Surgical treatment of coronal shear fractures of the distal humerus with an intact lateral epicondyle by the lateral combined approach.

BACKGROUND: Coronal shear fractures of the distal humerus are not only rare and prone to misdiagnosis, but their surgical treatment can be challenging. We aimed to investigate the feasibility of exposing distal humeral coronal shear fractures with a combined lateral approach that preserves the extensors and lateral ulnar collateral ligament (LUCL) and to analyze the clinical efficacy of open reduction and internal fixation (ORIF) in the treatment of these injuries. METHODS: We included 45 patients who sustained coronal shear fractures of the distal humerus with the lateral epicondyle intact and were treated with ORIF from January 2013 to August 2020. The fractures were exposed by the lateral combined approach in which the tendons involving the common extensor, the extensor carpi ulnaris (ECU), and the LUCL were preserved. Two observation windows were formed anterior to and posterior to these tendons and the LUCL was used to achieve fracture reduction. Countersunk screws, with or without a plate placed on the posterior lateral condyle, were used to fix the fragments. The functional outcomes of these patients were reviewed and assessed with physical and radiographic examinations, range of motion (ROM) measurements, and self-evaluation Mayo Elbow Performance Index (MEPI) and Disabilities of the Arm, Shoulder, and Hand (DASH) scores. RESULTS: In total, 40 patients were followed up with for over 1 year and were included in the final analysis. The mean follow-up duration was 42±30 months (range, 12-107 months). The patients' mean age was 42 years (range, 14-74 years). According to the Dubberley Classification, there were 15 type I, 17 type II, and 8 type III fractures. At the final follow-up, the mean flexion-extension arc was 131° (range, 65-150) and mean pronation and supination was 73° (range, 45-80) and 71° (range, 40-80), respectively. The mean MEPI score was 88 (range, 61-97) points; the results were excellent in 21, good in 13, fair in 4, and poor in 2 patients. The mean DASH score was 11 (range, 0-42) points. Neither functional score nor range of movement was associated with age, sex, fracture type, injury type, or surgical timing. CONCLUSION: Reduction and stable fixation with internal fixation for coronal shear fractures of the distal humerus can be achieved by the lateral combined approach. Early functional mobilization allows for satisfactory restoration of elbow function.

Arthroscopic Medial Bi-portal Extra-articular Debridement for Recalcitrant Medial Epicondylitis.

Medial epicondylitis, or golfer's elbow, is characterized by pain and tenderness at the tendon insertion points of the pronator teres and flexor carpi radialis. Conservative treatment is sufficient for most patients, whereas surgical treatment is the best choice for intractable medial epicondylitis. With open surgery or arthroscopic surgery, good clinical results have been reported. However, there is still no consensus on which surgical technique is more ideal. We describe our technique of arthroscopic medial bi-portal extra-articular debridement, which is a safe and effective technique that allows more accurate debridement and maximum protection of the ulnar nerve while reducing surgical scars, relieving postoperative pain, reducing the probability of elbow infection and ankylosis, and shortening the recovery time.

Catalytic performance and mechanism study of the isomerization of 2,5-dichlorotoluene to 2,4-dichlorotoluene.

This work investigates the influence of catalyst HZSM-5 on the isomerization of 2,5-dichlorotoluene (2,5-DCT) to produce 2,4-dichlorotoluene (2,4-DCT). We observe that hydrothermal treatment leads to a decrease in total acidity and Brønsted/Lewis ratio of HZSM-5 while generating new secondary pores. These characteristics result in excellent selectivity for post-hydrothermal modified HZSM-5 in the isomerization reaction from 2,5-DCT to 2,4-DCT. Under atmospheric pressure at 350 °C, unmodified HZSM-5 achieves a selectivity of 66.4% for producing 2,4-DCT, however after hydrothermal modification the selectivity increases to 78.7%. Density Functional Theory (DFT) calculations explore the thermodynamic aspects of adsorption between the HZSM-5 surface and 2,4-DCT. The kinetic perspective investigates the mechanism involving proton attack on the methyl group of 2,5-DCT followed by rearrangement leading to formation of 2,4-DCT during isomerization. The consistency between simulation and experimental results provides evidence for the feasibility of isomerizing 2,5-DCT to 2,4-DCT. This work fills the gap in the low value-added product 2,5-DCT isomer conversion, indicating its significant practical application potential and provides a valuable reference and guidelines for industrial research in this field.

