the contents of IL-1α in Yiqi group were significantly higher than the blank group, model group, whole formula group and Huoxue group (P<0.05). In treatment 5 d, the contents of IL-1α in whole formula group were significantly higher than the blank group and model group (P<0.05). In treatment 7 d, the contents of IL-1α in each treatment group were significantly higher than blank group and model group (P<0.05), and the whole formula group was higher than the Yiqi group and Huoxue group. In treatment 14 d, the contents of IL-1α in model group and Huoxue group were lower than the blank group (P<0.05). In treatment 3 d, the contents of TXB2 in normal group and the whole formula group were higher than the blank group (P<0.05). In treatment 5 d, the contents of TXB2 in whole formula group were higher than the blank group and the normal group (P<0.05). In treatment 7 d, the contents of TXB2 in Yiqi group were higher than the blank, the model, the whole formula and Huoxue groups (P<0.05). In treatment 14 d, the contents of TXB2 in Huoxue group were higher than the blank and model group (P<0.05), and the contents of TXB2 in the blank group and normal group was lower than those treatment groups (P<0.05).Conclusion Huiyang Shengji Ointment and its modified formulae could promote inflammation, stimulate secretion of inflammatory cytokines, while Yiqi Wenyang ointments played a more active role in promoting inflammation of the early phase of wound surface.

Objective To observe and compare the clinical efficacies between electroacupuncture and warm needling in treating low back pain.Method Seventy-eight eligible low back pain patients were randomized into group A of 28 cases, group B of 26 cases, and group C of 24 cases. Group A was intervened by electroacupuncture, group B was by warm needling, and group C was by medication. The short-form McGill Pain Questionnaire, Japanese Orthopaedic Association Scores (JOA), and Oswestry Disability Index were observed before and after treatment, and the therapeutic efficacies were compared.Result In group A, the McGill item scores [Sensory Pain Rating Index (S-PRI), Affective Pain Rating Index (A-PRI)] respectively after 1-week and 2-week treatment as well as in the 1-month and 3-month follow-up were significantly different from that before treatment (P<0.01,P<0.05). In group B and C, the McGill item scores after 2-week treatment and in the 1-month and 3-month follow-up were significantly different from that before treatment in the same group (P<0.01,P<0.05). The JOA and Oswestry scores were significantly changed respectively after 1-week and 2-week treatment and in the 1-month and 3-month follow-up in the three groups compared with that before treatment (P<0.05,P<0.01). After 1-week and 2-week treatment and in the 1-month and 3-month follow-up, the JOA and Oswestry scores in group A were significantly different from that in group C (P<0.05,P<0.01). In the 1-month and 3-month follow-up, the JOA scores in group B were significantly different from that in group C (P<0.05). The total effective rate was 85.7% in group A and 73.1% in group B, both significantly higher than 58.3% in group C (P<0.05). Conclusion Electroacupuncture and warm needling both can produce a significant efficacy in treating low back pain, but warm needling acts comparatively slowly and is less safe.

Objective To explore the effect of total saponins from Medicago polymorpha (TSMP) on glucose metabolism in type 2 diabetes mellitus (T2DM) rats and its possible mechanism.Methods T2DM rats were established by feeding with fat diets and intraperitoneally injecting with STZ 30 mg/kg.The rats were divided into control group,model group,metformin group (0.2 g/kg),TSMP high dose group (1.4 g/kg) and TSMP low dose group (0.7 g/kg),which were administrated for four weeks.At the end of administration,blood samples were collected to determine insulin resistance index (IRI),levels of fasting blood glucose (FBG),glycosylated hemoglobin (HbAlc),hepatic glycogen,interleukin (IL)-1 β,tumor necrosis factor (TNF)-α,free fatty acids (FFA) and Leptin,and activities of pyruvate kinase (PK),hexokinase (HK),glucose-6-phosphatase (G-6-Pase),fructose-1,6-bisphosphatase and glucokinase (GK).Western blot was used to detect the expression levels of G-6-Pase and phosphoenolpyruvate carboxykinase (PEPCK) proteins.Results Compared withthe model group,high dose (1.4 g/kg) TSMP decreased IRI,levels of FBG,HbA1 c,IL-1β,TNF-α,FFA and Leptin,and activities of G-6-Pase and fructose-1,6-bisphosphatase in T2DM rats (P＜ 0.05),while increased level of hepatic glycogen,and activities of PK,HK and GK (P＜0.05).Moreover,high dose (1.4 g/kg) TSMP down-regulated expression levels of G-6-Pase and PEPCK protein in liver tissues.Conclusion Chronic administration of TSMP can improved glucose homeostasis in T2DM rats,which might be related to promoting utilization of glucose,and alleviating inflammatory and insulin resistance.

