Sijunzi Decoction Inhibits Stemness by Suppressing ß-Catenin Transcriptional Activity in Gastric Cancer Cells.

OBJECTIVE: To investigate a previously uncharacterized function of Sijunzi Decoction (SJZD) in inhibition of gastric cancer stem cells (GCSCs). METHODS: MKN74 and MKN45, two CD44 positive gastric cancer cell lines with stem cell properties were used. The cells were divided into 2 groups. Treatment group was treated with SJZD (1-5 mg/mL) for indicated time (48 h-14 days). The control group was treated with equal volume of phosphate buffered saline. Cell Counting Assay Kit-8 were used to measure cell viability. Spheroid colony formation and GCSCs marker expression were performed to determine GCSCs stemness. Cell fractionation and chromatin immunoprecipitation assays were used to assess the distribution and DNA-binding activity of ß-catenin after SJZD treatment, respectively. RESULTS: SJZD treatment repressed cell growth and induced apoptosis in MKN74 and MKN45 cell lines (P<0.05). Moreover, SJZD dramatically inhibited formation of spheroid colony and expression of GCSC markers in GC cells (P<0.05). Mechanistically, SJZD reduced nuclear accumulation and DNA binding activity of ß-catenin (P<0.05), the key regulator for maintaining CSC stemness. CONCLUSION: SJZD inhibits GCSCs by attenuating the transcriptional activity of ß-catenin.

Zeolite-based hemostat QuikClot releases calcium into blood and promotes blood coagulation in vitro.

AIM: To examine the changes in electrolyte concentrations after addition of zeolite-based hemostat QuikClot in blood and the effects of zeolite on blood coagulation in vitro. METHODS: Fresh blood was taken from healthy adult volunteers and sheep, and the electrolyte concentrations in blood were measured using a blood electrolyte analyzer. Zeolite Saline Solution (ZSS) was prepared by addition of 2 g zeolite to 0.9% NaCl solution (4, 8, or 16 mL). The electrolytes in ZSS were measured using inductively coupled plasma atomic emission spectroscopy. The prothrombin time (PT) and activated partial thromboplastin time (APTT) of blood were measured using the test tube method. The activated clotting time (ACT) and clotting rate (CR) of blood were measured with Sonoclot Coagulation and Platelet Function Analyzer. RESULTS: Addition of zeolite (50 and 100 mg) in 2 mL human blood significantly increased Ca(2+) concentration, while Na(+) and K(+) concentrations were significantly decreased. Addition of zeolite (50 and 100 mg) in 0.9% NaCl solution (2 mL) caused similar changes in Ca(2+) and Na(+) concentrations. Si(4+) (0.2434 g/L) and Al(3+) (0.2575 g/L) were detected in ZSS (2 g/8 mL). Addition of ZSS in sheep blood shortened APTT in a concentration dependent manner, without changing PT. ZSS or aqueous solution of CaCl2 that contained Ca(2+) concentration identical to that of ZSS significantly shortened ACT in human blood without significantly changing CR, and the effect of ZSS on ACT was not significantly different from that of CaCl2. CONCLUSION: Zeolite releases Ca(2+) into blood, thus accelerating the intrinsic pathway of blood coagulation and shortening the clot formation time.

Epidemiological studies of electrical injuries in Shaanxi province of China: a retrospective report of 383 cases.

A statistical survey was conducted at the Burn Unit of the Tangdu Hospital, Shaanxi, China, during the 10-year period from January 2000 to December 2009. In this retrospective study, 383 patients who admitted to our burn unit because of electrical trauma were included. Data including the patient's general condition, clinical presentation, complications and operation times was collected retrospectively and analyzed with epidemiological methods. Subjects in our collective were predominantly male (90.3%, n=346/383) and were composed by those who injured in work-related incidents (78.3%, n=300/383), rural individuals (58.2%, n=223/383) and students (9.4%, n=36/383). High voltage was directly correlated to severity clinical complications, and amputation. The percentage of myocardial impairment was 79.3% (n=92/116) among patients who suffered with electrical current through heart tissue. Along with the more developed east area of China, electrical injuries are becoming a growing concern of the developing West part in China as well. Electrical injuries induce serious tissue damage, need long hospital stay, and result in high rate of permanent disability and economic hardship for the afflicted families. A competent prevention program needs to be developed to address this problem.

Autologous nerve implantation into denervated monkey skin promotes regeneration of Meissner's corpuscle.