Epidemiological Characteristics of Major Joints Fracture-Dislocations.

OBJECTIVE: To describe the epidemiological features of major joints fracture-dislocations between 2015 and 2019. METHODS: This retrospective study enrolled patients with majorintra-articular fracture-dislocations who were treated in the third hospital of Hebei Medical University from January 2015 to December 2019. A total of 582 patients (389 [66.84%] males and 193 [33.16%] females) were identified. The distribution characteristics of intra-articular fracture-dislocations involving shoulder, elbow, wrist, hip, knee, and ankle joints were included. The potential associations between fractures with concomitant dislocations and related factors, such as age, gender and sites were explored. RESULTS: There were 92 cases (15.81%) of shoulder joints, 67 cases (11.51%) of elbow joints, 45 cases (7.73%) of wrist joints, 181 cases (31.10%) of hip joints, 42 cases (7.22%) of knee joints, and 155 cases (26.63%) of ankle joints. The overall male-to-female ratio was 2.02:1.The highest proportion age group of the six types intra-articular fracture-dislocations included the ages 25-34 years. For males, the highest proportion age group was 25-34 years, for females, it was 45-54 years. For male patients, hip was the most common, accounted for 35.48%, but ankle fracture-dislocation was the most common for females, accounted for 30.57%. The highest proportion age group of shoulder fracture-dislocation included the ages 55-64 years(22.83%), with a male to female ratio of 1.24:1. While the age group with the highest risk of elbow, wrist, hip, knee and ankle fracture- dislocation was 25-34 years (28.36%) with a male to female ratio of 2.19:1, 25-34 years (31.11%) with a male to female ratio of 8:1, 45-54 years (27.07%) with a male to female ratio of 3.21:1, 15-24 years (45.24%) with a male to female ratio of 0.75:1, 25-44 years (43.87%) with a male to female ratio of 1.63:1, respectively. The most common site of joint fracture-dislocation in different age groups was corresponding as follows, 0-14 years(elbow), 15-24 years(knee), 25-34 years(hip), 35-44 years(hip), 45-54 years(hip), 55-64 years(ankle), 65-74 years(shoulder), ≥75 years(shoulder). CONCLUSION: Major joints fracture-dislocations were most common in the hip and the least common in the knee, and there were more men than women. Hip was the most common affected joint in men while ankle in women. Age and sex factors can significantly affect the location of intra articular fracture and dislocation. The current study could aid orthopaedic surgeons in a better understanding of this injury and help to implement targeted preventive measures.

Age- and Gender-Specific Epidemiologic Characteristics of Major Intra-Articular Fractures: Five-Year Data from a Level 1 Trauma Center.

OBJECTIVE: To investigate the epidemiological characteristics of major intra-articular fractures. METHODS: This retrospective study enrolled patients with major intra-articular fractures who were treated in the Third Hospital of Hebei Medical University from January 2015 to December 2019. A total of 11,084 patients (7,338 [66.20%] males and 3,746 [33.80%] females) meeting the inclusion and exclusion criteria were included. The distribution characteristics of intra-articular fractures involving shoulder, elbow, wrist, hip, knee, ankle, and subtalar joints were identified.The potential associations between fractures and various other factors, such as age, gender, sites, were explored. RESULTS: There were 74 cases (0.67%) of shoulder fractures, 1,941 cases (17.51%) of elbow fractures, 1,155 cases (10.42%) of wrist fractures, 520 cases (4.69%) of hip fractures, 3,118 cases (28.13%) of knee fractures, 2,156 cases (19.45%) of ankle fractures, and 2,120 cases (19.13%) of subtalar fractures. The overall male-to-female ratio was 1.96:1. The highest proportion age group of major intra-articular fractures included the ages 45-54 years. For males, the highest proportion age group was 45-54 years, for females, it was 55-64 years. The knee joint fracture was the most common type, accounting for 28.13%. For male and female patients, knee fractures accounted for 26.19% and 31.93%, respectively, with a male to female ratio of 1.13:1. The proportion of shoulder fractures was the smallest among this investigation, accounting for 0.67%. For male and female patients, shoulder fractures accounted for 0.44% and 1.12%, respectively, with a male to female ratio of 0.76:1. The age group with the highest proportion of shoulder joint fractures was ≥65 year olds (41.89%), with a male to female ratio of 0.76:1. The age group with the highest risk of elbow, wrist, hip, knee, ankle, and subtalar joint fracture was 5-14 year olds (33.59%) with a male to female ratio of 3.29:1, 5-14 year olds (23.98%) with a male to female ratio of 6.91:1, 45-54 year olds (26.92%) with a male to female ratio of 5.67:1, 45-54 year olds (24.60%) with a male to female ratio of 1.68:1, 25-34 year olds (20.36%) with a male to female ratio of 2.30:1, 45-54 year olds (27.41%) with a male to female ratio of 9.02:1, respectively. The most common site of intra-articular fractures in different age groups was corresponding as follows: 0-4 year olds (elbow), 5-14 year olds (elbow), 15-24 year olds (ankle), 25-34 year olds (subtalar joint), 35-44 year olds (subtalar joint), 45-54 year olds (knee), 55-64 year olds (knee), 65-74 year olds (knee), and ≥75 year olds (knee). CONCLUSION: The current study revealed the age- and gender-specific epidemiological characteristics of major intra-articular fractures, providing a basis for clinical evaluation and practices.

