[Study on the correlative factors of the pain under the second metatarsal in patients with hallux valgus].

OBJECTIVE: To analyze the correlation between the hallux valgus angle and arch X-ray parameters change, plantar pressure by measuring the X-ray indexes and foot-pressure indexes in patients with hallux valgus, as well as to study the main cause factors for the pain under the second metatarsal head. METHODS: A retrospective study of 254 patients(477 feet) treated in Wangjing Hospital from January 2012 to June 2013 was performed. The pain under the second metatarsal head and age distribution were recorded. All the patients were divided into two groups according to the second plantar bone pain: pain group and no pain group. The following indexes were measured and compared: HAVA(hallux abductor valgus angle), IMA1-2(the inter-metatarsal angle between the first and second metatarsals), IMA1-5(the inter-metatarsal angle between the first and fifth metatarsals), TAOTMLA (top angle of the medial longitudinal arch), AAOTMLA (anterior angle of the medial longitudinal arch), SMRL (the second metatarsal relative length than the first), and the plantar pressure indicators including TPUM (touch the ground time percentage under the second metatarsal head), PPUM (peak pressure under the second metatarsal head), and IUM(impulse under the second metatarsal head). All the factors were evaluated by Logistic regression analysis. RESULTS: Spearman rank correlation test showed that there were statistical significance in correlation between HAVA and IMA1-2, IMA1-5, TAOTMLA, AAOTMLA, TPUM(P<0.05, r=0.647, 0.553, 0.127, -0.165, 0.158). Factors including the HAVA, SMRL and the TPUM were the risks for the pain under the second metatarsal head in patients with hallux valgus(P<0.05, ORj=1.030, 1.069, 1.060). CONCLUSIONS: Increase of the hallux valgus angle causes the collapse of hallux valgus arch, extending the weight bearing time of the second metatarsal and increasing the possibility of the pain under the second metatarsal.

Clinical value of polymerase chain reaction in the diagnosis of joint tuberculosis by detecting the DNA of Mycobacterium tuberculosis.

OBJECTIVE: To assess the clinical value of polymerase chain reaction (PCR) in the diagnosis and differential diagnosis of joint tuberculosis (TB). METHODS: PCR was used blindly to detect the DNA of Mycobacterium tuberculosis (M.TB) in five specimens of M.TB, 5 of BCG, and 10 of other bacteria. Then, M. TB in 98 samples from patients with joint TB and 100 samples from patients with non-tubercular joint disorders were detected by PCR, acid-fast staining and culture,. The sensitivity, specificity, accuracy, positive predictive value, and negative predictive value of PCR were calculated. The χ2 test was used for statistical analysis of the frequency of various factors. At the same time, some problems with PCR were also systematically analyzed. RESULTS: (1) In the "standard samples", both M. TB and BCG showed positive while other bacteria were negative. (2) In 98 cases from patients with joint TB, 81 were positive by PCR, 6 by acid-fast staining, and 17 by culture. In 100 cases from patients with non-tuberculous joint disorders, 9 were positive by PCR, and none by either acid-fast staining or culture. Sensitivity, specificity, accuracy, positive and negative predictive value of PCR were 82.65% (81/98), 91.00% (91/100), 86.87% (172/198), 90.00% (81/90) and 84.26% (91/108), respectively. (3) The positive rates for PCR, acid-fast staining and culture in detection of M. TB were 82.65% (81/98), 6.12% (6/98), and 17.34% (17/98), respectively. There were statistically significant differences between the three methods (P < 0.001). (4) The process of PCR is automatic, and can be completed within 3 to 6 hours, whereas 4 to 8 weeks are required for the conventional culture of M. TB. CONCLUSION: PCR is a sensitive, specific, rapid, simple and minimally invasive method for detection of M. TB in samples from joint TB, and can play an important role in early and rapid diagnosis and differential diagnosis of joint TB. But it also has some limitations, such as false positivity and false negativity.

[Effects on VEGF and VEGF mRNA expression of the tissues of periosteum in three-period treatment of TCM for the fractures in rabbit].

