Detection of Exogenous γ-Hydroxybutyric Acid in Rat Blood Exosomes / 法医学杂志

OBJECTIVES@#To find a method to distinguish exogenous gamma-hydroxybutyrate (GHB) from endogenous GHB by establishing ultra-high performance liquid chromatography-mass spectrometry (UPLC-MS) based on exosome for quantitative detection of GHB in the rat blood.@*METHODS@#Adult male SD rats were divided into 1 h, 5 h, 10 h administration group and control group. After 1 h, 5 h and 10 h of single precursor of GHB gamma-butyrolactone (GBL) intraperitoneal injection in administration groups, 5 mL blood was collected from the abdominal aorta. Meanwhile, the control group was given a same dose of normal saline, and 5 mL blood was collected at 1 h. Among the 5 mL blood, 0.5 mL was directly detected by HPLC-MS after pretreatment, and exosomes were extracted from the remaining blood by differential centrifugation and detected.@*RESULTS@#The concentration of GHB in the control group was (87.36±33.48) ng/mL, and the concentration with administration at 1 h, 5 h and 10 h was (110 400.00±1 766.35) ng/mL, (1 479.00±687.01) ng/mL and (133.60±12.17) ng/mL, respectively. The results of exosome detection showed that no peak GHB signal was detected in the control group and the 10 h administration group, and the concentrations of GHB at 1 h and 5 h administration groups were (91.47±33.44) ng/mL and (49.43±7.05) ng/mL, respectively.@*CONCLUSIONS@#GHB was detected in blood exosome by UPLC-MS, which indicated that exogenous GHB could be detected in plasma exosomes, while endogenous GHB could not be detected, suggesting that this method may be used as a basis to determine whether there is exogenous drug intake.

Advances in the research of application of clinical decision support system in fluid resuscitation following severe burn / 中华烧伤杂志

Although guidelines and formulas have been developed through clinical practice to define infusion rate and volume, over- and under-resuscitation are still common, followed by increasing morbidity and mortality. In order to establish an effective management for early fluid resuscitation, the clinical decision support system (CDSS) has been established. The CDSS, by utilizing information systems coupled with decision support technology, could provide recommendations for the amount of fluid to be infused based on measured biological response. The results showed that patients treated with CDSS had a significantly lower mortality, increased ventilator-free days, and ICU-free days as compared with those treated with traditional fluid management. This article reviews the concepts as well as the result of recent clinical studies of CDSS for burn patients.

Amino acids 395-416 in DNA binding domain of STAT4 is involved in IL-12-induced nuclear import of STAT4 / 生理学报

The purpose of the present study is to explore the mechanism of IL-12-induced nuclear import of signal transducer and activator of transcription 4 (STAT4). Assayed by analyses of homology alignment of STATs, amino acids 395-416 in DNA binding domain was found to be a potential dimer-specific nuclear localization signal (dsNLS) of STAT4. Therefore, several plasmids were constructed. Wild-type STAT4 was inserted into the SalI and BamHI sites of pEGFP-C1 for the construction of plasmid pEGFP-STAT4. The DNA fragment of STAT4 with the deletion of amino acids 395-416 was amplified by RCR and introduced into the SalI and BamHI sites of pEGFP-C1 which was named pEGFP-STAT4-Del. Classic NLS DNA sequence of SV40 T antigen was inserted into the XhoI and HindIII sites of pEGFP-C1. This plasmid was named as pEGFP-NLS and used as a positive control. Plasmid pEGFP-NLS-STAT4-Del was constructed by inserting STAT4-Del into SalI and BamHI sites of pEGFP-NLS. These plasmids were transiently transfected into Caski cells, respectively. The results showed that, after these transfected cells were stimulated by IL-12, wild type STAT4 existed in the cytoplasm at 0 min, and was predominantly localized to the nucleus at 45 min, and distributed in both cytoplasm and nucleus at 60 min, suggesting that STAT4 translocates from cytoplasm into nucleus and finally re-entries into the cytoplasm during the stimulation of IL-12. However, deletion mutant of STAT4 was arrested in cytoplasm during the IL-12 stimulation. Leptomycin B, which specifically blocks protein export from nucleus into cytoplasm, was used to further demonstrate whether STAT4-Del is transferred into nucleus even with stimulation of IL-12. After the transfected cells were pre-treated by leptomycin B, the wild type STAT4 was mainly localized in nucleus after the IL-12 stimulation, suggesting that STAT4 was translocated from cytoplasm into nucleus by the stimulation of IL-12. On the other hand, the deletion mutant of STAT4 distributed in cytoplasm throughout, implying that the mutant STAT4 lacking of amino acids 395-416 cannot move into nucleus. Furthermore, the insertion of classic NLS into EGFP-STAT4-Del restored nuclear import of STAT4-Del. These results suggest the amino acids 395-416 is a dsNLS mediating IL-12-stimulated nuclear import of activated STAT4.