Acupuncture for musculoskeletal pain: A meta-analysis and meta-regression of sham-controlled randomized clinical trials.

The aims of this systematic review were to study the analgesic effect of real acupuncture and to explore whether sham acupuncture (SA) type is related to the estimated effect of real acupuncture for musculoskeletal pain. Five databases were searched. The outcome was pain or disability immediately (≤1 week) following an intervention. Standardized mean differences (SMDs) with 95% confidence intervals were calculated. Meta-regression was used to explore possible sources of heterogeneity. Sixty-three studies (6382 individuals) were included. Eight condition types were included. The pooled effect size was moderate for pain relief (59 trials, 4980 individuals, SMD -0.61, 95% CI -0.76 to -0.47; P < 0.001) and large for disability improvement (31 trials, 4876 individuals, -0.77, -1.05 to -0.49; P < 0.001). In a univariate meta-regression model, sham needle location and/or depth could explain most or all heterogeneities for some conditions (e.g., shoulder pain, low back pain, osteoarthritis, myofascial pain, and fibromyalgia); however, the interactions between subgroups via these covariates were not significant (P < 0.05). Our review provided low-quality evidence that real acupuncture has a moderate effect (approximate 12-point reduction on the 100-mm visual analogue scale) on musculoskeletal pain. SA type did not appear to be related to the estimated effect of real acupuncture.

[Clinical observation of characteristic and treatment of posterior Pilon fractures].

OBJECTIVE: To explore clinical characteristics and treatment of posterior Pilon fracture. METHODS: From January 2011 to January 2013,18 patients with posterior Pilon fracures were treated. Among them, 13 were male and 5 were female, aged from 22 to 63 years old, with an average age of 46. All the patients were closed fractures. Open reduction and internal fixation were performed after swelling subsided, lateral malleolous and posterior Pilon fracture were exposured through lateral approach on healthy side, plates were used to fixed,screws or small plates were used to fix the posterior prominence of medial malleolus after changed to supine position. AOFAS scoring were applied to evaulate clinical effects. RESULTS: All patients were followed up with an average of 22(ranged, 12 to 48)months. All patients obtained satisfactory reset except one patient. All factures were recovered well with an average healing of 11 weeks. According to AOFAS score at the final following up, 7 cases were excellent,2 cases were moderate, and the total score was 86.8±9.2. CONCLUSION: Posterior Pilon fracture is not rare in clinical, its mechanism of injury, traumatic anatomy, surgical procedure and prognosis are different from that of classical ankle fracture and Pilon fracture.

[Clinical characteristics of triceps brachii tendon rupture at olecranon ending].