OBJECTIVE: To observe the effects of the three-period treatment theory of bone fracture in TCM (Traditional Chinese Medicine) on VEGF and VEGF mRNA expression in the issues of outer periosteum, endosteum and bone marrow of rabbits, and to explore the rationality of phasing method in TCM in treating fracture. METHODS: 3 mm bone defection were made at lower one third part of both radius in 140 male healthy rabbits. The rabbits were randomly divided into four groups, including three-period treatment group (TTG), one-period treatment group(OTG), positive medicine treatment group(PTG) and model control group (MCG). The rabbits in TG were treated with three-period treatment, rabbits in OTG were treated with one-period treatment, rabbits in PTG were fed by Guzhe-Cuoshangsan (a Chinese patent medicine which was used to treat bone fracture), rabbits in model control group were given no prescription or drug but distilled water as same dose as that of other groups. At day 3, 6, 9, 14, 28, 42 and 56, five rabbits from every group were randomly selected and were killed by aeroembolism. The left radiuses were taken out as the research object. Immunohistochemistry stain and in situ hybridization stain were performed to examinate the VEGF and VEGF mRNA expression in the outer periosteum, endosteum and bone marrow. RESULTS: The VEGF and VEGF mRNA expression of all TCM treatment groups were enhanced in the outer periosteum, endosteum and bone marrow at different time points in fracture healing. The VEGF and VEGF mRNA expression in the three tissues of TTG had the tendency of higher than that of the other groups at the most time points after operation. CONCLUSION: Treating fracture in stages has more predominant effect on the expression of VEGF and VEGF mRNA in the outer periosteum, endosteum and bone marrow than that of treating fracture with single prescription or drug.

Long term efficacy of minimal incision osteotomy for hallux abducto valgus.

OBJECTIVE: To observe and evaluate the long-term results of minimal incision osteotomy for hallux abducto valgus. METHODS: From February 1995 to May 1999, 372 cases (705 feet) with hallux abducto valgus were treated with minimal incision osteotomy. Seventy-nine patients (150 feet) were followed up for more than five years (mean 7.5; range, 5.3-13.2 years) after surgery. The preoperative and postoperative hallux abducto valgus angles (HVA), intermetatarsal angles (IMA), tibial sesamoid position (TSP), American Orthopaedic Foot And Ankle Society (AOFAS) score, range of movement of the first metatarsophalangeal joint and lateral metatarsalgia were observed, measured and evaluated. RESULTS: Based on clinic curative effect evaluation criterion, 56 feet (37.3%) were excellent, 88 feet (58.7%) good, 6 feet (4.0%) fair. The postoperative mean AOFAS score was 84.20 ± 4.32 points. The mean HVA decreased from 33.28° to 12.31° and the mean IMA1-2 from 11.75° to 6.80°. The TSP was corrected from an average preoperative grade of 4.29 to a grade of 3.07 by final follow-up. There was no nonunion or delayed union, no avascular necrosis, no infection, and no hallux varus. Numbness in the big toe was found in 4 feet (2.7%). The range of motion of the first metatarsophalangeal joint decreased from 70.20° to 69.53°. Of 97 feet (64.7%) with pre-operative 2-5 metatarsalgia, this had disappeared in 35 feet, improved in 54 feet and was aggravated in 8 feet postoperatively. CONCLUSION: Minimal incision osteotomy is a simple and reliable technique for treating hallux abducto valgus with minimal complications.

[Case control study of hook needle knife for the treatment of stenosing tenovaginitis of flexor digitorum].

OBJECTIVE: To investigate the effect and safety of the hook needle knife for the treatment of stenosing tenovaginitis of flexor digitorum. METHODS: From September 2007 to September 2008, 60 outpatients with stenosing tenovaginitis of flexor digitorum were randomized divided into the treatment group and the control group, 30 cases in each group. Among the patients, 44 patients were female and 16 patients were male, aged from 34 to 69 years, averaged 56 years, the duration of disease ranged from 1 month to 1 year, averaged 3 months. All the patients were treated with hook needle knife and local-blocking respectively. The patients were followed up for 6 months, and the relief of moving-pain, tender-pain, stretching-pain and resist-ing--pain were observed respectively. All the patients were evaluated by the symptoms with numerical rating scale. RESULTS: The relief of moving-pain, tender-pain, stretching-pain and resisting-pain in the treatment group were significantly better than those of the control group; and the therapeutic effects of treatment group were better than those of the control group. CONCLUSION: The method for treating stenosing tenovaginitis of flexor digitorum with hook needle knife has advantages of definite effects, micro-invasion and safety.

[Comparison study on polymerase chain reaction (PCR) and standard culture technique in detecting mycobacterium tuberculosis to diagnose of joint tuberculosis].