Detection of STAT2 in early stage of cervical premalignancy and in cervical cancer

OBJECTIVE@#To measure the expression pattern of STAT2 in cervical cancer initiation and progression in tissue sections from patients with cervicitis, dysplasia, and cervical cancer.@*METHODS@#Antibody against human STAT2 was confirmed by plasmids transient transfection and Western blot. Immunohistochemistry was used to detect STAT2 expression in the cervical biopsies by using the confirmed antibody against STAT2 as the primary antibody.@*RESULTS@#It was found that the overall rate of positive STAT2 expression in the cervicitis, dysplasia and cervical cancer groups were 38.5%, 69.4% and 76.9%, respectively. The STAT2 levels are significantly increased in premalignant dysplasia and cervical cancer, as compared to cervicitis (P< 0.05). Noticeably, STAT2 signals were mainly found in the cytoplasm, implying that STAT2 was not biologically active.@*CONCLUSIONS@#These findings reveal an association between cervical cancer progression and augmented STAT2 expression. In conclusion, STAT2 increase appears to be an early detectable cellular event in cervical cancer development.

Comparative study on the effect of restrictive fluid management strategy on the early pulmonary function of patients with severe burn / 中华烧伤杂志

<p><b>OBJECTIVE</b>To retrospectively analyze the effect of restrictive fluid management strategy (RFMS) on the early pulmonary function and the prognosis of patients with extremely severe and extensive burn.</p><p><b>METHODS</b>Thirteen patients with extremely severe burn hospitalized from June 2010 to November 2011, being treated with RFMS in the fluid reabsorption stage, were enrolled as treatment group. Twenty-six patients with extremely severe burn hospitalized from March 2008 to November 2011, being treated with normal fluid therapy in the fluid reabsorption stage, were enrolled as control group. The match proportion between treatment group and control group was 1:2. Fluid intake, fluid output, fluid balance (the difference between fluid intake and output), and plasma albumin level from post burn day (PBD) 3 to 10, pulmonary oxygenation index on PBD 3, 5, 7, 10, and 14, occurrence of lung and blood stream infections from PBD 7 to 14, and occurrence of acute respiratory distress syndrome (ARDS), occurrence of other organ complications, and mortality within 2 weeks post burn (PBW) were recorded and compared. Measurement data were processed with t test and randomized blocks analysis of variance, enumeration data were processed with Fisher's exact test.</p><p><b>RESULTS</b>Daily fluid intake of patients showed a tendency of decrease in both groups from PBD 3 to 10. Except for that of PBD 4, there was no statistically significant difference between two groups in fluid intake (with F values from 0.072 to 1.939, P values all above 0.05). Daily fluid output of patients showed a tendency of increase in both groups from PBD 3 to 10. It peaked on PBD 10 in control group and PBD 6 in treatment group. The mean daily fluid output was higher in treatment group than in control group from PBD 4 to 9, but without statistically significant difference (with F values from 0.001 to 3.026, P values all above 0.05). Fluid balance lowered in both groups, and it was the lowest on PBD 10 in control group and PBD 6 in treatment group. Fluid balance was lower in treatment group than in control group from PBD 3 to 7, and it showed statistically significant differences on PBD 4, 5, and 6 (with F values from 4.799 to 8.031, P values below 0.05). Plasma albumin level was higher in treatment group than in control group from PBD 3 to 10, with statistically significant differences observed on PBD 4, 9, and 10 (with F values from 5.691 to 10.551, P < 0.05 or P < 0.01). Pulmonary oxygenation index was higher in treatment group than in control group from PBD 3 to 14, with statistically significant differences observed on PBD 7 (respectively 372 ± 78 in treatment group and 291 ± 92 in control group, F = 5.184, P < 0.05) and 14 (respectively 354 ± 39 in treatment group and 283 ± 72 in control group, F = 8.683, P < 0.05). Lung infection and blood stream infection were respectively observed in 1 and 4 patient (s) in treatment group, and 9 and 11 patients in control group from PBD 7 to 14. Occurrence of ARDS, occurrence of other organ complications, and mortality were fewer in treatment group than in control group within PBW 2, though the differences were not statistically significant (P values all above 0.05).</p><p><b>CONCLUSIONS</b>RFMS is a useful strategy in improving early pulmonary oxygenation of patients with extremely severe and extensive burn by promoting the process of fluid reabsorption and rebalance. This strategy may be also beneficial for the prevention of organ complications as well as a better prognosis in severely burned patients.</p>