OBJECTIVE: To observe the clinical characteristics of triceps brachii tendon rupture at olecranon ending. METHODS: From June 2005 to November 2011,19 patients with triceps brachii tendon rupture at olecranon ending were treated with surgical technique. Among the 19 patients, 7 patients were male, with an average age of 24.1 years old (ranged, 15 to 41 years old); 12 patients were female, with an average age of 51.4 years old (ranged, 16 to 73 years old). Eight patients had injuries in the left elbows, and 11 patients had injuries in right elbows. Seventeen patients had injuries induced by walking fall and 2 patients had injuries induced by falling down. Thirteen patients were simple triceps brachii tendon rupture at olecranon ending, 6 patients were associated with other elbow injuries. Five patients were associated with radial fracture; 1 patient with capitellum fracture; 1 patient with coronoid process fracture; 1 patient with epitrochlear. All the lateral radiographs of the injuried elbow demenstrated the flecks of avulsed osseous material from the olecranon (flake sign). The associated injuries had the homologus presence. All the patients were treated with surgical techniques:15 patients were treated with figure-of-eight tension-band wire; figure-of-eight tension band wire and Kirschner wire in 1 patient; wire cerclage in 1 patient; nonabsorbable suture in 2 patients. The associated injuries were treated simultaneously. Plaster was applied after operation in 2 patients with heavier elbow associated injuries, other patients without any external fixation. The Mayo elbow score were observed to determinate the function of the elbow. RESULTS: All the patients were followed up, 1 patient died of other disease at one year after operation, the other 18 patients were followed up with an average of 47.9 months (ranged from 14 to 91 months). According to the Mayo elbow score, 16 patients got an excellent result and 2 good. CONCLUSION: Traumatic rupture of triceps brachii tendon at olecranon ending is not a rare injury, which is common in female older than fifty and in male younger then thirty. Surgical results are generally excellent. But dysfunction frequently remains in patients with associated elbow injuries.

Highly enantioselective oxidation of phenyl methyl sulfide and its derivatives into optically pure (S)-sulfoxides with Rhodococcus sp. CCZU10-1 in an n-octane-water biphasic system.

Enantiopure sulfoxides can be prepared via the asymmetric oxidation of sulfides using sulfide monooxygenases. The n-octane-water biphasic system was chosen for the bio-oxidation of a water-insoluble phenyl methyl sulfide (PMS) by Rhodococcus sp. CCZU10-1. In this n-octane-water system, the optimum reaction conditions were obtained. (S)-phenyl methyl sulfoxide ((S)-PMSO) with >99.9 % enantiomeric excess formed at 55.3 mM in the n-octane-water biphasic system. Using fed-batch method, a total of 118 mM (S)-PMSO accumulated in 1-L reaction mixture after the 7th feed, and no (R)-PMSO and sulfone were detected. Moreover, Rhodococcus sp. CCZU10-1 displayed fairly good activity and enantioselectivity toward other sulfides. In conclusion, Rhodococcus sp. CCZU10-1 is a promising biocatalyst for synthesizing highly optically active sulfoxides.

Improvement of drought and salt tolerance in Arabidopsis and Lotus corniculatus by overexpression of a novel DREB transcription factor from Populus euphratica.

A novel DREB (dehydration-responsive element binding) gene, designated PeDREB2a, was isolated from the desert-grown tree, Populus euphratica Oliv. PeDREB2a is classified into the A-5 group of DREB subfamily based on multiple sequence alignment and phylogenetic characterization. Using semi-quantitative RT-PCR, we found that the PeDREB2a was greatly induced by drought, NaCl, low temperature, 1-naphthaleneacetic acid (NAA), 6-benzyl aminopurine (6-BA) and gibberellic acid (GA3) treatments in P. euphratica seedling. Yeast transactivity assay demonstrated that PeDREB2a gene encodes a transcription activator. Overexpression of PeDREB2a under the stress-inducible rd29A promotor in transgenic Arabidopsis and Lotus corniculatus forage plants resulted in enhanced tolerance to salt and drought stresses. The PeDREB2a overexpressing Arabidopsis lines showed higher root length and plant height and had elevated levels of soluble sugars and lower levels of malondialdehyde under stress conditions compared to control plants. The results revealed that PeDREB2a play an essential role as a DREB transcription factor in regulation of stress-responsive signaling in P. euphratica.

[Typing and identification of non-polio enterovirus from acute flaccid paralysis cases in Ningxia, 1997-2011].