OBJECTIVE: To study the role of PCR technique in detection of mycobacterium tuberculosis in the samples from joint tuberculosis, and to evaluate the clinical value of PCR in diagnosis of joint tuberculosis. METHODS: From June 1993 to August 2001, PCR was used to detect DNA of mycobacterium tuberculosis, and the standard culture was applied to detect mycobacterium tuberculosis. Mycobacterium tuberculosis were respectively blindly by the two techniques in the samples obtained from 95 patients with joint tuberculosis (55 males and 40 females, the age ranging from 2 to 75 years, with an average of 34 years). The positive rate of mycobacterium tuberculosis detection was calculated. RESULTS: In the detection of mycobacterium tuberculosis, positive rate was 82% (78/95) in PCR technique, and 16% (15/95) in standard culture technique. There were statistical differences between the two groups (chi2=67, P<0.001). The whole process of PCR amplification was automatic and could be finished within several hours, and the detecting time was considerably shorter. CONCLUSION: PCR technique is a rapid, simple, sensitive and specific method for detection of mycobacterium tuberculosis in the samples of joint tuberculosis, showing more marked advantages than the standard culture technique. It is valuable in the early rapid diagnosis and differential diagnosis of joint tuberculosis.

[Swanson double-stem silicone implant arthroplasty in the treatment of Freiberg disease].

OBJECTIVE: To observe the short-term results of Swanson double-stem silicone implant arthroplasty in the treatment of late stage Freiberg disease. METHODS: From July 2006 to December 2007, Swanson double-stem silicone implant arthroplasty was performed on 13 patients, 11 male (1 foot) and 12 females (17 feet), suffering from late stage Freiberg diseases. All the cases were the second metatarsophalangeal joints. According to the Smillie classification, 11 feet present with grade 4 osteonecrosis, 7 feet with grade 5. The AOFAS scoring system was used for clinical assessment. All the patients were followed up for an average of 11.3 months (3 to 17 months). RESULTS: The mean preoperative and postoperative AOFAS scores were (50.06 +/- 9.59) and (77.50 +/- 4.99), respectively (P<0.05). The complaint of pain with joint motion was decreased in all patients. The postoperative passive range of motion of joints improved significantly. CONCLUSION: Swanson double-stem silicone implant arthroplasty yields satisfied short-term results in late stage Freiberg disease, which is effective to improve range of motion and relieve pain of joint.

[Treatment of capsulitis of little toe with minimal incisional osteotomy and manipulations].

OBJECTIVE: To study therapeutic effects of capsulitis of little toe treated with minimal incisional osteotomy and manipulations procedures. METHODS: From 2003.7 to 2008.6, 90 patients (160 feet) with capsulitis of little toe including 3 male (5 foot) and 87 female (155 foot) ranging in age from 17 to 76 years(average of 49.2 years).The average medical history was 10.1 years ranging from 1 to 32 years. All patients with capsulitis of little toe were treated with minimal incisional osteotomy and manipulations procedures adopting the modified Coughlin standard to evaluation. RESULTS: All patients were followed up, the duration of follow-up raunged from 3 to 36 months with averagement of 15.7 months. According to the modified Coughlin standard, 80 cases obtained an excellent result, 8 good, 2 fair and 0 poor, the effective rate was 97.8%. However,two patients were found delayed union after operation, the distal fragments healed following the time of fixation were delayed. CONCLUSION: The treatment of capsulitis of little toe with minimal incisional osteotomy and manipulations procedures is easy to operate and its therapeutic effect is convincing.

[Suppression of murine EAE by triptolide is related to downregulation of INF-gamma and upregulation of IL-10 secretion in splenic lymphocytes].

AIM: To explore the therapeutic effectivity and the possible mechanism of triptolide (Tri) on experimental autoimmune encephalomyelitis (EAE). METHODS: All female C57BL/6 mice were randomly divided into EAE group (28), Tri treated group (20) and adjuvant group (18). Mice in EAE and treated groups were immunized with myelin oligodendrocyte glycoprotein peptides 35-55 (MOG(35-55);), adjuvant group was injected at the same time, but instead of MOG(35-55); with normal saline. Tri was intraperitoneally injected in the dosage of 100 microg/(kg.d) in treated group on day 5 post-immunization (p.i.), and mice in EAE and adjuvant group injected with normal saline as control. The clinical feature and pathological changes were observed and the splenic lymphocytes were prepared on days 18-20 p.i. The cell cultures were divided into the control group (only 200 microL of cell suspension) and the experimental group (cell suspension in the presence of 10 mg/L MOG(35-55);). Then all of them were inoculated in 96-well flat-bottom plates under 37 degrees Celsius, 50 mL/L CO(2);. After 48 h, the proliferation assay was determined by MTT, and the supernatants were harvested for the detection of INF-gamma, IL-17, IL-10 and IL-4 by ELISA. RESULTS: Tri treatment showed an significantly protective action on EAE. After the intervention of Tri, the levels of IL-10 were increased (P<0.05), but the secretion of INF-gamma and proliferation response of splenic lymphocytes induced by MOG(35-55); were statistically significantly inhibited(P<0.05 and P<0.01, respectively). There were no influences on the amount of IL-17 and IL-4 by Tri. CONCLUSION: Tri is an effective drug in suppressing murine EAE. This suppression is supposed to be related to downregulation of INF-gamma and upregulation of IL-10 secretion in splenic lymphocytes.