Protective effect of ginsenoside Re on myocardial cells of neonatal SD rat subjected to hypoxia injury / 中华烧伤杂志

<p><b>OBJECTIVE</b>To investigate the protective effect of ginsenoside Re on myocardial cells of neonatal SD rat with hypoxia injury, and to explore its mechanism.</p><p><b>METHODS</b>The primary passage of myocardial cells collected from neonatal SD rats were divided into A group (with ordinary treatment), B group [exposed to hypoxia (1% O2, 5% CO2, 94% N(2)) for 12 hours after being cultured for 48 hours], C group (pretreated with 80 g/L ginsenoside Re for 30 minutes after 48 hours of ordinary culture, then exposed to hypoxia for 12 hours), D group (received the same treatment as used in C group except for using 40 g/L ginsenoside Re), E group (received the same treatment as used in C group except for using 20 g/L ginsenoside Re) according to the random number table, with 6 samples in each group. Myocardial cell supernatants were collected for determination of content of lactate dehydrogenase (LDH) with enzyme linked immunosorbent assay. Fluorescence recovery after photobleaching technique was used to detect gap junction intercellular communication (GJIC). Result was observed by laser scanning confocal microscope. Data were processed with paired t test.</p><p><b>RESULTS</b>(1) Compared with that in B group [(403 ± 22) U/L], contents of LDH in E, D, and C groups were obviously decreased [(255 ± 16), (241 ± 13), (237 ± 24) U/L, with t value respectively 5.1, 5.2, 8.3, P values all below 0.05]. (2) The fluorescence recovery rate in A group was (74.8 ± 3.6)% 10 min after quenching, which was higher than that in B group [(13.2 ± 5.6)%, t = 15.2, P < 0.01]. The fluorescence recovery rate in C, D, and E groups was respectively (39.5 ± 2.9)%, (36.2 ± 3.1)%, and (34.3 ± 3.9)% 10 min after quenching, all higher than that in B group (with t value respectively -6.6, -41.9, 18.3, P values all below 0.05).</p><p><b>CONCLUSIONS</b>Ginsenoside Re pretreatment, particularly with a dose of 20 g/L, can protect myocardial cells from hypoxia injury, and the effect may be attributable to inhibition of release of LDH and improvement of the GJIC function.</p>

Qualitative analysis of the main chemical constituents of shuanghuanglian injection powder and their origin by HPLC-ESI/MS/MS spectrometry / 药学学报

This paper is to report the analysis of the main chemical constituents of Shuanghuanglian injection powder and determination of their origin. The sample solution was analyzed by a Zorbax C18 column with a gradient mobile phase comprised of methanol and 0.25% acetic acid solution. Both UV and electrospray ionization mass spectrometry detector were used simultaneously, -Q1-scan detection mode was evaluated for the identification of the LC peaks. To analyze the mass spectrum of every LC peaks, 43 molecular mass from the ion chromatogram of Shuanghuanglian injection powder were identified and among them, structure of 20 compounds were elucidated, and the data were sorted to the three component herbs, separately.