OBJECTIVE: To identify the serotype of 73 non-polio enterovirus (NPEV) strains from acute flaccid paralysis (AFP) cases in Ningxia province, during 1997 - 2011. METHODS: Partial sequencing of the VP1 region was amplified by RT-PCR with degenerate primers and sequenced while sequences were compared with the database of GenBank by the BLAST algorithm. Evolution was analyzed by constructing phylogenetic tree using Mega 5.1. RESULTS: In this study, a total of 73 NPEVs were analyzed, including 4 strains un-typed, 69 strains typed by RT-PCR. A total of 27 serotypes were identified, including 8 serotypes of human enterovirus (HEV)-A, 19 serotypes of HEV-B. The HEV-B group (46/69, 66.7%) constituted the largest proportion of isolates, followed by HEV-A (23/69, 33.3%), but no strains were found that belonged to HEV-C or HEV-D group. In the 69 strains, enterovirus 71 was the most frequently seen isolates, followed by coxsackie-virus A4, 16, 9 and echovirus 24, 6. CONCLUSION: HEV-B was the most predominant (46/69, 66.7%) serotype of NPEV in Ningxia during the AFP surveillance, in 1997 - 2011.

[Genetic characterization of coxsackievirus A16 isolated in Ningxia Hui Municipality in 2008].

OBJECTIVE: To study the genetic characterization of coxsackievirus A16 (CVA16) strains isolated during an epidemic of hand, foot, and mouth disease (HFMD) in Ningxia Hui Municipality in 2008. METHODS: Clinical samples were collected from HFMD patients in Ningxia Hui Municipality and CVA16 strains were isolated by viral isolation methods. Reverse transcription-polymerase chain reactions (RT-PCR), specific for CVA16 were performed with these CVA16 strains. Entire VP1 coding region amplification and sequencing were then performed and finally phylogenetic tree was constructed among Ningxia CVA16 strains and CVA16 representative strains of known genotypes and subgenotypes. RESULTS: 70 Ningxia CVA16 strains were isolated from HFMD patients in Ningxia in 2008 and the homology of nucleotide and amino acid were 90.8%-100.0% and 98.9% - 100.0%, respectively. Phylogenetic characteristics of the strains reconfirmed that they could be divided into two distinct genotypes-A and B. Genotype B could be further divided into the subgenotypes B1 and B2, while all the 70 Ningxia CVA16 strains belonged to the co-circulated clusters B1a and B1b within subgenotype B1, which belonged to 2 viral transmission chains. CONCLUSION: Our results indicated that subgenotype B1 CVA16 strains continued to circulate over a wide geographic area of mainland China since the first reported episode in Shenzhen city in 1999. Like other CVA16 strains isolated elsewhere in China, both B1a and B1b evolution branches were co-circulating in Ningxia Hui Municipality. Based on the close phylgenetic and chronological relationship with CVA16 isolated in other countries and regions near China. Our data confirmed that these strains co-evolved and co-circulated with those from neighboring countries and regions.

[Genetic characteristics of entervirus 71 isolated in Ningxia Hui Autonomous region in 2009].

OBJECTIVE: To analyze the genetic characteristics of EV71 strains isolated from HFMD cases in Ningxia Hui Autonomous Region in 2009. METHODS: In 2009, totally 385 specimens from 344 HFMD cases were collected from Ningxia. Enterovirus isolation was performed in RD cell line from all the specimens. EV71 isolates were identified by specific RT-PCR from the positive cultures, and sequences of complete EV71 VP1 encoding region were determined for farther analyses. RESULTS: Totally from 126 EV strains isolated in this study, 58 EV71 strains (46%) were identified. And complete VP1 sequences of 46 EV71 strains were determined, and genetic analyses were performed. It was showed that the nucleotide identity of 46 Ningxia strains with the representatives of A and B genotypes were 81.7%-82.8% and 83.1%-85.2%, and the amino acid identity were 93.9%-95.9% and 96. 2%-97.9% respectively. The nucleotide identity of NingXia EV71 isolates with representatives of subgenotype C1, C2, C3, C4a, C4b, and C5 were 88.3%-90.6% (97.9%-99.6%), 88.3%-90.1% (97.9%-99.3%), 87.8%-89.0% (97.6%-98.9%), 94.2%-98.9% (97.9%-100%), 91.8%-94.1% (98.6%-99.6%), and 86.7%-89.1% (97.9%-98.9%). Phylogenetic tree analysis revealed that 46 stains were clustered with reference stains of subgenotype C4 and the Ningxia EV71 isolates were belonged to subgenotype C4a. CONCLUSION: EV71 of subgenotype C4a had spread widely in Ningxia in 2009, which was absolutely predominant type in Ningxia in 2009 and also as the predominant type in China mainland since 2005.