BACKGROUND: The aim of this study was to observe the effects of autologous nerve implantation into the denervated finger flap on the regression and regeneration of sensory nerve endings and Meissner's corpuscles. MATERIAL/METHODS: Bilateral nerves of fingers were separated: one was removed and the other was implanted into the denervated finger in the implantation group. In the non-implantation group, both nerves were removed. The ventral skin of fingers was collected for immunohistochemistry and electron microscopy 3, 6, 9 and 12 months after surgery. RESULTS: The nerve endings in the Meissner's corpuscles began to degenerate 3 months after denervation. The elementary structure of Meissner's corpuscles was not significantly altered. Nerve fibers were present around the Meissner's corpuscles, accompanied by growing into its inward. The axons in the denervated nerve disappeared and the Meissner's corpuscles began to atrophy at month 6. More regenerated nerve fibers were observed after nerve implantation, including intensive and thick fibers, accompanied by reinnervation of Meissner's corpuscles. More nerve fibers and a higher proportion of myelinated nerve fibers were noted at month 9 in the implantation group, and the reinnervation was present in the majority of Meissner's corpuscles. Naive myelinated nerve fibers appeared at the caudal end of Meissner's corpuscles. The nerve fibers in the Meissner's corpuscles increased to the normal level at 12 months after nerve implantation. CONCLUSIONS: The implanted nerve regenerated a large amount of free nerve endings, which helped to regenerate simple Meissner's corpuscles via governing previously degenerated corpuscles.

Electrical injury alters ion channel expression levels and electrophysiological properties in rabbit dorsal root ganglia neurons.

The electrophysiological and morphological changes of nerve fibers induced by electrical injury have been widely addressed. However, the changes of ion channels in neurons after electrical shocks have not been systematically investigated yet. In this study, the sciatic nerves of rabbit were injured by 50 V 50 Hz, 110 V 50 Hz, and 220 V 50 Hz alternating current, respectively. One week later, the expression levels and electrophysiological changes of voltage-gated potassium (Kv) and sodium (Nav) channels in dorsal root ganglia (DRG) neurons were evaluated by RT-PCR, immunofluorescence staining and patch clamp technique. The Nav1.1 expression was decreased by 50V injury. The Kv1.2, Kv1.4, Nav1.1 and Nav1.7 expression levels and Kv current densities were reduced after 110 V injury. Under the 220 V injury circumstance, Kv1.2, Nav1.1, Nav1.7 expression levels, Kv current densities and TTX-R Na(+) current densities were significantly decreased, while TTX-S Na(+) current densities increased. These findings suggest that the expression levels, subunit compositions, and electrophysiological properties of Kv and Nav channels are altered after electrical injury, and the severity of injury gets worse as injury voltage increases.

[Observation on the fluid resuscitation and the change in blood sodium of severely burned patients in the early stage].

OBJECTIVE: To study the necessary amount of fluid consisting of electrolyte and colloid, the ratio of electrolyte and colloid used, and the change of blood sodium during early resuscitation in severely burned patients. METHODS: Sixty-seven patients with total burn surface area (TBSA) equal to or over 70% and full-thickness area equal to or over 50%TBSA, hospitalized from March 2004 to March 2009, were resuscitated with fluid. The infusion amount of electrolyte, colloid, and water, and urinary output of patients at post injury hour (PIH) 24, 48, and 72 were analyzed retrospectively. The variation in blood sodium and fluid infusion at different time points was recorded. Data were processed with SPSS 13.0 software. RESULTS: Among the 67 patients, hyponatremia occurred in 9 cases, hypernatremia occurred in 5 cases, and 53 patients had normal blood sodium level. The urinary output of patients within PIH 72 was above 70 mL/h. K value was calculated through the formula: actual total infusion amount of electrolyte and colloid (mL) = burn area (%TBSA) x body weight (kg) x K. In the first 24 PIH, K value was about 1.7, and the ratio of electrolyte and colloid was 1.4. In the second 24 PIH, K value was about 1.3 with electrolyte and colloid ratio 1.6. K value in the third 24 PIH was about 0.9 with electrolyte and colloid ratio 2.0. CONCLUSIONS: The actual amount of resuscitation fluid is slightly larger than that calculated from traditional formula during the early stage in severely burned patients. The amount of electrolytes and the proportion of electrolyte and colloid will influence blood sodium level of patients.

[Therapeutic effect of vacuum-assisted closure technology on infected explosion wound of pig].

