Some thoughts on the research of mesenchymal stem cell exosomes and wound microenvironment / 中华烧伤杂志

Since researchers have found that the conditioned medium and exosomes of mesenchymal stem cells (MSCs) had the biological effects equivalent to those of MSCs, MSC exosomes (MSC-Exos), the representative product of MSCs' paracrine effect, have become the research focus of the "cell-free" therapy of MSCs. However, most researchers currently use conventional culture condition to culture MSCs and then isolate exosomes for the treatment of wound or other diseases. Theoretically, the paracrine effect of MSCs is directly associated with the pathological condition of the wound (disease) microenvironment or in vitro culture condition, and their paracrine components and biological effects may be altered with the changes of the wound (disease) microenvironment or in vitro culture condition. Thus, the feasibility of using traditional culture condition to culture MSCs for exosome extraction for the treatment of different diseases without considering the actual situation of the disease to be treated needs further discussion. Therefore, the author suggests that the research of MSC-Exos should consider the microenvironment of the wound (disease) to be treated. as much as possible, otherwise the extracted MSC-Exos may not be "accurate" or may not really achieve the treatment effect of MSCs. In this article, we summarized some thoughts of the author and problems related to the researches about MSC-Exos and wound microenvironment, and hoped to discuss with researchers.

Clinical application effects of two longitudes three transverses method in perforator location of thoracodorsal artery perforator flap and deep wound repair / 中华烧伤杂志

Objective: To explore the clinical application value of two longitudes three transverses method in the location of the perforator of thoracodorsal artery perforator and deep wound repair. Methods: The retrospectively observational study was conducted. From December 2018 to June 2020, 17 patients with deep wounds who were admitted to the Affiliated Hospital of Zunyi Medical University met the inclusion criteria and were included in this study, including 7 males and 10 females, aged 12 to 72 years. The wound areas of patients after debridement were 7 cm×3 cm to 11 cm×7 cm. Two longitudinal lines were located through the midpoint of the armpit, the posterior superior iliac spine, and the protruding point of the sacroiliac joint, and three transverse lines were located 5, 10, and 15 cm below the midpoint of the armpit between the two longitudinal lines, i.e. two longitudes three transverses method, resulting in two trapezoidal areas. And then the thoracodorsal artery perforators in two trapezoidal areas were explored by the portable Doppler blood flow detector. On this account, a single or lobulated free thoracodorsal artery perforator flap or flap that carrying partial latissimus dorsi muscle, with an area of 7 cm×4 cm to 12 cm×8 cm was designed and harvested to repair the wound. The donor sites were all closed by suturing directly. The number and location of thoracodorsal artery perforators, and the distance from the position where the first perforator (the perforator closest to the axillary apex) exits the muscle to the lateral border of the latissimus dorsi in preoperative localization and intraoperative exploration, the diameter of thoracodorsal artery perforator measured during operation, and the flap types were recorded. The survivals of flaps and appearances of donor sites were followed up. Results: The number and location of thoracodorsal artery perforators located before operation in each patient were consistent with the results of intraoperative exploration. A total of 42 perforators were found in two trapezoidal areas, with 2 or 3 perforators each patient. The perforators were all located in two trapezoid areas, and a stable perforator (the first perforator) was located and detected in the first trapezoidal area. There were averagely 1.47 perforators in the second trapezoidal area. The position where the first perforator exits the muscle was 2.1-3.1 cm away from the lateral border of the latissimus dorsi. The diameters of thoracodorsal artery perforators were 0.4-0.6 mm. In this group, 12 cases were repaired with single thoracodorsal artery perforator flap, 3 cases with lobulated thoracodorsal artery perforator flap, and 2 cases with thoracodorsal artery perforator flap carrying partial latissimus dorsi muscle. The patients were followed up for 6 to 16 months. All the 17 flaps survived with good elasticity, blood circulation, and soft texture. Only linear scar was left in the donor area. Conclusions: The two longitudes three transverses method is helpful to locate the perforator of thoracodorsal artery perforator flap. The method is simple and reliable. The thoracodorsal artery perforator flap designed and harvested based on this method has good clinical effects in repairing deep wound, with minimal donor site damage.

Effect of Modified Guipitang Combined with Xingnao Kaiqiao Acupuncture in Treatment of Heart and Spleen Insufficiency Syndrome of Non-dementia Vascular Cognitive Impairment / 中国实验方剂学杂志