A clinical study of fungal infection in burn patients / 中华烧伤杂志

<p><b>OBJECTIVE</b>To address the features of the fungal infection after burn injury in clinic.</p><p><b>METHODS</b>Three thousand nine hundred and nine burn patients admitted to our institute from Jan. 2003 to Dec. 2006 were involved in this study. Two thousand two hundred and seventy-one samples were harvested for fungal detection by culture from 467 patients suspected to be infected by fungi based on their clinic manifestations. The collected samples included wound tissue, blood, urine, stool, sputum, catheters and others. The antibiotic sensitivity of the identified fungi were determined by routine method. When same kind of fungus was found from different samples taken from one patient, it was recorded as one positive sample. The samples were ranked in an ascending order as wound secretion, stool, urine, sputum and bronchial alveolar lavage fluid, arteriovenous catheter or urinary catheter, blood. Only the positive sample of the highest rank source was recorded as the positive strain of fungus from this particular patient.</p><p><b>RESULTS</b>It was found 61 fungal positive samples from the 2271 samples collected. Out of 467 patients, 38 strains of fungi were detected from 36 burn patients during the investigated period, the incidence was 0.92% (36/3909). The most three commonest types among the identified 38 strains of fungi were Candida tropicalis (42.1%), Candida albicans (31.6%) and Candida famata (T. Famata, 10.5%). The drug sensitivity tests demonstrated that most of the strains detected in this investigation, with the exception of candida glabrata, were sensitive to most of the routine antimycotics agents such as Amphotericin B, Fluconazole, and Itraconazole etc. Among the 36 fungus positive patients, in 18 patients the burn area exceeded 80% TBSA, 12 patients with 50%-79% TBSA, 4 patients with 30%-49% TBSA, and in 2 patients the burn area was smaller than 30% TBSA. It was found most of the fungal infections (77.78%) occurred 2 weeks after burn injury, and 8 of the 36 fungus-infected patients died (the mortality was 22.22%). Conclusions Further examinations are necessary to confirm the diagnosis in burn patients suspected to have fungal infection. Once fungal infections are confirmed, antimycotic therapy must be started immediately.</p>

Clinical practice and evaluation of relative fluid resuscitation formula at burn shock stage / 中华烧伤杂志

<p><b>OBJECTIVE</b>To evaluate the application of the Third Military Medical University (TMMU) formula for fluid resuscitation on the major burn patients during shock stage.</p><p><b>METHODS</b>Seventy-one thermal injury patients (burn area more than 30% TBSA, without especial illness, hospitalization within 8 hour after burn) admitted from 2005 to 2007 were divided into adult group (n = 46), child group (n = 25). Fluid resuscitation was initiated as per the TMMU formula.</p><p><b>RESULTS</b>All patients survived the first 48 hours post burn injury and none developed recognized complications associated with fluid resuscitation. The average infused fluid was 16% approximately 38% more than the calculated in both adult and child groups. The average urine output during the first 24 hours post burn injury was 1.1 approximately 1.2 mL x kg(-1) x h(-1) in the two groups, but reached 1.2 mL and 1.7 mL x kg(-1) x h(-1) during the second 24 hours in adult and child groups respectively.</p><p><b>CONCLUSION</b>TMMU formula for fluid resuscitation is a feasible option for major burn patients. Individual fluid resuscitation, guided by the physiological response, is also important and necessary.</p>

Association of HLA-A, B and DRB1 alleles with leukemia in Han population in Hunan Province / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the association of HLA-A, B, and DRB1 alleles with leukemia in the Han population in Hunan Province.</p><p><b>METHODS</b>HLA-A, B, and DRB1 alleles were genotyped in 105 patients with chronic myelocytic leukemia, 25 with acute lymphocytic leukaemia, and 48 with acute nonlymphocytic leukemia using polymerase chain reaction with sequence-specific primer (PCR-SSP). The hemopoietic stem cells from 3,664 unrelated normal individuals of Han nationality in Hunan were used as the control group.</p><p><b>RESULTS</b>The phenotypic frequencies of HLA-B58, DR12, and DR14 were significantly higher in patients with chronic myelocytic leukemia than in the control group, with relative risk of 6.1287, 1.6519, and 1.6479, respectively. In patients with acute lymphocytic leukaemia, the phenotypic frequency of HLA-B58 was significantly higher than that in the control group, with the relative risk of 7.4055. In patients with acute nonlymphocytic leukemia, the frequencies of HLA-B58 and DR8 phenotypes were significantly higher but HLA-A24 frequency was significantly lower than those of the control group, with the relative risk of 13.9789, 2.2839, and 0.4012, respectively.</p><p><b>CONCLUSION</b>HLA-B58, DR12, DR14 alleles appear to contribute to the genetic susceptibility of patients with chronic myelocytic leukemia. HLA-B58 allele can be associated with the genetic susceptibility for patients with acute lymphocytic leukaemia. In patients with acute nonlymphocytic leukemia, HLA-B58 and DR8 are probably the susceptible alleles whereas HLA-A24 allele may play a protective role.</p>

