OBJECTIVE:To establish a method for determining recombinant human calmodulin B subunit(rhCNB)in rat plas-ma,and study its pharmacokinetics characteristics. METHODS:ELISA double-antibody sandwich method was adopted. 1 μg/ml rhCNB monoclonal antibody mAb was wrapped,added to the to-be-test sample,rhCNB polyclonal antibody pAb(dilution ratio of 1∶5 000)and HRP-labeled conjugate of anti-IgG(dilution ratio of 1∶10 000)were added. Using tetramethylbenzidine for develop-ing,microplate reader was conducted in wavelength of 450 nm to determine the absorbance value(OD value)and plasma concen-tration of 6 rats after 2,15,30,60,120,240,480,720 min of iv 2.5 mg/kg rhCNB,and the pharmacokinetic parameters were calculated by BAPP 3.0 software. RESULTS:The linear range of rhCNB were 0.195-12.5 ng/ml(r2=0.995 0),lower limit of quan-titation was 0.195 ng/ml,accuracy were 97.300%-103.622%(RSD<7.5%,n=6);RSDs of within-batch,inter-batch,freezing and thawing 3 times were no higher than 8.5%(n=6,18,15). rhCNB pharmacokinetics characteristics in rat fitted to two-com-partment model,AUC0-720 min was 173.038 mg·min/L and t1/2 was 94.62 min. CONCLUSIONS:The established method has high specificity and sensitivity,good accuracy and precision,which can be used for rhCNB quantitative detection and pharmacokinetics study in biological samples.

Objective To explore an anatomical basis for the lateral tarsal artery pedicle flap on front and lateral compartment of leg and the feasibility of repairing skin defects on forepart of feet. Methods The branches, course and anastomosis of the lateral tarsal artery, perforator of peroneal artery up external malleolus, superficial peroneal artery were studied in 20 legs of adult cadavers.The flap was designed on these grounds. 8 cases repaired by lateral tarsal artery pedicle flap on front and lateral compartment of leg, 5 cases of skin defects on dorsum of foots, 3 cases of skin defects on footplates.The area of defect on forepart of foot was 5 cm× 4 cm-cm × 5 cm. The donor sites were resurfaced with skin grafts or sutured directly. The lateral tarsal artery, perforator of peroneal artery up external malleolus, perforator of anterior tibial artery superficial peroneal artery were anastomosed each other, formed single band blood vessel axle on lateral foot, fore external malleolus, front and lateral compartment of leg. The area of flap was 6 cm × 4 cm - 10 cm × 6 cm.Results All of the flaps survived completely. All cases were followed up, followed up 6- 12 months, averaged 8 months. The color, appearance and texture of the flaps were good, without ulcer on the flap. The patients can walk freely. Conclusion The flap on front and lateral compartment of leg should be designed according to the lateral tarsal artery. Blood supply of flap was reliable, little trauma. The flap's vessel pedicle is enough long. It could repair any defect on forepart of foots.

OBJECTIVETo investigate the characteristics of the hepatic pathological and clinical features of patients with hepatitis B virus (HBV) in immune tolerant stage and find the better measure of diagnosing patients chronic infected by HBV in immune tolerant phase.

METHODS135 patients with HBV chronic infection and persistently normal serum alanine aminotransferase (ALT) levels were involved in this study, whose serum HBeAg and HBV DNA were positive. Statistical analysis included the ages, sex, serum levels of HBVDNA, ALT and histological grade. The grades of inflammation and fibrosis were obtained through hepatic biopsy performed on all the patients.

RESULTSMean age in those patients was 22.61 +/- 8.95 years old. All those patients were divided into two groups according to histological grade: low- histological grade group, G < or = 1 and S < or = 1; and high-histological grade group, G > or = 2, S > or = to 2. Levels of histological grade were low in most of patients (99/135). Patients of low-histological grade had no difference in age, sex and family history of HBsAg carriers. Compared with low-normal ALT (ALT less than 30U/L), those with high-normal ALT (ALT > or = 30U/L) had a greater frequencies of high-histological grade. Compared with high HBVDNA (HBVDNA > or = 10(7) copies/ml), those with low HBVDNA (HBVDNA less than 10(7) copies/ml) had a greater frequencies of high-histological grade.

CONCLUSIONLevels of histological grade were low in most of patients with HBV chronic infection, serum HBeAg and HBVDNA positive, persistently normal serum ALT levels, but some of them were high-histological grade. It showed those patients were not all in immune tolerant stage of chronic HBV infection. Examination of ALT and HBVDNA are helpful to evaluate hepatic pathological damage for them.

Adolescent ; Adult ; Alanine Transaminase ; blood ; Child ; Child, Preschool ; DNA, Viral ; blood ; Female ; Hepatitis B, Chronic ; immunology ; pathology ; virology ; Humans ; Immune Tolerance ; Male ; Middle Aged

OBJECTIVETo provide content-measuring evidences for establishing the quality criteria of dexamethasone acetate ulcer pasta.