Objective: To investigate the value of short tandem repeat (STR) genotyping in the diagnostic workup of molar and non-molar gestations with correlation of histological characteristics. Methods: Six hundred and fifty-six cases were selected based on clinically suspected hydropic abortion and/or molar pregnancy from July 2015 to September 2017 at Beijing Obstetrics and Gynecology Hospital. DNA was extracted from dissected chorionic villi and paired maternal endometrial FFPE tissue samples by Simplex OUP™ FFPE DNA Tissue Kit. STR genotyping was performed by PowerPlex 16 HS system. Results: DNA genotyping was informative in 649 of 656 cases, leading to identification of 215 hydatidiform mole gestations and 434 non-molar gestations. Most of non-molar gestations (375 cases, 86.4%) were diploid hydropic abortion. Various trisomy syndromes were found (53 cases, 12.2%), including trisomy 2, 3, 4, 7, 8, 13, 16 and 21. Only 2(0.5%) digynic triploid gestations were detected. Moreover, 4 cases (0.9%) of uniparental disomies (homologous or heterologous) were found. There were 196 cases with histologic diagnostic suspicious of hydatidiform moles were accurate sub-classified. Among them, 59 cases hydatidiform moles were under-diagnosed as diploid hydropic abortions, and 28 cases diploid hydropic abortions were over-diagnosed as hydatidiform moles.Compared with partial moles(PHM), there were no specific histomorphological features between the various types of non-molar gestations and partial moles for definitive diagnostic separation. There was no significant difference in the expression of p57^kip2 among PHM, trisomy and diploid hydropic abortions group (P=0.247). Conclusions STR genotyping can distinguish non-molar gestations from early hydatidiform moles, and efficiently avoid misdiagnosis based only on histological evaluation. Therefore, using STR genotyping, not only can the overdiagnosis of non-molar pregnancy be avoided, but also individualized management can be offered to patients including monitoring of serum hCG.

Objective: To investigate the frequency of KRAS mutation in mucinous epithelial lesions of the endometrium, and analyze the correlation between KRAS mutation and the clinicopathologic features. Methods: The cohort included forty-three cases of mucinous epithelial lesions of the endometrium selected from July 2015 to October 2017 from Beijing Obstetrics and Gynecology Hospital, and 22 control cases. Genomic DNA was extracted from formalin-fixed paraffin-embedded tissue sections. Polymerase chain reaction amplification for KRAS exons 2 and 3 was performed, followed by sequencing using capillary electrophoresis. The Fisher exact test was used to compare the prevalence of KRAS mutation among the different groups. Results: The patients′age ranged from 33 to 77 years [mean (55.12±9.34) years, median 55 years]. None of the eight cases of endometrial hyperplasia with mucinous differentiation without atypia showed KRAS mutation. The frequency of KRAS mutations was 1/10 in endometrial atypical hyperplasia, 1/12 in endometrioid carcinoma, 4/11 in endometrial atypical hyperplasia with mucinous differentiation (EAHMD), 6/15 in endometrioid carcinoma with mucinous differentiation (ECMD) and 8/9 in mucinous carcinoma (MC), respectively. The differences were statistically significant between MC versus EC (P<0.01) and MC versus ECMD (P<0.05). Conclusion The high frequency of KRAS mutation in EAHMD, ECMD and MC indicates that KRAS mutational activation is implicated in the pathogenesis of endometrial mucinous carcinoma.