Objective: To explore the curative effect and partial mechanism of modified Guipitang combined with Xingnao Kaiqiao acupuncture in the treatment of non-dementia vascular cognitive impairment (VCIND). Method: Totally 122 patients with VCIND admitted to the First Teaching Hospital of Tianjin University of Traditional Chinese Medicine (TCM) from May 2017 to October 2018 were randomly divided into modified decoction group (39 cases), acupuncture group (42 cases) and combination group (41 cases). All of the three groups were orally given routine anticoagulants and lipid-lowering drugs. The decoction group was orally given modified Guipitang 150 mL/times, 2 times/day, in addition to the routine treatment, the acupuncture group was treated with Xingnao Kaiqiao acupuncture method in addition to the routine treatment, involving Shuigou, Neiguan (bilateral), Sanyinjiao (bilateral), Sishencong, Xuanzhong (bilateral) and Taixi (bilateral) acupoints, 2 times/day, for six days a week, the combined group was treated with Xingnao Kaiqiao acupuncture in addition to modified Guipitang. All of the three groups were treated for 8 weeks. The Montreal cognitive assessment scale (MoCA scale, Beijing version) and activity of daily life (ADL) scale, TCM symptoms and clinical efficacy were scored before treatment, 4 weeks after treatment and 8 weeks after treatment in three groups. Serum levels of calcitonin gene-related peptide (CGRP) and cone-like protein-1 (VILIP-1) were measured by enzyme-linked immunosorbent assay at different time points. Result: Compared with decoction group and acupuncture group, MoCA score, ADL score and TCM syndrome score of the combined group were decreased, the total effective rate was increased significantly after 4 and 8 weeks of treatment, the serum CGRP content was increased, and the VILIP-1 content was decreased. Conclusion: Modified Guipitang combined with Xingnao Kaiqiao acupuncture has a definite curative effect on VCIND with heart and spleen insufficiency syndrome. Its mechanism may be related to the expansion of blood vessels, the alleviation of blood supply of brain and the reduction of neuron injury.

Effects of Polygonum cuspidatum on AMPK-FOXO3α Signaling Pathway in Rat Model of Uric Acid-Induced Renal Damage / 中国结合医学杂志

BACKGROUND@#To observe the effects of Chinese medicine (CM) Polygonum cuspidatum (PC) on adenosine 5'-monophosphate-activated protein kinase (AMPK), forkhead box O3α (FOXO3α), Toll-like receptor-4 (TLR4), NACHT, LRR and PYD domains-containing protein 3 (NLRP3), and monocyte chemoattractant protein-1 (MCP-1) expression in a rat model of uric acid-induced renal damage and to determine the molecular mechanism.@*METHODS@#A rat model of uric acid-induced renal damage was established, and rats were randomly divided into a model group, a positive drug group, and high-, medium-, and low-dose PC groups (n=12 per group). A normal group (n=6) was used as the control. Rats in the normal and model groups were administered distilled water (10 mL•kg) by intragastric infusion. Rats in the positive drug group and the high-, medium-, and low-dose PC groups were administered allopurinol (23.33 mg•kg), and 7.46, 3.73, or 1.87 g•kg•d PC by intragastric infusion, respectively for 6 to 8 weeks. After the intervention, reverse transcription polymerase chain reaction, Western blot, enzyme linked immunosorbent assay, and immunohistochemistry were used to detect AMPK, FOXO3α, TLR4, NLRP3, and MCP-1 mRNA and protein levels in renal tissue or serum.@*RESULTS@#Compared with the normal group, the mRNA transcription levels of AMPK and FOXO3α in the model group were significantly down-regulated, and protein levels of AMPKα1, pAMPKα1 and FOXO3α were significantly down-regulated at the 6th and 8th weeks (P<0.01 or P<0.05). The mRNA transcription and protein levels of TLR4, NLRP3 and MCP-1 were significantly up-regulated (P<0.01 or P<0.05). Compared with the model group, at the 6th week, the mRNA transcription levels of AMPK in the high- and medium-dose groups, and protein expression levels of AMPKα1, pAMPKα1 and FOXO3α in the high-dose PC group, AMPKα1 and pAMPKα1 in the mediumdose PC group, and pAMPKα1 in the low-dose PC group were significantly up-regulated (P<0.01 or P<0.05); the mRNA transcription and protein levels of TLR4 and NLRP3 in the 3 CM groups, and protein expression levels of MCP-1 in the medium- and low-dose PC groups were down-regulated (P<0.01 or P<0.05). At the 8th week, the mRNA transcription levels of AMPK in the high-dose PC group and FOXO3α in the medium-dose PC group, and protein levels of AMPKα1, pAMPKα1 and FOXO3α in the 3 CM groups were significantly up-regulated (P<0.01 or P<0.05); the mRNA transcription levels of TLR4 in the medium- and low-dose PC groups, NLRP3 in the high- and low-dose PC groups and MCP-1 in the medium- and low-dose PC groups, and protein expression levels of TLR4, NLRP3 and MCP-1 in the 3 CM groups were down-regulated (P<0.01 or P<0.05).@*CONCLUSION@#PC up-regulated the expression of AMPK and its downstream molecule FOXO3α and inhibited the biological activity of TLR4, NLRP3, and MCP-1, key signal molecules in the immunoinflammatory network pathway, which may be the molecular mechanism of PC to improve hyperuricemia-mediated immunoinflflammatory metabolic renal damage.