[Observation on pathological changes of the medial aspect of the first metatarsal head of hallux valgus].

OBJECTIVE: To observe pathological characteristic of bone and soft tissue of the medial aspect of the first metatarsal head in order to investigate the pathogenesis of hallux valgus. METHODS: One hundred and twenty-three patients with hallux valgus (235 feet) were obtained from August 2003 to December 2004 in our department. The bone and soft tissue of the medial aspect of the first metatarsal head were taken out from the incision. The pathological conditions of specimen were observed by HE stain. RESULTS: The pathological changes included: degeneration and desmoplasia around the attachment point of tendon and joint capsule, reactive hyperplasia of periost, decreasing of bone trabecula, cartilaginification of periost, cortical bone and tendon insertion, bone formation and osteoclasts response. CONCLUSION: The pathological changes of the medial aspect of the first metatarsal head of hallux valgus presented extensive chronic inflammation.

Tissue engineered cartilage from hTGF beta2 transduced human adipose derived stem cells seeded in PLGA/alginate compound in vitro and in vivo.

Chondrogenic potential of human adipose derived stem cells (hASCs) makes them a possible source of seeding cells for cartilage tissue engineering. We aim to examine the chondrogenic differentiation of human transforming growth factor beta2 (hTGF beta2) transduced hASCs seeded in three-dimensional scaffold in vitro and in vivo. In this study, hASCs were isolated from human subcutaneous adipose tissue and transduced with a replication deficient adenovirus carrying hTGF beta2 (Ad5-hTGF beta2), and then the transduced cells were seeded and cultured in PLGA/alginate compounds. RT-PCR analysis revealed that Ad5-hTGF beta2 transduced hASCs produced aggrecan and collagen type II after 7-day induction in vitro and continued throughout the culture period; this was also demonstrated by the positive staining of Alcian blue and immunohistochemistry for collagen type II. For in vivo study, Ad5-hTGF beta2 transduced hASCs seeded in PLGA/alginate compounds were implanted in subcutaneous pockets of nude mice; after 12 weeks, the implants were harvested and examined by haematoxylin and eosin staining, AB-PAS staining, and immunohistochemical analysis, and the results demonstrated the formation of cartilage tissue. As a control, all these were not observed in the constructs with Ad5-EGFP transduced hASCs. In conclusion, our study demonstrates that adenovirus-mediated hTGF beta2 gene transfer is able to induce the hASCs into chondrogenic lineage both in vitro and in vivo. Ad5-hTGF beta2 transduced hASCs combined with three-dimensional PLGA/alginate compound may be a viable method in treating injuries of cartilage.

Ectopic neocartilage formation from predifferentiated human adipose derived stem cells induced by adenoviral-mediated transfer of hTGF beta2.

Chondrogenic potential of human adipose derived stem cells (hASCs) makes them a possible source of seeding cells for cartilage tissue engineering. In this study, chondrogenic differentiation of hASCs induced by transduction with replication-deficient adenovirus carrying human transforming growth factor beta2 (Ad5-hTGF beta2) was demonstrated by RT-PCR, immunohistochemistry staining, biochemical and western blot analysis. To evaluate if the in vitro differentiated hASCs could keep their chondrocytic phenotype and produce neo-cartilage in vivo, predifferentiated hASCs were seeded in different scaffolds and implanted in subcutaneous pockets on the dorsum of nude mice. After 4 and 12 weeks culture in vivo, specimens were harvested and examined by histological and immunohistochemical analysis, cartilage-like tissue formation was only found in alginate gel and PLGA/alginate compound groups, in PLGA group, fibrous tissues and angiogenesis ingrowth were observed. These findings demonstrated that adenovirus-mediated hTGF beta2 gene transfer could induce hASCs into a chondrogenic lineage in vitro, however, this predifferentiation did not guarantee ectopic cartilage formation in vivo unless appropriate three-dimensional scaffolds were used as the cell carry vehicles.

Neocartilage formation from predifferentiated human adipose derived stem cells in vivo.