Correlation between MHC class I-related chain A gene *008 allele and human cytomegalovirus infection / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the correlation between MHC class I-related chain A (MICA) gene *008 allele and human cytomegalovirus (HCMV) infection.</p><p><b>METHODS</b>MICA*008 allele was detected in 86 patients with chronic granulocytic leukemia and 81 unrelated normal individuals by way of sequence-specific primers (PCR-SSP). Anti-HCMV IgM was also detected in the sera of these subjects with enzyme linked immunosorbent assay (ELISA).</p><p><b>RESULTS</b>MICA*008 allele frequency was lower in patients with chronic granulocytic leukemia than in the control group (22.2% vs 34.3%, Chi(2)=4.98, P<0.05). The infection rate of HCMV was significantly higher in those individuals with genotype of MICA*008 (-) than in those with MICA*008 (+), and moderate correlation was suggested between MICA*008 and HCMV infection (C=0.5829, 0.6142).</p><p><b>CONCLUSION</b>Individuals with MICA*008 positivity is not liable to HCMV infection, but those with MICA*008 (-) can be vulnerable to HCMV infection, suggesting an inverse correlation between MICA*008 allele with HCMV.</p>

MICA*008/A5.1 allele and HCMV infection in kidney transplanted donees of Hunan Han nationality / 中南大学学报(医学版)

OBJECTIVE@#To investigate the relationship between MICA*008/A5.1 allele and human cytomegalovirus (HCMV) infection in kidney transplanted donees of Hunan Han nationality.@*METHODS@#The MICA*008/A5.1 allele based on 91 kidney transplanted donees and 81 unrelated normal individuals of Han nationality in Hunan Province were analyzed by PCR/SSP assay. At the same time, anti-HCMV antibody IgM was detected in the serum by ELISA method.@*RESULTS@#The positive rate of MICA*008/A5.1 allele was significantly higher in the control group (56.79%) than that in the kidney transplanted donee group (34.07%) (P <0.05). The infection rate of HCMV in those individuals whose genotype was MICA*008/A5.1 (-) was significantly higher than that in the MICA*008/A5.1(+).@*CONCLUSION@#The individual whose genotype is MICA*008/A5.1 (+) is not liable to HCMV infection, but the individual whose genotype is MICA*008/A5.1 (-) is liable to HCMV infection.

Multi-center clinical study of acticoat (nanocrystalline silver dressing) for the management of residual burn wounds / 中华烧伤杂志

<p><b>OBJECTIVE</b>To investigate the clinical efficacy and safety of Acticoat (nanocrystalline silver dressing) for the treatment of residual burn wounds.</p><p><b>METHODS</b>Ninety-eight patients with 166 residual burn wounds were enrolled in the multi-center randomized clinical trials. In addition to the routine treatment, Acticoat was applied onto the wounds of the trial group once a day if there was much exudation from the wound, or the dressing change was made every other two days when the wounds were clean. Silver sulfadiazine (SD-Ag) was used in the control group of patients. The healing time was observed up to 20 days. The healing rate on the 15th day after treatment was taken as the percentage of healing.</p><p><b>RESULTS</b>The average healing time was (12 +/- 5) days after the application of Acticoat, which was significantly shorter than that in control wounds with SD-Ag (16 +/- 6) days, (P = 0.005 < 0.01). The total effective rate of the wounds for trial was 97.05%, which was higher than that in control (94.17%) group, but there was no statistically significant difference. The bacterial clearing rate of the Acticoat group on the 6th and 12th post treatment day was 21.7% and 43.5% respectively, which was significantly higher than that in control group. No side-effect was observed in the two groups during the study.</p><p><b>CONCLUSION</b>Acticoat with nanocrystalline silver can promote the healing of residual burn wounds effectively.</p>