Efficacy of autologous peripheral blood mononuclear cells in treatment of high radial nerve injury / 中国组织工程研究

OBJECTIVE: To investigate the efficacy of autologous peripheral blood mononuclear cells in the treatment of high radial nerve injury. METHODS: From April 2011 to September 2015, 12 cases of radial nerve injury in the middle arm were treated. Preoperatively peripheral blood mononuclear cells were mobilized, and then 15 mL of mononuclear cell suspension was prepared on the operation day. Radial nerves scheduled for anastomosis were surgically explored and subjected to end-end anastomosis using outer membrane suturing under microscope. The anastomotic site of the nerve was enveloped with gelatin sponge soaked with 5 mL of autologous peripheral blood mononuclear cell suspension. The remaining 10mL of cell suspension was used for a multi-point injection into the local muscles, 0.5 mL at each point. Thereafter, the deep fascia and the incision were sutured in sequence. Postoperative antibiotic treatment was used to prevent infection for 48 hours, and upper limb immobilization lasted for 4 weeks. Performance of rehabilitation exercise was guided. During the follow-up, wrist dorsal extension and muscle strength of extensor carpi ulnaris and extensor digitorum communis were detected to evaluate the therapeutic efficacy. RESULTS AND CONCLUSION: All the patients were followed up for 15 to 36 months, with an average of 17 months. Efficacy was excellent in 9 cases, good in 2 cases, fair in 1 case and poor in 0 case. The excellent and good rate was 92%. The wrist dorsal extension could achieve the functional needs, and the thumb dorsal extension and finger extension basically met the functional requirements. It is suggested that autologous peripheral blood mononuclear cell transplantation can achieve good outcomes in the treatment of high radial nerve injury.

Establishment of stem cell curriculum and thinking of teaching practice in graduate education / 医学研究生学报

In order to provide reference for development of the new curriculum for graduate students "The principle,technical and clinical practice of stem cell" and improve the quality of teaching and learning,we summarized some experiences and lessons for this course.This paper introduced some measures and achievements on the teaching reform from several aspects including curriculum design,selection of teaching content,teaching idea and mode,training the teaching groups and so on.Moreover,it especially emphasized that teaching contents should not only pay attention to the basic and clinical application,but also combine the present laws and keep abreast of scientific and technological developments.In the process of teaching,we need to value the quality of teachers and improve teaching standards,meanwhile we should pay attention to the culture of thought and the capacity of innovation,stimulating students' interests in studies,improving autonomous study and practices ability through multiple means and channels.This course teaching got satisfactory results and the acquired practice experiences were worth of being popularized in the further graduate students teaching.

The research of fibroblasts from human hypertrophic scar showing a mesenchymal stem cell phenotype and multilineage differentiation potentialities / 中华整形外科杂志

<p><b>OBJECTIVE</b>To explore the CD phenotypic, protein expression and pluripotent differentiation of human hypertrophic scar fibroblasts cultured in vitro, so as to study the mechanisms of scar formation.</p><p><b>METHODS</b>Fibroblasts were isolated and cultured from human hypertrophic scar of 3 cases. The cells morphology was observed by inverted microscope, and the growing state of the third passage was detected by the cell counting meter of Vi-CELL. The cell surface markers CD105, CD14, CD73, CD34, CD44, CD45 and CD90 were identified by flow cytometry. The expression of CK19, Oct-4, Nanog and vimentin was detected by immunocytochemistry, and the expression of alpha-smooth muscle actin(alpha-SMA) was tested by immunofluorescence. The differentiated potential of fibroblasts of the third passage into adipogenic, osteogenic and chondrogenic lineages was assayed.</p><p><b>RESULTS</b>The primary passage fibroblasts showed the shape of spindle shaped or irregular polygon with a radiated or circinate of growing arrangement. The growth curve showed the cells growth was slow on the first and second day, and quick during the third to fifth day, which reached platform stage on the sixth or seventh day. The fibroblasts highly expressed mesenchymal stem cell surface markers-CD73, CD105, CD44, CD90, but not expressed hematopoietic stem cell surface markers-CD14, CD34, CD45 by flow cytometry. And positive expression of vimentin, Oct-4 and negative expression of CK19 were detected by Immunocytochemistry. Positive expression of alpha-SMA was also detected by immunofluorescence. Multidirectional differentiation induction indicated that the third passage cells could differentiate into adipogenic, osteogenic and chondrogenic lineages.</p><p><b>CONCLUSIONS</b>Human hypertrophic scar-derived fibroblasts show the biologic characteristics of mesenchymal stem cells, which may play an important role in wound healing and hypertrophic scar formation.</p>

An experiment study and clinical observation of the influence of free-skin-grafted scrotal avulsion injuries on spermatogenesis / 中华外科杂志