[Genetic characterization of enterovirus 71 stains isolated in Ningxia province in 2008].

OBJECTIVE: To study the genetic characterization of enterovirus 71 (EV71) strains isolated from specimens of patients with hand-foot-mouth disease (HFMD) in Ningxia province in 2008. METHODS: All the stool, throat swab and vesicle samples that collected from patients with HFMD were cultured. The positive isolates were identified by reverse transcriptase PCR (RT-PCR) with specific primers of EV71. Complete VP1 gene sequences (891 nucleotides) of 29 strains (part of 93 EV71 strains) were determined and compared with A, B and C genotype reference EV71 strains while EV71 China isolates by homogeneity and phylogenetic tree analyses. RESULTS: 215 strains of EV were isolated from 439 specimens. Results from RT-PCR indicated that 93 strains belong to EV71. Phylogenetic tree analysis revealed that the selected 29 stains were clustered with reference strains of C4 subgenotype. The nucleotide identity with C4 reference strains was 91.7%-99.4%. The amino acid homogeneity was 96.6%-100.0%. CONCLUSION: The recently identified EV71 strains in Ningxia province belonged to subgenotype C4 which resembled to most of the isolates in China.

[Clinical observation on percutaneous autologous bone marrow grafting for treatment of fracture nonunion].

OBJECTIVE: To observe the effectiveness of percutaneous bone marrow grafting for treatment of fractures nonunion. METHODS: From June 2001 to December 2007, 29 consecutive cases of fractures nonunion were treated with percutaneous autologous bone marrow grafting included 20 males and 9 females, ranging in age from 20 to 71 years, with an average of 40 years. All the cases were traumatic fractures involving 13 of tibia, 10 of femur, 3 of humerus, 2 of ulna, 1 of radius, 11 cases of them were open fractures. All the cases were performed internal or external fixation before marrow grafting, intramedullary pin in 15 cases, plate in 12 cases, external fixator in 2 cases. The time from injury to therapy were from 6 to 12 months, with an average of 8.5 months. The type of nonunion included atrophic in 26 cases,hypertrophic in 3 cases. All the cases were performed 3 times injection, the interval was 1 month. According to the different fracture, the amount of bone marrow was from 6 to 15 ml. RESULTS: All the 29 cases were followed-up for from 5 to 22 months with an average of 14 months. Four of them were not observed obvious callus after 3 months from the 3rd injection, judged unsuccessful therapy, changed to perform autologous bone grafting (3 of them re-internal fixation), the follow-up ended. The other 25 cases obtained bone union during 3 to 8 months with an average of 4.5 months, the follow-up ended at the time of internal fixation removal. CONCLUSION: Percutaneous autologous bone marrow grafting is an effective, easy and economic therapy for fracture nonunion. But stable internal or external fixation is the premise. Excessive bone defect, the gap more than 5 mm and mal-align requiring rectification is not appropriate for this therapy.

[Secreted expression of nonstructural protein gene 3ABC of foot-and-mouth disease virus in Sf9 cells and activity analysis].

Entire 3ABC sequence of FMDV containing a 6 x his tag coding sequence at the N-terminal was obtained through PCR amplification using a pair of specific primers, subcloned into shuttle plasmid of pMelBac-B with a melittin secretion signal sequence and finally constructed recombinant plasmid of pMel-3ABC. After co-transfected the recombinant plasmid and linearized Bac-N-Blue DNA into Sf9 insect cell under intermediary agent of the Cellfectin, the result showed that we have already acquired recombinant baculovirus by screen of plaque assay and identification of PCR. Though the recombinant baculovirus infecting the Sf9 cells again, experiments indicated that 3ABC gene could express in insect cells and the expressed protein was secreted in the supernatant of Sf9 cell culture possessing favourable biological activities detected by adopting two methods of SDS-PAGE and Western blot. The result verified that the protein could respond with sera derived from FMDV infected animals, but have no responsibility with sera derived from health animals and vaccinated animals detected by indirect ELISA using antigen of expressed protein after purification with Ni-NTA his bind resin. Therefore, this study has established a solid foundation for establishing an effective diagnosis method to discriminating the FMDV infected animals from vaccinated animals.