High-performance liquid chromatography-electrospray ionization/tandem mass spectrometry of isofraxidin in Sarcandra glabra / 南方医科大学学报

<p><b>OBJECTIVE</b>To establish a method for determining isofraxidin concentrations in Sarcandra glabra and Qingrexiaoyanning capsules with high-performance liquid chromatographic-mass spectrometry.</p><p><b>METHODS</b>Isofraxidin was extracted from Sarcandra glabra and Qingrexiaoyanning capsules with acetic ether and chloroform, respectively, and separated by isocratic reversed-phase chromatography. The mass spectrometric system was operated in multiple reaction monitoring mode. A pair of ions: precursor ion m/z 223 with product ion m/z 162 were chosen for the quantification of the analyte.</p><p><b>RESULTS</b>The retention time of isofraxidin was 6.60 min, and the calibration curve was linear over a concentration range from 484 to 9 680 ng/ml. The average recovery was 96.7% and RSD 4.49%, with detection limit of 1 ng/ml.</p><p><b>CONCLUSION</b>The method is rapid, selective and sensitive for determining isofraxidin in Sarcandra glabra and Qingrexiaoyanning capsules.</p>

Polymorphism of short tandem repeat of exon 5 of MHC class-I chain related gene A and association with nasopharyngeal carcinoma in a southern Chinese population / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To explore the association between the short tandem repeat polymorphism of exon 5 of MICA gene (MICA-STR) and nasopharyngeal carcinoma (NPC) in a southern Chinese population.</p><p><b>METHODS</b>One hundred and twenty-seven consecutive NPC patients and 112 randomly selected normal controls residing in southern China mainland were analyzed for MICA-STR allelic variation and MICA gene deletion by fluorescent polymerase chain reaction-gene scanning and polymerase chain reaction-sequence specific priming.</p><p><b>RESULTS</b>MICA*A9 was observed at significantly higher frequency in the NPC patient group than in the control group (relative risk = 2.524, P = 0.001,Pc = 0.006); whereas MICA*A5.1 was present at significantly lower frequency in the NPC patient group than in the control group (RR = 0.418, P = 0.0004, Pc = 0.0026). Further analysis revealed that MICA*A9 was over-represented in male NPC patients, compared with male controls (RR = 3.23, P = 0.00095, Pc = 0.006); whereas MICA*A5.1 was present at significantly lower frequency in male NPC patients, compared with male controls (RR = 0.372, P = 0.0007, Pc = 0.004). None of the MICA-STR variants showed statistically significant frequency difference between female NPC patients and female controls (Pc > 0.05).</p><p><b>CONCLUSION</b>MICA-STR polymorphism is associated with NPC, and MICA*A9 is a genetic susceptibility marker of male individuals for NPC in a southern Chinese population.</p>

Biological characteristics of human keratinocytes cultured in serum-free medium without feeder layer / 中华烧伤杂志

<p><b>OBJECTIVE</b>To establish an optimal method for serum-free and feeder layer-free culture of human keratinocytes and to investigate their biological characteristics.</p><p><b>METHODS</b>The keratinocytes were harvested from human foreskin of 5 children (aged 5-10 yr) and 5 adults (aged 20-30 yr). The samples were isolated by two-step digestion and the quantities of primary harvested HKCs were determined. The HKCs were then cultured in KCS serum-free culture medium. The morphology of HKCs were observed under light microscope. The HKCs and their growing speed were observed and identified under fluorescent microscope. The growth curve of HKCs was detected with MTT method, and the cell cycle was determined with flow cytometry.</p><p><b>RESULTS</b>The number of harvested HKCs from children [(1.780 +/- 0.010) x 10(6)/cm(2)] was obviously higher than that from adults [(1.490 +/- 0.120) x 10(6)/cm(2)], (P < 0.01). Freshly isolated primary HKCs were round and transparent, and 94% of them were trypan blue resistant. The adherent speed and rate and lucent degree of multiply passaged HKCs increased followed by each passage. Under the fluorescent microscope, the cells exhibited strong Kelly fluorescence in the cytoplasm and with no staining in the nucleolus, thus the cells were identified as HKCs. The HKCs from children for skin could be passaged for more times [(11.0 +/- 1.2) times] than that from adults [(9.2 +/- 0.8) times], (P < 0.05). There was no clear sign of incubation period in the growth curve of HKCs, and both cellular proliferating speed and rate of proliferation were high. The percentage of cells in G1, G2 and S phase and the proliferation index was 36.15%, 25.17%, 38.68% and 63.85%, respectively.</p><p><b>CONCLUSION</b>Serum-free and feeder layer-free culture seems to be an ideal method for the cultivation of HKCs.</p>