<p><b>OBJECTIVE</b>To explore the influence of Free-skin-grafted penoscrotal avulsion injuries on spermatogenesis.</p><p><b>METHODS</b>Forty-two male New Zealand albino rabbits during child-bearing period were divided into the experimental group (n = 24) and the control group (n = 18) using random digits table, and 24 female rabbits with reproductive history were used for mating experiment. The experimental group animal's scrotum skin were excised, and the split skin from abdominal region was used to repair the skin defect of scrotum. The control group did not any processing. Six rabbits were randomly chosen respectively in control group and on the 3rd and 8th weekend after the model was successfully established in experimental group. The testicular surface temperature was measured in the eighteen rabbits using the method of burying thermometer, then the testicular biopsy were performed for hematoxylin-eosin (HE) staining. On the 8(th) weekend after the model was successfully established in experimental group, matched-pair feed was performed in the other 12 rabbits respectively in experimental group and in control group. Observation of corresponding mother rabbit fertility. Three patients of penoscrotal avulsion injuries were treated using split skin grafts, and the information of sex life and the quality of sperm were obtained by follow up.</p><p><b>RESULTS</b>The testicular surface temperature was similar on the 3rd and 8th weekend after the model was successfully established in experimental group [(36.15 ± 0.24)°C, (36.77 ± 0.42)°C] with that of the control group. Testis tissue (HE) staining showed the tier of spermatogenic cells was rule arrangement and lot of mature sperms were found in the convoluted seminiferous tubules in control group. The tier of spermatogenic cells was diminished and disposed derangement, the spermatozoa were not seen on the 3(th) weekend of the experiment group. The tier of spermatogenic cells was increased and some spermatozoa were seen on the 8th weekend of the experiment group. Male and female matched-pair feed showed the experimental group conception rate 8/12, and 4.1 ± 3.2 rabbit babies were born averagely, while that of was 12/12 and 6.0 ± 1.3 in control group (P > 0.05). The skin grafts there were some contracture in early stage (1 - 2 months) when the skin grafts applied to repair the avulsing scrotum in three patients. But the skin grafts became loose with downward sagging and there were the good cosmetic result in one year, and without any contracture. The sperm quality was normal after the skin grafts applied to repair the avulsing scrotum in the late stage.</p><p><b>CONCLUSIONS</b>The skin grafting is little arrest the testicle spermatogenesis in the three methods (skin flap reconstruction scrotum, testicle buried, split skin grafting) that have usually been used to repair scrotum skin lose. For a young male, the best treatment for penoscrotal avulsion injuries is free skin grafting, while skin flaps are not recommended for reconstructing the scrotum.</p>

Effects of different ways in repairing scrotum of pigs with full-thickness burn on apoptosis and bcl-2 protein expression of spermatogenic cells / 中华烧伤杂志

<p><b>OBJECTIVE</b>To observe the effects of different ways in repairing scrotum of pigs with full-thickness burn on spermatogenesis of testis.</p><p><b>METHODS</b>Twenty male Guizhou miniature pigs were divided into normal control (NC), natural-healing (NH), flap-repairing (FR), and skin-grafting (SG) groups according to the random number table, with 5 pigs in each group. Pigs in NC group were not subjected to any injury. Scrotum of pigs in the latter three groups were inflicted with full-thickness burn. Wounds in NH group healed naturally. Wounds in FR group were repaired with inguinal region flap, and those in SG group with full-thickness skin from lower abdomen. Appearance of scrotum in the latter three groups was observed right after injury, and three months post injury or surgery (PIM or PSM). Specimens of testes of pigs in the latter three groups were obtained in PIM or PSM 3 to detect apoptosis of spermatogenic cells with TUNEL, and bcl-2 protein expression with immunohistochemistry. The same indexes were observed and determined in pigs of NC group. Data were processed with one-way analysis of variance and LSD test.</p><p><b>RESULTS</b>(1) Scrotum of pigs in NC group had skin folds with contraction function. Scrotum of pigs became hard with a leathery appearance right after burn in the other three groups. In PIM or PSM 3, wounds of pigs in NH group healed with scar, and the testes were squeezed into inguinal region. Scrotal skin of pigs in FR group was thick with testes in the scrotum, and that of pigs in SG group was thin with testes in the scrotum. (2) Spermatogenic cells in each level in NC group were arranged regularly, with few apoptotic spermatocytes and spermatoblasts. In NH, FR, and SG groups, seminiferous epithelium was thinner with most of the spermatogenic cells showing apoptosis, and they were mainly spermatogonia and spermatocytes. Apoptotic index of spermatogenic cells in NH, FR, SG, and NC groups was respectively (46.3 ± 3.3)%, (40.9 ± 3.5)%, (20.6 ± 2.3)%, (7.5 ± 1.9)%, and the difference among them was statistically significant (F = 405.65, P < 0.01). There were significant statistical differences among the former three groups (with P values below 0.01). (3) bcl-2 protein expression in NH, FR, SG, and NC groups was respectively (52 ± 5)%, (53 ± 4)%, (64 ± 5)%, (75 ± 5)%, and the difference among them was statistically significant (F = 56.63, P < 0.01). There was no significant statistical difference in bcl-2 expression between NH group and FR group (P = 0.66), and it was lower in both groups as compared with SG group (with P values below 0.01).</p><p><b>CONCLUSIONS</b>Either scar healing, flap transplantation, or SG in repairing scrotum with full-thickness burn in pigs inhibits spermatogenesis, but repair with SG produces less deleterious effect on the testis.</p>