OBJECTIVE: To investigate the effect of vacuum-assisted closure (V. A. C.) technology in the treatment of infected wound of skin and soft tissue as a result of explosion injury in pig. METHODS: Sixteen explosion wounds were established by electric detonators on the shoulders and hips on both sides of 4 small white domestic pigs ,and they were divided into A group [(without treatment and infection occurred on 1-2 post burn day (PBD), then treated with vaseline gauze on 3 (PBD)], and B group (with the same treatment as in A group, except for treatment of vacuum assisted closure (V. A. C) with pressure of - 15 kPa after 3 PBD). The data of wound depth, wound area, wound healing time were collected and analyzed at 3 PAD and 1, 3, 6, 9, 14, 19, 24 days after treatment. Specimens from wounds were collected for histopathology observation, including also cell proliferation index, the number of vascular endothelial cells, the activity of myeloperoxidase (MPO) and the number of bacteria. RESULTS: Compared with those in A group on land 3 days after treatment, wound area, wound depth were not enlarged or deepened in B group, while the number of inflammatory cells, vascular endothelial cells, proliferative cells were increased, the activity of MPO was enhanced and the number of bacteria was decreased. There were obvious differences between two groups in following indices: wound area ,wound depth, the number of vascular endothelial cells and bacteria during 1 to 19 days after treatment (P < 0.01)), the number of cell proliferation from 1 - 9 days after treatment (P < 0.01)), and the activity of MPO on 3, 6 days after treatment (P < 0. 01). The wound healing time was (32.8 +/- 1.6) d in A group, which was longer than that in B group (25.8 +/- 1.0 d, P < 0.01). CONCLUSION: Compared with conventional dressing change, V. A. C can decrease bacteria load, lessen secondary necrosis, prompt the inflammatory response, accelerate the formation of granulation tissue, shorten wound healing time in infectious wound of porcine skin and soft tissue resulted from explosion injury.

[The influence of vacuum-assisted drainage on the growth of capillaries in the wound produced by explosion in pig].

OBJECTIVE: To investigate the influence of vacuum-assisted closure (VAC) technique on the growth of capillaries in the wound of the pig produced by explosion. METHODS: Four small white pigs were inflicted with 16 explosion wounds [(7.3 +/- 1.0) cm2 in area] on both sides of the buttocks, shoulders and hips by detonation of a specific type of explosive, and the wounds were randomly divided into 2 groups, i. e, control (C, with conventional treatment from 2 post-injury day (PID) on and treatment (T, with VAC treatment after debridement from 2 PID on) groups, with 8 wounds in each group. Wound tissues of 2mm x 2mm x 2mm in size were harvested for pathological examination before treatment and on 1 and 3 post-treatment day (PTU). The differentiation of adventitial cells were examined with light microscope, and the pixel value of desmin positive particles and the luminal area of newly formed capillaries were assessed with Image C software. RESULTS: Most of vessels in the wound of both groups were in elliptic shape when observed in longitudinal section. In C group, few newly formed capillaries vessels with lack of pericytes were observed before treatment and on 1, 3 PTD, then the number began to increase on 6 PTD. In T group, the number of newly formed capillaries with pericytes was increased on 1 PTD, and it continued to increase thereafter. The pixel values of desmin positive particles in C group on 1, 3, and 6 PTD were (91 +/- 54), (199 +/- 85), and (1552 +/- 298), respectively, which were obviously higher than those in T group [(2569 +/- 330), (3984 +/- 377), (9611 +/- 960), P < 0.01]. The area of vessel lumen in C group was (59 +/- 36), (250 +/- 70), and (938 +/- 287) microm2, respectively on 1, 3, and 6 PTD, which was also smaller than those in T group [(818 +/- 234), (4518 +/- 1080), and (9058 +/- 1656) microm2, P < 0.01]. CONCLUSION: Compared with conventional therapy, VAC can not only accelerate the formation of new capillaries, but also enhance the differentiation of pericytes and the process of enwrapping them around the vessels, and increase the luminal area of newly formed capillaries.

[The changes in blood flow in sciatic nerve after electrical injury in rabbit].