Establishment of a canine model of severe smoke inhalation injury on unilateral lung / 中华烧伤杂志

<p><b>OBJECTIVE</b>To establish a canine model of severe smoke inhalation injury on unilateral lung, in order to observe the pathomorphological changes in the injured lung within 24 postburn hours (PBHs).</p><p><b>METHODS</b>Twenty five mongrel dogs were employed in the study and randomized into 3 groups. The left lung was injured by inhaling smoke produced by burning sawdust with sparing the right lung with a breathing tube in 10 dogs in group A. A conventional model of smoke inhalation injury to bilateral lungs was reproduced in 8 dogs in group B, and dogs in group C not subjected to smoke inhalation served as controls. Hemodynamic changes, blood gas analysis and the pathophysiologic changes in the lungs were observed within 24 PBHs.</p><p><b>RESULTS</b>All of the dogs in groups A and C survived. Hemodynamic indices in the dogs in groups A and C remained stable without showing signs of systemic hypoxia. The arterial oxygen partial pressure in dogs of group A was 65 +/- 5 mm Hg, and the oxygen saturation in the mixed blood was 0.64 +/- 0.04 at 24 PBHs, and they were much lower than those in group C but higher than those in group B. The pathological changes in the injured side of the lungs in group A were similar to those in group B with high consistency, and the changes, though milder, could also be identified in the contralateral uninjured lung. Five dogs died in the group B within 24 hours after smoke inhalation and the survivors showed signs of multiple organ failure.</p><p><b>CONCLUSION</b>The canine model of acute severe unilateral pulmonary smoke inhalation injury was reproduced reliably, and could be an ideal model for the study on smoke inhalation injury.</p>

Studies on design, synthesis and biodegradation of carrier for colon-site specific, drug delivery system / 药学学报

<p><b>AIM</b>To design and synthesize a novel vector for colon-site specific drug delivery system and investigate the relationship between the biodegradation properties and composition of materials in the simulated colon fluid.</p><p><b>METHODS</b>The azocopolymer P (HEMA-MMA-MAA) was synthesized using 2-hydroxyethylmethacrylate (HEMA), methyl methacrylate (MMA) and methacrylic acid (MAA) as comonmer, azobisisobutyronitrilel (AIBN) as initiator, cross-linked with divinylazobezene (DVAB). The chemical structure of the synthesized series of azocopolymer is examined by UV, FTIR spectroscopy and nuclear magnetic resonance data. Their swelling behavior is evaluated by the swelling equilibrium parameter Q, the biodegradation tests of the materials were carried out at physiologically relevant buffer designed to mimic the colon environment. The biodegradation properties were assessed using the differential scanning calorimeters (DSC) and gel permeation chromatography (GPC) and the morphology on the surface of materials before and after degradation was observed by scanning electron microscopy (SEM).</p><p><b>RESULTS</b>The swelling equilibrium parameter Q increased with increasing the contents of HEMA and MAA in the materials. The degradation behavior was relevant to the ratio of three components in the copolymers.</p><p><b>CONCLUSION</b>This materials may become a good carrier for the colon-site specific drug delivery system if the contents of commoners HEMA, MMA and MAA are adjusted reasonably.</p>

Changes in the polymorphonuclear leukocyte infiltration, hemorrhage and plasma exudation in the pulmonary tissue in rats with smoke inhalation injury / 中华烧伤杂志