Comparison of the short-term outcomes of surgical treatment for non-small cell lung cancer via video assisted thoracoscopic surgery and open thoracotomy / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To compare the short-term outcomes of surgical treatment for non-small cell lung cancer (NSCLC) by video-assisted thoracoscopic surgery (VATS) and open thoracotomy (OT).</p><p><b>METHODS</b>Data of 737 consecutive NSCLC patients who underwent surgical treatment for non-small cell lung cancer by video-assisted thoracoscopic surgery and 630 patients who underwent pulmonary resection via open thoracotomy (as controls) in Cancer Institute & Hospital, Chinese Academy of Medical Sciences between January 2009 and August 2011 were retrospectively reviewed. The risk factors after lobectomy were also analyzed.</p><p><b>RESULTS</b>In the 506 NSCLC patients who received VATS lobectomy, postoperative complications occurred in 13 patients (2.6%) and one patient died of acute respiratory distress syndrome (0.2%). In the 521 patients who received open thoracotomy (OT) lobectomy, postoperative complications occurred in 21 patients (4.0%) and one patient died of pulmonary infection (0.2%). There was no significant difference in the morbidity rate (P > 0.05) and mortality rate (P > 0.05) between the VATS group and OT group. In the 190 patients who received VATS wedge resections, postoperative complications occurred in 3 patients (1.6%). One hundred and nine patients received OT wedge resections. Postoperative complications occurred in 4 patients (3.7%). There were no significant differences for morbidity rate (P = 0.262) between these two groups, and there was no perioperative death in these two groups. Univariate and multivariate analyses demonstrated that age (OR = 1.047, 95%CI: 1.004 - 1.091), history of smoking (OR = 6.374, 95%CI: 2.588 - 15.695) and operation time (OR = 1.418, 95%CI: 1.075 - 1.871) were independent risk factors of postoperative complications.</p><p><b>CONCLUSIONS</b>To compare with the NSCLC patients who should undergo lobectomy or wedge resection via open thoracotomy, a similar short-term outcome can be achieved via VATS approach.</p>

Impact of the reconstruction of the scrotum with third-degree burns on spermatogenic function: experimental study of three different methods / 中华男科学杂志

<p><b>OBJECTIVE</b>To explore the effects of different methods of scrotal reconstruction on the apoptosis of spermatogenic cells and expression of the bcl-2 protein in patients with third-degree scrotal burns.</p><p><b>METHODS</b>Forty male and 24 female 2-month-old Guizhou mini-pigs were used in this study, the former equally randomized to groups I (normal control), II (natural healing), III (skin grafting) and IV (skin flap grafting). Ten months after the establishment of the model of third-degree burns, 6 male pigs from each group were paired with the female pigs and fed for 3 weeks. Then the female pigs were fed for another 4 months, followed by observation of their reproductivity. At 12 months, the bilateral testes were taken from the male pigs for detection of the apoptosis index of spermatogenic cells by TUNEL and determination of the expression of the bcl-2 protein by immunohistochemistry. The data obtained were subjected to single factor analysis of variance.</p><p><b>RESULTS</b>The apoptosis indexes of spermatogenic cells were (7.07 +/- 3.5), (40.34 +/- 4.85), (15.14 +/- 1.36) and (39.29 +/- 5.73)% in groups I , II, III and IV, respectively, significantly higher in groups II , III and IV than in I (P<0.05), with statistically significant differences between group III and groups II and IV (P<0.05) but not between II and IV (P>0.05). The expression rates of the bcl-2 protein were (75.07 +/- 3.74), (54.93 +/- 4.03), (66.85 +/- 3.06) and (53.33 +/- 5.22)% in groups I, II, III, and IV, respectively, remarkably higher in I than in the other three (P<0.05), with significant differences between group III and groups II and IV (P<0.05) but not between II and IV (P>0.05). Pregnancies were found in all the female pigs of group I with 10.0 +/- 1.18 newborns and in 4 of group III with 9.92 +/- 1.31 newborns, but in none of groups II and IV, with significant differences between group I and the other three (P<0.05) as well as between group III and groups II and IV (P<0.05), but not between II and IV (P>0.05).</p><p><b>CONCLUSION</b>All the three methods of reconstruction for the scrotum with third-degree burns can suppress spermatogenic function, increase the apoptosis of spermatogenic cells and decrease the expression of the bcl-2 protein, among which, skin grafting least affects spermatogenic function.</p>

Influence of temperature on spermatogenesis-related genes in spermatogonia cultured in vitro / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the influence of temperature on the expressions of c-kit and PI3K in spermatogonia cultured in vitro at 32 degrees C and 37 degrees C, and provide basic scientific data for the mechanism of spermatogenic impairment due to body temperature (37 degrees C).</p><p><b>METHODS</b>Isolated spermatogenic cells were cultured in vitro at 32 degrees C and 37 degrees C, and their adherence, proliferation and morphologic changes were observed and recorded under the inverted phase contrast microscope. At 8 days, the spermatogonia were separated by Percoll density gradient centrifugation and the differential adhesion method. The expressions of c-kit and PI3K mRNA and proteins in the separated cells were detected by real time polymerase chain reaction and Western blot, respectively. The c-kit gene was sequenced to identify the occurrence of mutations.</p><p><b>RESULTS</b>Adherence, division and proliferation of the cells were observed in both the 32 degrees C and 37 degrees C groups. The expressions of c-kit and PI3K mRNA and proteins in the spermatogonia were significantly higher in the 32 degrees C group than in the 37 degrees C group (P < 0.05). The 32 degrees C group showed no mutation of c-kit in exon 9, 11 and 13; the 37 degrees C group exhibited no mutation in exon 11 and 13, but possible insertion or deletion mutations in exon 9.</p><p><b>CONCLUSION</b>Culturing in vitro at 37 degrees C could inhibit the expression of proliferation- and differentiation-related genes in spermatogenic cells and lead to the mutation of the c-kit gene.</p>

