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.

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>

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>

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>

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>

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>

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>

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>

Repair of wounds in popliteal fossa scar with perforator flaps of gastrocnemius medial head / 中华烧伤杂志

<p><b>OBJECTIVE</b>To study the effect of perforator flaps of medial head of gastrocnemius muscle on repairing popliteal fossa scar wounds.</p><p><b>METHODS</b>Ten patients with scar in popliteal fossa hospitalized from January 2005 to January 2010 were repaired with perforator flaps of medial head of gastrocnemius muscle. The scar was resulted from burn in 8 patients, and from operation in 2. The duration of the scar was 3 months to 11 years, and area of the scar ranging from 6 cm × 3 cm to 10 cm × 6 cm. Ultrasonic Doppler was used to detect the musculocutaneous perforator vessel of the medial sural artery at the position 10 to 17 cm from the fold of the popliteal fossa and 2 to 5 cm from the posterior midline before surgery. Then flap transplantation surgery was performed. Donor site wounds with width less than 5 cm were sutured directly, and those wider than 5 cm were repaired with skin transplantation.</p><p><b>RESULTS</b>All the flaps survived. Flap size ranged from 7 cm × 5 cm to 12 cm × 7 cm. All patients were followed up for 3 to 30 months, and the flaps were found to have a good appearance. Patients could walk with heavy load without lameness. The function of knee joint of the affected limb was the same as that of the opposite limb. No obvious depression was observed in donor sites.</p><p><b>CONCLUSIONS</b>The perforator flaps of medial head of gastrocnemius muscle can be used to repair the popliteal fossa scar wound with satisfactory result.</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>

Repair of skin defects at forepart of feet with reverse saphenous nerve neurocutaneous flaps / 中华整形外科杂志

<p><b>OBJECTIVE</b>To investigate the effect of reverse saphenous nerve neurocutaneous flaps for skin defects at forepart of feet.</p><p><b>METHODS</b>From January 2004 to October 2008,15 cases of skin defects at forepart of feet were repaired with reverse saphenous nerve neurocutaneous flaps. The flap size ranged from 3.5 cm x 3.0 cm to 8 cm x 5 cm. The wounds at donor site were closed with skin graft.</p><p><b>RESULTS</b>All the flaps survived completely with no ulcer at the donor site. 10 patients were followed up for 1 to approximately 9 months. The skin color and texture were satisfactory. The patients could walk very well.</p><p><b>CONCLUSIONS</b>It is reliable to repair the skin defects at forepart of feet with reverse saphenous nerve neurocutaneous flaps. It is easily performed with less morbidity.</p>

Fibular artery perforator link-pattern flaps at lateral and posterior part of leg / 中华整形外科杂志

<p><b>OBJECTIVE</b>To investigate the feasibility of fibular artery perforator link-pattern flaps at lateral and posterior part of legs.</p><p><b>METHODS</b>From January 2007 to March 2009, 12 cases of with feet and ankle wounds were treat with fibular artery perforator link-pattern flaps at lateral and posterior part of legs. The flap size ranged from 10 cm x 10 cm to 25 cm x 13 cm. The wounds at the donor sites were closed with skin grafts.</p><p><b>RESULTS</b>All the flaps survived completely. The patients were followed up for 1-12 months (median, 6 months). The color, texture and appearance of the flaps were good.</p><p><b>CONCLUSION</b>The link-pattern flap has reliable blood supply and a large area for repairing defects. The sural nerve is reserved, resulting minimal morbidity to donor site.</p>

Anatomic study of the medial pedis composite flaps with saphenous nerve and tendon and its application / 中华整形外科杂志

<p><b>OBJECTIVE</b>To investigate the anatomy of the medial pedis composite flaps with saphenous nerve and tendon and its application in the repair of tendo calcaneus and adjacent defects.</p><p><b>METHODS</b>10 cadavers (20 sides) were observed. The origin, course, size and the distribution of the medial plantar artery were studied. 12 cases with tendo calcaneus and adjacent defects were reconstructed with the medial pedis composite flaps with saphenous nerve. Donor site defects were covered with free skin graft.</p><p><b>RESULTS</b>The medial plantar artery gives off deep branch [diameter (1.5 +/- 0.3) mm] and superficial branch [diameter (1.0 +/- 0.2) mm]. In 18 sides, the deep branches give off the medial branches and lateral branches. While in 2 sides, the superficial branches give off the medial branches and lateral branches with no big branches from the deep branches. There are branches of saphenous nerve and medial dorsal cutaneous nerve in the flap. All the flaps were survived. 8 cases were followed up for one months to one years. Good color, texture and function of the flaps were achieved.</p><p><b>CONCLUSIONS</b>The medial pedis composite flaps with saphenous nerve can repair tendo calcaneus and adjacent defects. It is a easy and safe procedure with reliable anatomy and good results.</p>

Effect of scrotal reconstruction with flap on rabbit generation function / 中华整形外科杂志

<p><b>OBJECTIVE</b>To explore the apoptosis and the express of proliferating cell nuclear antigen (PCNA) in spermatogenic cells, and study generation function of the rabbit after scrotal reconstruction with flaps.</p><p><b>METHODS</b>The 48 New Zealand white rabbits were randomly divided into three groups as experimental group (the scrotum reconstructed with flaps, n = 48), the control group (the sham operated group, n = 24) and the blank group (n = 18). The apoptosis and the expression of PCNA in the spermatogenic cells were detected with TUNEL and the immunohistochemistry from the 3rd to the 8th week after operation. 8 weeks later, 12 animals in each group were fed respectively with one female rabbit to observe the procreation.</p><p><b>RESULTS</b>The apoptotic index of the spermatogenic cells in blank group was 7.73 +/- 4.95. 3 weeks after operation, the apoptotic index of spermatogenic cells was 22.59 +/- 3.04 in the experimental group, and 21.13 +/- 1.68 in control group, showing no significant difference between the two groups (P > 0.05). 8 weeks after operation, the apoptotic index of spermatogenic cells was 71.85 +/- 2.69 in the experimental group, and 13.64 +/- 2.09 in control group, show a significant difference between them (P < 0.05). The apoptotic index in experimental group increased gradually from the 3rd to 8th week after scrotal reconstruction , which was markedly higher than that in the blank group (P < 0.05). The apoptotic index in control group was higher than that in the blank group at the 3rd week (P < 0.05), but not at the 8th week (P > 0.05). The proliferation index of spermatogenic cells was 9.32 +/- 9.30 and 12.52 +/- 3.87 in experimental group at the 3rd and 4th week, respectively, which was significantly lower than that in blank group (43.07 +/- 2.25) and control group (45.69 +/- 4.98) at the 3rd week (P < 0.05). The proliferation index of spermatogenic cells was 46.98 +/- 18.92 and 49.53 +/- 9.79 in experimental group at the 7th and 8th week, respectively, 39.90 +/- 5.10 in control group at the 8th week, showing no difference between the two groups (P > 0.05). The proliferation index of spermatogenic cells in control group at the 3rd and 8th week was not different from that in the blank group (P > 0.05). The female pairing rabbits in the blank and control group were all pregnant, and the average childbirths were 6.0 +/- 1.28 and 5.92 +/- 1.31 respectively, with no difference between the two groups (P > 0.05). All the female pairing rabbits in the experimental group were not pregnant, showing a significant difference from those in the blank and control group (P < 0.05).</p><p><b>CONCLUSIONS</b>The rabbit generation functional disturbance after scrotal reconstruction with flaps is due to the excessive apoptosis of spermatogenic cell. The spermatogenic cell proliferation is affected only in the early postoperative period, but can recover later.</p>