<p><b>OBJECTIVE</b>To investigate the different velocity of polymorphonuclear neutrophil (PMN) infiltration, erythrocyte diapedesis and plasma exudation into the pulmonary tissue of the rats inflicted with smoke inhalation injury, so as to explore the different mechanisms of their existence in rat pulmonary tissue after inhalation injury.</p><p><b>METHODS</b>The rat smoke inhalation injury model was employed in the study. Wistar rats were inflicted with smoke inhalation injury, and then sacrificed at 1, 3, 6, 12 and 24 post injury hours (PIH). 131I-BSA, 99mTc-PMN or 99mTc-erythrocytes (RBC) were injected into rat pulmonary tissue 1 hour before sacrifice. Isotonic saline was infused into blood vessel to wash out circulation blood. Then the pulmonary tissue samples were harvested for gamma-value counting and then weighed. The infiltration of 131I-BSA, 99mTc-PMN or 99mTc-RBC in pulmonary tissue per gram and per minute was calculated, and MPO content was measured by phosphate T-tolidine method.</p><p><b>RESULTS</b>The amount of RBC diapedesis in rat lung tissue peaked at 1 PIH, decreased thereafter and approached to normal level at 24 PIH. The amount of PMN infiltration increased at 3 PIH, slightly decreased at 6 PIH but still higher than that in normal tissue, and increased again at 24 PIH. The pulmonary tissue content of MPO gradually increased from 1 PIH to 24 PIH. The pulmonary tissue content of 131I-BSA began to increase at 1 PIH and peaked at 6 PIH, and remained higher than that in normal tissue till 24 PIH.</p><p><b>CONCLUSION</b>Even though there was remarkable postburn increase in the erythrocyte diapedesis, neutrophil infiltration and albumin exudation with different peak time points (1, 3 and 6 PIH, respectively), Inflammation seemed not to be the premise of erythrocyte diapedesis, while the secondary inflammatory reaction might be the main cause of pulmonary edema.</p>

Pathomorphologic changes in rat lungs after being perfused with the bronchoalveolar lavage fluid from the dogs inflicted by smoke inhalation injury at early post-injury stage / 中华烧伤杂志

<p><b>OBJECTIVE</b>To investigate the biological activities of the bronchoalveolar lavage fluid (BALF) harvested from dogs with smoke inhalation injury at early post injury stage.</p><p><b>METHODS</b>BALF was harvested from normal dogs and those inflicted by smoke inhalation injury for the employment in the study. Ninety four Wistar rats were employed in the study and were randomly divided into A (n=28), B (n=29) and C (n=37) groups. The rats were perfused intra-tracheally with normal saline, BALF from normal dog and BALF from injured dog, respectively in A, B and C groups. Every 7 rats in each group were used before the perfusion as normal control. The rats in control group and those in A, B and C groups at 4, 12 and 24 hours after BALF perfusion were sacrificed. The survival rate and the pathomorphological gross and microscopic changes in pulmonary tissue in all groups were observed. In addition, the contents of prostaglandin F1alpha/thromboxane B2 (PGF1alpha/TXB2), tumor necrosis factor (TNF-alpha) and myeloperoxidase (MPO) in pulmonary tissue homogenization and the pulmonary capillary permeability were determined simultaneously.</p><p><b>RESULTS</b>All the rats in A and B groups survived, but nine in C group died before sacrificing. The BALF from dogs inflicted by smoke inhalation injury could cause rat lung exhibiting pathomorphological changes similar to those in rats inhaling smoke directly. The rat pulmonary tissue contents of PGF1alpha/TXB2 in A and B groups after the perfusion tended to increase, while that in C group decreased gradually (P <0.01). The rat pulmonary tissue contents of TNF-alpha and MPO content in A and B groups after the perfusion revealed no obvious change (P >0.05), while those in C group increased evidently at 4 hour after the perfusion and decreased thereafter (P <0.05-0.01). The pulmonary capillary permeability in C group was higher than that in A and B groups at 4 hour after the perfusion (P < 0.01).</p><p><b>CONCLUSION</b>The BALF from canine lungs during the early stage of inhalation injury exhibited biological activities. The primary and secondary pulmonary injury could be prevented or ameliorated by massive pulmonary lavage during early post injury stage after smoke inhalation injury.</p>