Brucellosis from 2006 to 2010: an analysis of surveillance results at the national surveillance spots / 中国地方病学杂志

ObjectiveTo analyze the surveillance results at the national surveillance spots of brucellosis from 2006 to 2010,to know the epidemic status of national brucellosis,and to provide scientific evidences for evaluating of surveillance quality and formulating of surveillance strategies and measures.Methods The brucellosis surveillance data collected from the Surveillance Reports of Major Infectious Diseases and Vector-Biological Monitoring Report in China from 2006 to 2010,and the National Diseases Surveillance Information Management System were classified and analyzed using two stage clustering statistical analysis.Results According to national routine surveillance,the incidence of brucellosis increased from 1.55 per 100 000 in 2006 to 2.62 per 100 000 in 2010.Reported brucellosis cases were mainly in Inner Mongolia,Shanxi,Heilongjiang and other regions,and reported time was from March to August,which accounting for 79.87％(114 265/143 064).The brucellosis cases were mainly youth,male,farmers and herdsman.In the national surveillance spots of the brucellosis,217 648 cases received epidemic survey,of which 61 905 cases received serological test,the positive rate was 19.66％(12 169/61 905),and 10 318 new cases were found; the highest positive rate of serological test was 41.19％(2757/6694) which was found in Zhalantun of Inner Mongolia; the detection rate of pathogen culture was 2.49％(29/1165); the positive rates of serological test in cattle and sheep were 1.07％(2355/219 352) and 0.93％ (2766/296 176),respectively.The surveillance spots were classified into four grades according to cluster analysis.Conclusions The epidemic continues to rise in human surveillance spots of brucellosis.Although national surveillance spots play an important role in the surveillance of brucellosis,the quality and quantity of surveillance work of national brucellosis surveillance spots should be improved,different management measures should be taken in different surveillance spots according to classified results of the cluster analysis.This study provides some basis to improve the utilization of health resources and the level of Brucella disease prevention and control work.

Identification of biological characteristics of human keloid-derived stem cells / 中华烧伤杂志

<p><b>OBJECTIVE</b>To investigate the biological characteristics of human keloid-derived stem cells (KDSC) in order to further research its role in keloid pathogenesis.</p><p><b>METHODS</b>Human keloid specimens were harvested to isolate and select KDSC by enzyme digestion and subculturing. Primary and (or) the third generation of KDSC were collected for identification of biological characteristics as follows. (1) After addition of mouse anti-human monoclonal fluorescent antibodies (CD29-PE, CD34-PE, CD44-FITC, CD90-FITC, CD45-PerCP), the expression of cell surface antigen phenotype (CD29, CD34, CD44, CD45, CD90) as well as cell cycle was analyzed by flow cytometry. (2) After addition of mouse anti-human cell keratin (CK19) monoclonal antibody and mouse anti-human vimentin monoclonal antibody, the expression level of CK19 and vimentin was respectively determined with immunocytochemical method. RT-PCR was used to detect the expression of Oct4. The multipotent differentiation capacity of the first generation KDSC was observed with osteogenic, chondrogenic and adipogenic nutrient media.</p><p><b>RESULTS</b>After being subcultured, the sizes of cells were similar, and the majority of them were spindle-shaped with disorderly arrangement. The cells highly expressed typical surface markers of mesenchymal stem cells (such as CD29, CD44, and CD90, etc.) with low expression of hematopoietic stem cell surface markers (such as CD34, CD45, etc.). 67.66% of cells were in G0/G1 phase, 26.24% of cells were in G2/M phase, and 6.11% of cells were in S phase. Vimentin was positively expressed in KDSC while CK19 was negatively expressed. The expression of Oct4 was also positive. After being cultured in inducing differentiation media, the cells could differentiate into osteoblasts, chondrocytes, and adipocytes.</p><p><b>CONCLUSIONS</b>Stem cells existing in human keloid, which are similar to mesenchymal stem cells, may play an important role in keloid pathogenesis.</p>

Spermatogenic cell apoptosis and expressions of Bcl-2 and Bax proteins after burying the testis in the inguinal pocket / 中华男科学杂志