AIM: To examine the chondrogenic potential of human adipose derived stem cells (hASC) induced by human transforming growth factor beta2 (hTGF beta2) in vitro, and to investigate if predifferentiated hASC can produce neocartilage in vivo. METHODS: hASC were isolated from subcutaneous adipose tissue and cultured in pellets with the addition of hTGF beta2. Chondrogenic differentiation was assayed by RT-PCR, Western blotting, toluidine blue staining, and immunohistochemistry staining for collagen type II. For the in vivo study, intact induced cell pellets or the released cells embedded in alginate gel with different concentrations were implanted subcutaneously in nude mice. Specimens were harvested at different time points and carried with histological and immunohistochemistry examination to evaluate the cartilage formation. RESULTS: RT-PCR analysis revealed that hASC produced aggrecan and collagen type II after 7 d of induction and continued throughout the culture period. This was also demonstrated by the Western blot analysis, positive staining of toluidine blue, and immunohistochemistry for collagen type II. After reseeding in the monolayer, the cells isolated from the pellets displayed a polygonal morphology compared with the primary spindle shape. hASC were released from the induced cell pellets when embedded in alginate gel (implanted cell concentration=5X10(6) /mL or higher). They produced neocartilage after 12 weeks in vivo culture; however, intact induced cell pellets implanted subcutaneously rapidly lost their differentiated phenotype. CONCLUSION: Chondrogenesis of hASC in vitro can be induced by combining pellet culture and hTGF beta2 treatment. Predifferentiated hASC embedded in alginate gel have the ability of producing neocartilage in vivo.

[Effects on VEGF and VEGF mRNA expression in issues of callus in treating fracture of rabbit with three-period treatment theory of bone fracture in traditional Chinese medicine].

OBJECTIVE: To observe the effects of the three-period treatment theory of bone fracture in traditional Chinese medicine (TCM) on VEGF and VEGF mRNA expression in the issues of callus of rabbits. And to explore the rationality of phasing method in TCM in treating fracture. METHOD: one hundred and forty male and healthy rabbits were made 3 mm wide bone defection at lower one third part of both radius as fracture healing model. Then those rabbits were divided into four groups randomly, which are three-period treatment group (TTG), one-period treatment group (OTG), positive medicine treatment group (PTG) and model control group (MCG). Those rabbits in TTG were treated with three-period treatment. Those in OTG were treated with one-period treatment. Those in PTG were feed by guzhecuoshangsan, a Chinese patent medicine which is used to treat bone fracture. Those in model control group were given no prescription or drug but distilled water as same dose as that of other groups. At day 3, 6, 9, 14, 28, 42 and 56, five rabbits were selected from every group randomly and were killed by aeroembolism respectively. Their radius were taken out and the left one was taken as the research object. Immunohistochemistry stain and in situ hybridization stain were performed to examinate the VEGF and VEGF mRNA expression in the haematoma, fibrous callus and soft callus. RESULT: All TCM treatment groups can enhance the VEGF and VEGF mRNA expression in the haematoma, fibrous callus and soft callus at different time points in fracture healing. The VEGF and VEGF mRNA expression in the three issues of TTG had the tendency of higher than that of the other groups at the most time points after operation. CONCLUSION: TCM can promote the VEGF and VEGF mRNA expression in the haematoma, fibrous callus and soft callus. Different Chinese medicines play various roles on VEGF and VEGF mRNA expression at different stage of fracture healing. Treating fracture in three-period treatment has more predominant effect on the expression of VEGF and VEGF mRNA in the haematoma, fibrous callus and soft callus than that of treating fracture with single prescription or drug. It is necessary to treat fracture in stages.

[Influence of nitrogen source NH4 Cl concentration on curdlan production in Alcaligenes faecalis].

The effect of initial ammonium chloride level on production of curdlan in Alcaligenesfaecalis was investigated. It was found that ammonium chloride was the limiting substrate for cell growth during the batch fermentation process. However, the cell growth and curdlan production could not be enhanced by solely increasing the initial ammonium chloride level. The pH drop in the broth due to the consumption of ammonium chloride also effected the cell growth and curdlan production. By simultaneously increasing the initial ammonium chloride concentration and implementing an optimal pH control strategy, which is to control pH at 7.0 in the growth phase, and then shift to 5.6 in the production phase, the biomass and curdlan production in batch fermentation were increased markedly. If the initial ammonium chloride concentration was increased from 1.1 g/L to 3.6 g/L, biomass concentration of 7.2 g/L was obtained, and the final curdlan concentration reached 30.5 g/L, which was 51.7% higher than that of the former case. As the cell growth was improved due to the increase of the initial ammonium chloride concentration, the agitation speed and aeration rates must be enhanced to suit the higher oxygen uptake requirement. However, as curdlan molecules is subject to the structural breakage due to the high shear stress at higher agitation speed, an overall optimal condition for both productivity and quality of curdlan should be considered comprehensively.