METHODSContent of dexamethasone acetate was assayed by the isonicotinyl hydrazine (INH) colorimetry.

RESULTSThe good linear range was 6-16 microg/mL (r = 0.999 1). The average recovery of dexamethasone acetate was 98.9%, and relative standard deviation was low than 1% (n = 5).

CONCLUSIONIt is easy, convenient, accurate and dependable for INH colorimetry to determine the content of dexamethasone acetate.

Chromatography, High Pressure Liquid ; Dexamethasone ; analogs & derivatives ; Humans ; Oral Medicine ; Oral Ulcer

HBV-associated acute-on-chronic liver failure is prevalent in mainland China.The prognosis of HBV-ACLF is poor.The mortality of HBV-ACLF is approximately 80％.Therefore,a prognostic indicator was needed in order to allow us to intervene as soon as possible.The model for end-stage liver disease (MELD) scoring system is widely used to predict the prognosis of liver failure.However,the assessment is too complex to restrict its application.This study aimed to investigate the expression ofIP-10 in peripheral blood mononuclear cells (PBMC),in order to explore the relationship between the expression and prognosis of patients with HBV-ACLF.The mRNA level of IP-10 in PBMCs were analyzed in 80 patients with HBV-ACLF,40 patients with chronic hepatitis B (CHB)and 40 healthy people by fluorescent quantitative PCR.IP-10 mRNA level was significantly higher in the HBV-ACLF group than in the other two groups (P＜0.01).Group with MELD score below 30 had lower IP-10 mRNA level than group with MELD score over 30 (P＜0.05).The IP-10 mRNA level in PBMCs in positive group was higher than that in negative group (P＜0.01).With a threshold of 0.925,the area under the receiver operating characteristic (ROC) curves was 0.815.These findings suggest that assessment of IP-10 mRNA level in the PBMCs would be helpful for evaluating the disease severity and prognosis in patients with HBV-ACLF.

In order to compare the differences of 35 Menthae Herba samples collected on the market and at producing areas, the contents of six total terpenoids, the essential oil and chromatographic fingerprints were analyzed, which provided evidences for drawing up the commodity specifications and grading criteria of Menthae Herba. GC-MS method was used to analyze the chemical constituents of 35 different samples. The chromatographic fingerprints obtained by using GC were then evaluated by similarity analysis, hierarchical clustering analysis and principal component analysis. The relativity between the content of six terpenoids and the essential oil were studied. In this study, the chemical profiles of 35 samples from different producing areas had significant disparity. All samples collected in the report could be categorized into four chemical types, L-menthol, pulegone, carvone and L-menthone, but the chemical profiles had no relationship with the areas. The chromatographic fingerprints of the samples from different types were dissimilar, while the different producing areas were difficult to be separated. It was indicated that the content of volatile oil was positively correlated with the content of L-menthol and the sum of six total terpenoids. The content of the essential oil, L-menthol and the sum of six total terpenoids of Menthae Herba were considered as one of the commercial specifications and grading criteria. These results in the research could be helpful to draw up the commercial specification and grading criteria of Menthae Herba from a view of chemical information.
Cluster Analysis ; Gas Chromatography-Mass Spectrometry ; Mentha ; chemistry ; Oils, Volatile ; analysis ; Principal Component Analysis ; Terpenes ; analysis

Objective:To evaluate the predictive value of stair climbing test combined with arterial blood gas analysis on postoperative complications in lung cancer patients with limited pulmonary function.Methods:A total of 1 231 hospitalized lung cancer patients with limited pulmonary function dating from August 2012 to August 2020 were retrospectively reviewed. Included in the cohort were 766 of patients who underwent stair climbing test(SCT) preoperatively and completed data collection. Patients were grouped according to their general condition, past medical history, surgical approach, pulmonary function test(PFT) and SCT results. Comparison of the postoperative cardiopulmonary complication rates were made between different groups, and independent risk factors were identified.Results:A total of 182 cardiopulmonary-related complications occurred in 144 cases, accounting for 18.8% of the entire cohort. Perioperative mortality rate was 0.9%(7/766). The rate of postoperative cardiopulmonary complications was significantly different between the groups stratified by gender, age, smoking index, PFT index(FEV1%, DLCO%), SCT results(height achieved, speed, changes in heart rate and oxygen saturation of the arteries before and after the test), ASA score, surgical approach(VATS/Open), resection range(Lobectomy/Sublobectomy), anesthetic duration, blood loss volume, etc. Logistic regression analysis showed that only height achieved( P<0.001), changes in heart rate( P<0.001), changes in oxygen saturation of the arteries( P=0.001), resection range( P=0.006) and anesthetic duration( P=0.025) were independent risk factors for cardiopulmonary-related complications in lung cancer patients with limited pulmonary function. Conclusion:The stair climbing test combined with arterial blood gas analysis could be used as a preoperative screening method for lung cancer patients with limited lung function and may have a predictive value for postoperative cardiopulmonary-related complications.