<p><b>OBJECTIVE</b>To observe the apoptosis of spermatogenic cells and the expressions of Bcl-2 and Bax proteins after burying the testis in the inguinal pocket, and to investigate their relationship.</p><p><b>METHODS</b>We randomly divided 36 healthy male New Zealand white rabbits into an experimental group (n = 18) and a control group (n = 18). Models were established by burying testes in the inguinal pocket in the experimental group, while the controls were left untreated. At the end of the 8th week after surgery, 6 animals were randomly taken from each group for measurement of the testis surface temperature and testicular biopsy. The apoptosis of spermatogenic cells in the testis tissues was detected by TUNEL assay, and the expressions of Bcl-2 and Bax proteins determined by immunohistochemistry and imaging analysis.</p><p><b>RESULTS</b>At 8 weeks after burying the testis in the inguinal pocket, the testicular surface temperature was significantly higher in the experimental group than in the control ([ 38.02 +/- 0.36] degrees C vs [36.15 +/- 0.64 ] degrees C, P < 0.05), and so was the apoptosis index (AI) of spermatogenic cells ([89.69 +/- 3.76] % vs [7.73 +/- 4.95 ] %, P < 0.05). The expression of the Bax protein in the testis was significantly increased, while that of the Bcl-2 protein remarkably decreased in the experimental group as compared with the control group (P < 0.05). The apoptotic cells were mostly primary spermatocytes and round spermatids.</p><p><b>CONCLUSION</b>Elevated local temperature of the testis buried in the inguinal pocket increases the apoptosis of spermatogenic cells, and the spermatogenic cell apoptosis is highly correlated with the decreased expression of Bcl-2 and increased expression of Bax. The changes in the expressions of Bcl-2 and Bax proteins were a main mechanism behind the temperature elevation-induced apoptosis of spermatogenic cells.</p>

Risk factors on the recurrence of ischemic stroke and the establishment of a Cox' s regression model / 中华流行病学杂志

Objective To investigate the risk factors and establish the Cox' s regression model on the recurrence of ischemic stroke. Methods We retrospectively reviewed consecutive patients with ischemic stroke admitted to the Neurology Department of the Hebei United University Affiliated Hospital between January 1,2008 and December 31,2009. Cases had been followed since the onset of ischemic stroke. The follow-up program was finished in June 30, 2010. Kaplan-Meier methods were used to describe the recurrence rate. Monovariant and multivariate Cox' s proportional hazard regression model were used to analyze the risk factors associated to the episodes of recurrence.And then, a recurrence model was set up. Results During the period of follow-up program, 79 cases were relapsed,with the recurrence rates as 12.75% in one year and 18.87% in two years. Monovariant and multivariate Cox' s proportional hazard regression model showed that the independent risk factors that were associated with the recurrence appeared to be age (X1)(RR=1.025,95% CI: 1.003-1.048),history of hypertension (X2) (RR= 1.976, 95% CI: 1.014-3.851), history of family strokes (X3) (RR=2.647,95%CI: 1.175-5.961), total cholesterol amount (X4) (RR= 1.485,95%CI: 1.214-1.817), ESRS total scores (X5) (RR= 1.327,95%CI: 1.057-1.666) and progression of the disease (X6) (RR= 1.889,95%CI: 1.123-3.178). Personal prognosis index (PI) of the recurrence model was as follows: PI=0.025X1 + 0.681X2+ 0.973X3 + 0.395X4+ 0.283X5 + 0.636X6. The smaller the personal prognosis index was, the lower the recurrence risk appeared, while the bigger the personal prognosis index was, the higher the recurrence risk appeared. Conclusion Age, history of hypertension, total cholesterol amount, total scores of ESRS, together with the disease progression were the independent risk factors associated with the recurrence episodes of ischemic stroke. Both recurrence model and the personal prognosis index equation were successful constructed.

The expression of Cyclin A and p21cip1 in fibroblast of hypertrophic scar / 中华整形外科杂志

<p><b>OBJECTIVE</b>To study the relation of the mRNA and protein expression of Cyclin A and p21cip1 in different stages hypertrophic scar fibroblast (FB) with its cell cycle, so as to provide theoretical evidence for intervention therapy of cell cycle gene being used in hypertrophic scar.</p><p><b>METHODS</b>The hypertrophic scar samples at different stages (the third month, the sixth month, the twelfth month, the twenty-fourth month) in 32 cases and the normal skin samples in 8 cases were used in this study. The expression of Cyclin A, p21cip1 mRNA and protein was detected by quantitative real-time PCR and Western Blot. And at the same time, the flow cytometer was used to detect the fibroblastic cell life cycle.</p><p><b>RESULTS</b>1) The expression of Cyclin A mRNA and protein in the third month group, the sixth month group, the twelfth month group, the twenty-fourth month group were 19.34 +/- 2.41, 0.99 +/- 0.11; 19.30 +/- 1.42, 0.96 +/- 0.09; 10.73 +/- 2.93, 0.66 +/- 0.58; 9.29 +/- 0.97, 0.65 +/- 0.14, respectively. And in corresponding stages, the expression of p21cip1 mRNA and protein were 2.80 +/- 0.69, 0.35 +/- 0.07; 4.95 +/- 1.82, 0.44 +/- 0.07; 9.98 +/- 1.19, 0.56 +/- 0.06; 10.25 +/- 1.46, 0.59 +/- 0.06, respectively. The expression intensity of Cyclin A mRNA and protein was no significantly different between the third month group and the sixth month group (P > 0. 05), but it was higher respectively than that in the twelfth month group, the twenty-fourth month group and normal group (P < 0.05). And the expression intensity was no different between the above three groups (P > 0.05). The expression intensity of P21cip1 mRNA and protein in the third month group was lower than that in the sixth month group, but that in the above two groups was lower respectively than that in the twelfth month group, the twenty-fourth month group and normal group (P < 0.05). And the expression intensity had no difference between the three later stage groups (P > 0.05). 2) Most FB were in S and G2/M stage (cycle) in 3rd month, 6th month group. And most FB were in G0/G1 stage (cycle) in 12th month, 24th month group and normal group.</p><p><b>CONCLUSIONS</b>The expression of mRNA and protein of Cyclin A in hypertrophic scar changes from high level to low level as the hypertrophic scar develops, while the expression of P21cip1 changes from low level to high level. The mRNA and protein expression of Cyclin A and p21cip1 respectively are basically corresponded to different stages of hypertrophic scar. The distribution of cell life cycle of fibroblastic are also corresponded to the expression intensity of Cyclin A and p21cip1 in different stages hypertrophic scar. An early stage intervention to the two gene can be effective to prevent from the genesis and development of hypertrophic scar.</p>