OBJECTIVE: To study the changes in sciatic nerve blood flow and the expression of collagen type I after electric injury of rabbit nerve with different voltages. METHODS: Thirty-six healty rabbits were randomized into 3 groups before receiving injury with electricity in voltages, i.e. 50 v, 75 v, and 100 v groups. The changes in blood flow of sciatic nerve were observed with Laser Doppler Flowmeter immediately after injury and 1, 4, 8 weeks after injury. The changes in the expression of collagen type I was observed by immunohistochemical method, and the positive expression rate was calculated. RESULTS: The sciatic nerve blood flow increased in all groups immediately after electric injury. In the 75 v and 100v groups, the nerve blood flow [(53 +/- 3 ), (48 +/- 5) PU] was obviously lower than that of normal value [(62 +/- 4) PU, P < 0.05]. There was little collagen type I deposition in 50 v group, while brown collagenous fibers in epineurium and perineurium were observed in 75 v and 100v groups 4 and 8 weeks after injury. The expression of collagen type I in all groups were obviously higher than that of normal value, and that in 75v and 100 v groups were higher than that in 50 v group at bachl time-point (P < 0.01). CONCLUSION: The restoration of sciatic nerve blood flow is postponed following by the injury with increase of the electrical voltage. The collagen deposition after electrical injury may be one of the reasons for nerve blood flow decrease.

[Effect of vacuum-assisted closure on the expression of urokinase-type plasminogen activator and urokinase-type plasminogen activator receptor in acute and chronic wounds healing].

OBJECTIVE: To study the effect of vacuum-assisted closure (V.A.C) on the expression of Urokinase-type plasminogen activator (uPA) and Urokinase-type plasminogen activator receptor(uPAR) protein in margin tissue of pigs with acute wounds and patients with chronic wounds. METHODS: Acute wounds were created on the two side of five male pigs' back, the experiment wounds on one side received V. A. C treatment and the control side received traditional treatment. Punch biopsies were taken from margin tissue of the wounds in 0, 1, 3, 6, 9, 12, 18, 25 days after the V.A.C treatment. The uPA and uPAR positive cells were stained with immunohistochemical technique . Six human chronic wounds were also treated with the V. A. C treatment, and the samples of extravasate from those wounds were collected in 0, 1, 3, 5, 7 days after the treatment, and the levels of uPA and uPAR expression were examined by enzyme-linked immunosorbent assay (ELISA). RESULTS: The expression of uPA and uPAR protein in margin tissues of pigs with acute wounds increased and peaked in 3 days after the treatment with V. A. C, then it presented rapidly downtrend, but the expression and staining in the experiment group were obviously higher than that of the control group. In the six chronic wounds, the high level expression of uPA and uPAR protein was decreased after the treatment with V. A. C. CONCLUSION: The V. A. C may increase the expression of uPA and uPAR protein in acute wound keratinocytes and decrease the high expression of uPA and uPAR in chronic wounds.

[Repair of peripheral nerve defect by xenogeneic acellular nerve basal lamina scaffolds].

OBJECTIVE: To explore the possibility to bridge peripheral nerve defects by xenogeneic acellular nerve basal lamina scaffolds. METHODS: Thirty SD rats were randomly divided into 5 groups; in each group, the left sciatic nerves were bridged respectively by predegenerated or fresh xenogeneic acellular nerve basal lamina scaffolds, autogenous nerve grafting, fresh xenogeneic nerve grafting or without bridging. Two kinds of acellular nerve basal lamina scaffolds, extracted by 3% Triton X-100 and 4% deoxycholate sodium from either fresh rabbit tibial nerves or predegenerated ones for 2 weeks, were transplanted to bridge 15 mm rat sciatic nerve gaps. Six months after the grafting, the recovery of function was evaluated by gait analysis, pinch test, morphological and morphometric analysis. RESULTS: The sciatic nerve function indexes (SFI) were -30.7% +/- 6.8% in rats treated with xenogeneic acellular nerve, -36.2% +/- 9.7% with xenogeneic predegenerated acellular nerve, and -33.9% +/- 11.3% with autograft respectively (P > 0.05). The number of regenerative myelinated axons, diameter of myelinated fibers and thickness of myelin sheath in acellular xenograft were satisfactory when compared with that in autograft. Regenerated microfascicles distributed in the center of degenerated and acellular nerve group. The regenerated nerve fibers had normal morphological and structural characters under transmission electron microscope. The number and diameter of myelinated fibers in degenerated accellular nerve group was similar to that of autograft group (P > 0.05). Whereas the thickness of myelin sheath in degenerated accellular nerve group was significantly less than that of autograft group (P < 0.05). CONCLUSION: The above results indicate that xenogeneic acellular nerve basal lamina scaffolds extracted by chemical procedure can be successfully used to repair nerve defects without any immunosuppressants.