BACKGROUNDTo investigate the prognostic significance and role of coagulation factor V (CFV) levels in clinical diagnostic criteria for severe hepatitis.

METHODSThe CFV level and prothrombin activity (PTA) were tested by turbidimetry for 129 times in 58 patients with severe hepatitis. Comparative studies and clinical significance of CFV and PTA were analyzed by SPSS and SDAS softwares.

RESULTS1. The levels of CFV and PTA were 15.3%+/-9.7% and 23.5%+/-10.0%, respectively, at the onset of severe hepatitis. 2. The mortality of severe hepatitis gradually increased with the gradual decrease of CFV or PTA during the most severe stage of the illness (P=0.000). 3. The levels of CFV and PTA decreased continually and rapidly in patients who died but gradually increased in survivors. The decrease or increase of PTA preceded that of CFV on the exacerbation or convalescent stage. 4. Hepatic encephalopathy occurred in 14 cases (24.14%). In 10 cases, it occurred in the terminal stage of the illness, far later than the time of the decrease of CFV. 5. The level of CFV was closely related to PTA (the correlation coefficient was 0.812), the level of CFV was almost consistent with that of PTA.

CONCLUSION1. The level of CFV is an important prognostic indicator in severe hepatitis and is more specific than PTA. 2. Simultaneous determination of CFV and PTA may be helpful in earlier and more accurate diagnosis of severe hepatitis. 3. Possible use of CFV as one of the criteria for liver transplantation in patients with severe hepatitis should be studied.

Adult ; Aged ; Diagnostic Techniques and Procedures ; Factor V ; analysis ; metabolism ; Female ; Hepatitis ; diagnosis ; metabolism ; Humans ; Male ; Middle Aged ; Nephelometry and Turbidimetry ; methods ; Prognosis ; Prothrombin ; analysis ; metabolism ; Young Adult

OBJECTIVETo discuss the application of medial planta island flaps pedicled with anterior tibial artery perforator in front of inner malleolus for repairing small wounds around ankle.

METHODSFrom Jan. 2005 to Jun. 2009, 10 cases with small wounds around ankle were treated with medial planta island flaps pedicled with anterior tibial artery perforator. The flap size ranged from 7.5 cm x 2.8 cm to 13.0 cm x 5.0 cm. The wounds at the donor sites were covered with skin grafts.

RESULTSAll the 10 flaps and skin grafts were survived with primary healing. The patients were followed up for 6-12 months with satisfactory cosmetic results. The 2-point discrimination was 4-6 mm when the proximal end of saphenous nerve was not injured, and it was 9-10 mm when the nerve was injured or cut off. The patients could walk with no occurrence of ulcer in flaps or donor site.

CONCLUSIONSThe medial planta island flaps pedicled with anterior tibial artery perforator can effectively repair the small wounds around ankle with reliable blood supply.

Adult ; Ankle Injuries ; surgery ; Feasibility Studies ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Skin Transplantation ; Surgical Flaps ; blood supply ; Tibial Arteries ; surgery ; Treatment Outcome ; Young Adult

Hematopoietic stem cells (HSC) shift from fetal liver and spleen to bone marrow at neonatal stages and this movement may be due to inductive signals from different microenvironments. Mesenchymal stem cells (MSC) are the precursors of stromal cells in bone marrow microenvironments such as osteoblasts and endothelial cells. Some researchers speculated that fetal bone marrow before birth might be not perfectly suit HSC growth. However, it is still lack of direct evidence to prove this hypothesis. This study was aimed to compare the hematopoietic supportive capacity between human fetal and adult bone marrow MSC in vitro. Adult bone marrow MSC (ABM-MSC) were isolated from three healthy donors and fetal bone marrow MSC (FBM-MSC) were isolated from three fetuses between gestations of 19 to 20 weeks. After irradiation, MSC were co-cultured with CD34(+) cells isolated from umbilical cord blood in long-term culture-initiating cell (LTC-IC) assay. The colony number of colony forming cells (CFC) was counted and the phenotypic changes of co-cultured CD34(+) cells were analyzed by flow cytometry. Cytokine expressions in both kinds of MSC were detected by reverse transcription polymerase chain reaction (RT-PCR). The results showed that ABM-MSC had a stronger hematopoietic supportive capacity than FBM-MSC. Both of them enhanced the differentiation of CD34(+) cells into myeloid lineages. Cytokines were expressed differently in ABM-MSC and FBM-MSC. It is concluded that ABM-MSC possess more potential application in some treatments than FBM-MSC, especially in hematopoietic reconstitution.
Adult Stem Cells ; cytology ; Bone Marrow Cells ; cytology ; Cell Differentiation ; Cells, Cultured ; Fetus ; Flow Cytometry ; Hematopoietic Stem Cells ; cytology ; Humans ; Mesenchymal Stromal Cells ; cytology