Repairing small wounds around ankle by medial planta island flaps pedicled with anterior tibial artery perforator in front of inner malleolus / 中华整形外科杂志

<p><b>OBJECTIVE</b>To 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.</p><p><b>METHODS</b>From 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.</p><p><b>RESULTS</b>All 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.</p><p><b>CONCLUSIONS</b>The medial planta island flaps pedicled with anterior tibial artery perforator can effectively repair the small wounds around ankle with reliable blood supply.</p>

Apoptosis of spermatogenic cells and expression of HSP70 after scrotal reconstruction with skin flap / 中华男科学杂志

<p><b>OBJECTIVE</b>To explore the temperature change at the testis surface, apoptosis of spermatogenous cells and the expression of the heat shock protein 70 (HSP70) after scrotal reconstruction with the skin flap.</p><p><b>METHODS</b>We included 36 healthy New Zealand white rabbits, 24 males and 12 females, in this study, and equally randomized the males into an experimental and a control group. The scrotal of the experimental rabbits were excised and reconstructed with the hypogastric flap, while the controls were left untreated. At the end of the 8th week after surgery, 6 animals were randomly taken from each of the two groups for measurement of the testis surface temperature and testicular biopsy. The apoptosis of spermatogenous cells in the testis tissues was detected by HE staining, and the expression of HSP70 determined by immunohistochemistry and imaging analysis. The other 6 animals exempt from testicular biopsy in each of the experimental and control groups were mated with the female rabbits, and observed for fertility.</p><p><b>RESULTS</b>At the end of the 8th week after scrotal reconstruction, the testicular surface temperature was (38.1 +/- 0.6) degrees C in the experimental group, significantly higher than (36.0 +/- 0.30) degrees C before surgery (P < 0.05), and the apoptosis index (AI) of the spermatogenous cells was (71.85 +/- 2.7) %, as compared with (7.73 +/- 4.95) % in the control group (P < 0.05). The expression of HSP70 was found mainly in the spermatogenous cells of the experimental group and in the spermatoblasts of the control. A total of 6.0 +/- 1.3 baby rabbits were born in the control group, but none in the experimental group (P < 0.05).</p><p><b>CONCLUSION</b>The testicular surface temperature rises after scrotal reconstruction with the hypogastric flap, which increases the apoptosis of spermatogenic cells and causes infertility. HSP70 is involved in protecting spermatogenic cells from apoptosis after scrotal reconstruction.</p>

Dorsal metatarsal artery distal perforator pedicle link-pattern flaps for wound repairing in the distal feet / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the feasibility of repairing the wounds in the distal feet with dorsal metatarsal artery distal perforator pedicle link-pattern flaps.</p><p><b>METHODS</b>Since January of 2004 to April of 2009, 30 patients with distal wounds in the feet underwent surgical wound repair using by dorsal metatarsal arteries distal perforator pedicle link-pattern flaps. Seventeen patients used the 1, 2 dorsal metatarsal artery distal perforator flaps, and 13 had the 3, 4 dorsal metatarsal artery distal perforator flaps, with the flaps measuring 4 cmx4 cm to 8 cmx7cm and the flap pedicle ranged 3-6 cm in length. The donor sites were repaired with skin grafting.</p><p><b>RESULTS</b>Twenty-nine flaps survived completely and 1 flap showed partial necrosis in the distant part (2.0 cmx1.0 cm). Twenty-one patients were followed up for an average of 11 months, during which the color, texture, and contour of the flaps remained normal without ulcers in the donor sites or the flaps.</p><p><b>CONCLUSION</b>The wounds in the distal feet can be repaired by dorsal metatarsal artery distal perforator pedicle link-pattern flaps, which is a simple, applicable and safe procedure.</p>