OBJECTIVE To systematically evaluate the efficacy and safety of the four most common cell therapies， namely purified CD34+ （PCCs）， bone marrow mononuclear cells （BMMNCs）， bone marrow mesenchymal stem cells （BMMSCs） and peripheral blood mononuclear cells （PBMNCs） in the treatment of critical limb ischemia （CLI）. METHODS PubMed， Scopus， Embase， Cochrane Central Register of Controlled Trials （CENTRAL） and Web of Science databases were searched from the establishment of each database to June 2023 to collect randomized controlled trials （RCTs） comparing the efficacy and safety of four different cell therapies， namely PCCs， BMMNCs， BMMSCs and PBMNCs， with other cell therapies or standard therapy （ST） in the treatment of CLI. The outcomes indexes included amputation rate， ankle-brachial index （ABI）， transcutaneous oxygen partial pressure （TCPO2）， ulcer healing rate， pain-free walking distance （PFWD） and angiogenesis. After data extraction from clinical studies that met the inclusion criteria， the RoB 2.0 tool was used to assess the risk of bias， and Stata 15.0 software was used for statistical analysis. RESULTS Meta-analysis included 22 studies， involving 1 318 patients. The treatment groups involved 4 types of cell therapies， namely PCCs，BMMNCs， BMMSCs， and PBMNCs. Network meta-analysis showed that the amputation rates of the four cell therapies groups were lower than that of ST group， and only the difference in PBMNCs group was statistically significant（P＜0.05）. Four cell interventions were better than ST in improving ABI （P＜0.05）， and BMMNCs had the most significant effect on improving ABI. PBMNCs and BMMNCs groups had statistically significant differences in improving TCPO2， compared with ST group and BMMSCs group （P＜0.05）. Four cell interventions were better than ST in improving ulcer healing rate， among which BMMNCs group had no statistical difference with ST group （P＞0.05）； ulcer healing rates of the other three groups were higher than that of ST group （P＜0.05）， and those of PBMNCs and BMMSCs groups were significantly higher than that of BMMNCs group （P＜ 0.05）. BMMSCs group had a significantly better effect on improving the PFWD of patients than the ST group after transplantation， with statistical significance （P＜0.05）， but there was no significant difference in PBMNCs and BMMNCs groups compared with ST group （P＞0.05）. The three cell therapies of BMMSCs， BMMNCs and PBMNCs had a significantly better effect on angiogenesis than the ST group， and the BMMSCs group had a significantly better effect than the BMMNCs and PBMNCs groups， with statistical significance （P＜0.05）. CONCLUSIONS The four cell therapies can improve the prognosis of CLI patients to varying degrees. PBMNCs show the lowest amputation rate after transplantation and have the most significant effect on improving TCPO2 and improving the ulcer healing rate. BMMNCs possess the most significant effect on improving ABI. BMMSCs represent obvious advantages in PFWD and angiogenesis.

【Objective】 To study the serological characteristics and genetic background of 20 samples with ABO blood group discrepancies in Shenyang. 【Methods】 Serological test, polymerase chain reaction-sequence specific primer (PCR-SSP) and sequencing of the full coding of ABO gene and the Intron 1 were conducted in 20 samples with ABO blood group discrepancies. 【Results】 Ten subtypes (Am, Bw, Bx, B3,, A2B, AxB, A2Bw, A2Bx, AwB and ABw) were detected in 20 samples, with AB subtype as the dominant. Sixteen ABO alleles were found, including 5 common alleles (A1.01, A1.02, B. 01, O. 01.01 and O. 01.02), nine rare alleles (AW.37, BW.03, BW.08, B3.07, cisAB.02, cisAB.03, cisAB.06, BA.04 and O. 01.04) and two novel alleles (AM.03 and cisAB.07). The AM.03 allele had a nucleotide change at position 912 (C to A) compared with A1.02 allele, which resulted in an amino acid substitution (S304R). The cisAB.07 allele was observed a missense mutation at position 797 (T to C) which resulted in an amino acid substitution (M266R) compared with B. 01 allele. The serologic had been changed, and both A antigen and B antigen were expressed. 【Conclusion】 The study revealed the genetic background of 20 samples with ABO blood group discrepancies, and two new alleles as ABO*AM.03 (912 C to A) and ABO*cisAB.07 (797 T to C) were first reported.